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Recent Articles in PLoS Medicine / Public Libary of Science

Senechal B, Elain G, Jeziorski E, Grondin V, Patey-Mariaud de Serre N, Jaubert F, Beldjord K, Lellouch A, Glorion C, Zerah M, Mary P, Barkaoui M, Emile JF, Boccon-Gibod L, Josset P, Debr� M, Fischer A, Donadieu J, Geissmann F
Expansion of regulatory T cells in patients with Langerhans cell histiocytosis.
PLoS Med. 2007 Aug 14;4(8):e253.
BACKGROUND: Langerhans cell histiocytosis (LCH) is a rare clonal granulomatous disease that affects mainly children. LCH can involve various tissues such as bone, skin, lung, bone marrow, lymph nodes, and the central nervous system, and is frequently responsible for functional sequelae. The pathophysiology of LCH is unclear, but the uncontrolled proliferation of Langerhans cells (LCs) is believed to be the primary event in the formation of granulomas. The present study was designed to further investigate the nature of proliferating cells and the immune mechanisms involved in the LCH granulomas. METHODS AND FINDINGS: Biopsies (n = 24) and/or blood samples (n = 25) from 40 patients aged 0.25 to 13 y (mean 7.8 y), were studied to identify cells that proliferate in blood and granulomas. We found that the proliferating index of LCs was low ( approximately 1.9%), and we did not observe expansion of a monocyte or dendritic cell compartment in patients. We found that LCH lesions were a site of active inflammation, tissue remodeling, and neo-angiogenesis, and the majority of proliferating cells were endothelial cells, fibroblasts, and polyclonal T lymphocytes. Within granulomas, interleukin 10 was abundant, LCs expressed the TNF receptor family member RANK, and CD4(+) CD25(high) FoxP3(high) regulatory T cells (T-regs) represented 20% of T cells, and were found in close contact with LCs. FoxP3(+) T-regs were also expanded compared to controls, in the blood of LCH patients with active disease, among whom seven out of seven tested exhibited an impaired skin delayed-type hypersensitivity response. In contrast, the number of blood T-regs were normal after remission of LCH. CONCLUSIONS: These findings indicate that LC accumulation in LCH results from survival rather than uncontrolled proliferation, and is associated with the expansion of T-regs. These data suggest that LCs may be involved in the expansion of T-regs in vivo, resulting in the failure of the host immune system to eliminate LCH cells. Thus T-regs could be a therapeutic target in LCH. [Abstract/Link to Full Text]

Bates I, McKew S, Sarkinfada F
Anaemia: a useful indicator of neglected disease burden and control.
PLoS Med. 2007 Aug 14;4(8):e231. [Abstract/Link to Full Text]

Swen JJ, Huizinga TW, Gelderblom H, de Vries EG, Assendelft WJ, Kirchheiner J, Guchelaar HJ
Translating pharmacogenomics: challenges on the road to the clinic.
PLoS Med. 2007 Aug 14;4(8):e209.
Pharmacogenomics is one of the first clinical applications of the postgenomic era. It promises personalized medicine rather than the established "one size fits all" approach to drugs and dosages. The expected reduction in trial and error should ultimately lead to more efficient and safer drug therapy. In recent years, commercially available pharmacogenomic tests have been approved by the Food and Drug Administration (FDA), but their application in patient care remains very limited. More generally, the implementation of pharmacogenomics in routine clinical practice presents significant challenges. This article presents specific clinical examples of such challenges and discusses how obstacles to implementation of pharmacogenomic testing can be addressed. [Abstract/Link to Full Text]

Ramjee G, Govinden R, Morar NS, Mbewu A
South Africa's experience of the closure of the cellulose sulphate microbicide trial.
PLoS Med. 2007 Jul;4(7):e235. [Abstract/Link to Full Text]

Brennan MJ, Fruth U, Milstien J, Tiernan R, de Andrade Nishioka S, Chocarro L
Development of new tuberculosis vaccines: a global perspective on regulatory issues.
PLoS Med. 2007 Aug 7;4(8):e252. [Abstract/Link to Full Text]

McAlister FA, van Diepen S, Padwal RS, Johnson JA, Majumdar SR
How evidence-based are the recommendations in evidence-based guidelines?
PLoS Med. 2007 Aug 7;4(8):e250.
BACKGROUND: Treatment recommendations for the same condition from different guideline bodies often disagree, even when the same randomized controlled trial (RCT) evidence is cited. Guideline appraisal tools focus on methodology and quality of reporting, but not on the nature of the supporting evidence. This study was done to evaluate the quality of the evidence (based on consideration of its internal validity, clinical relevance, and applicability) underlying therapy recommendations in evidence-based clinical practice guidelines. METHODS AND FINDINGS: A cross-sectional analysis of cardiovascular risk management recommendations was performed for three different conditions (diabetes mellitus, dyslipidemia, and hypertension) from three pan-national guideline panels (from the United States, Canada, and Europe). Of the 338 treatment recommendations in these nine guidelines, 231 (68%) cited RCT evidence but only 105 (45%) of these RCT-based recommendations were based on high-quality evidence. RCT-based evidence was downgraded most often because of reservations about the applicability of the RCT to the populations specified in the guideline recommendation (64/126 cases, 51%) or because the RCT reported surrogate outcomes (59/126 cases, 47%). CONCLUSIONS: The results of internally valid RCTs may not be applicable to the populations, interventions, or outcomes specified in a guideline recommendation and therefore should not always be assumed to provide high-quality evidence for therapy recommendations. [Abstract/Link to Full Text]

Olson DR, Heffernan RT, Paladini M, Konty K, Weiss D, Mostashari F
Monitoring the impact of influenza by age: emergency department fever and respiratory complaint surveillance in New York City.
PLoS Med. 2007 Aug 7;4(8):e247.
BACKGROUND: The importance of understanding age when estimating the impact of influenza on hospitalizations and deaths has been well described, yet existing surveillance systems have not made adequate use of age-specific data. Monitoring influenza-related morbidity using electronic health data may provide timely and detailed insight into the age-specific course, impact and epidemiology of seasonal drift and reassortment epidemic viruses. The purpose of this study was to evaluate the use of emergency department (ED) chief complaint data for measuring influenza-attributable morbidity by age and by predominant circulating virus. METHODS AND FINDINGS: We analyzed electronically reported ED fever and respiratory chief complaint and viral surveillance data in New York City (NYC) during the 2001-2002 through 2005-2006 influenza seasons, and inferred dominant circulating viruses from national surveillance reports. We estimated influenza-attributable impact as observed visits in excess of a model-predicted baseline during influenza periods, and epidemic timing by threshold and cross correlation. We found excess fever and respiratory ED visits occurred predominantly among school-aged children (8.5 excess ED visits per 1,000 children aged 5-17 y) with little or no impact on adults during the early-2002 B/Victoria-lineage epidemic; increased fever and respiratory ED visits among children younger than 5 y during respiratory syncytial virus-predominant periods preceding epidemic influenza; and excess ED visits across all ages during the 2003-2004 (9.2 excess visits per 1,000 population) and 2004-2005 (5.2 excess visits per 1,000 population) A/H3N2 Fujian-lineage epidemics, with the relative impact shifted within and between seasons from younger to older ages. During each influenza epidemic period in the study, ED visits were increased among school-aged children, and each epidemic peaked among school-aged children before other impacted age groups. CONCLUSIONS: Influenza-related morbidity in NYC was highly age- and strain-specific. The impact of reemerging B/Victoria-lineage influenza was focused primarily on school-aged children born since the virus was last widespread in the US, while epidemic A/Fujian-lineage influenza affected all age groups, consistent with a novel antigenic variant. The correspondence between predominant circulating viruses and excess ED visits, hospitalizations, and deaths shows that excess fever and respiratory ED visits provide a reliable surrogate measure of incident influenza-attributable morbidity. The highly age-specific impact of influenza by subtype and strain suggests that greater age detail be incorporated into ongoing surveillance. Influenza morbidity surveillance using electronic data currently available in many jurisdictions can provide timely and representative information about the age-specific epidemiology of circulating influenza viruses. [Abstract/Link to Full Text]

Montori VM, Breslin M, Maleska M, Weymiller AJ
Creating a conversation: insights from the development of a decision aid.
PLoS Med. 2007 Aug 7;4(8):e233. [Abstract/Link to Full Text]

Chen Y, Whetstone HC, Lin AC, Nadesan P, Wei Q, Poon R, Alman BA
Beta-catenin signaling plays a disparate role in different phases of fracture repair: implications for therapy to improve bone healing.
PLoS Med. 2007 Jul 31;4(7):e249.
BACKGROUND: Delayed fracture healing causes substantial disability and usually requires additional surgical treatments. Pharmacologic management to improve fracture repair would substantially improve patient outcome. The signaling pathways regulating bone healing are beginning to be unraveled, and they provide clues into pharmacologic management. The beta-catenin signaling pathway, which activates T cell factor (TCF)-dependent transcription, has emerged as a key regulator in embryonic skeletogenesis, positively regulating osteoblasts. However, its role in bone repair is unknown. The goal of this study was to explore the role of beta-catenin signaling in bone repair. METHODS AND FINDINGS: Western blot analysis showed significant up-regulation of beta-catenin during the bone healing process. Using a beta-Gal activity assay to observe activation during healing of tibia fractures in a transgenic mouse model expressing a TCF reporter, we found that beta-catenin-mediated, TCF-dependent transcription was activated in both bone and cartilage formation during fracture repair. Using reverse transcription-PCR, we observed that several WNT ligands were expressed during fracture repair. Treatment with DKK1 (an antagonist of WNT/beta-catenin pathway) inhibited beta-catenin signaling and the healing process, suggesting that WNT ligands regulate beta-catenin. Healing was significantly repressed in mice conditionally expressing either null or stabilized beta-catenin alleles induced by an adenovirus expressing Cre recombinase. Fracture repair was also inhibited in mice expressing osteoblast-specific beta-catenin null alleles. In stark contrast, there was dramatically enhanced bone healing in mice expressing an activated form of beta-catenin, whose expression was restricted to osteoblasts. Treating mice with lithium activated beta-catenin in the healing fracture, but healing was enhanced only when treatment was started subsequent to the fracture. CONCLUSIONS: These results demonstrate that beta-catenin functions differently at different stages of fracture repair. In early stages, precise regulation of beta-catenin is required for pluripotent mesenchymal cells to differentiate to either osteoblasts or chondrocytes. Once these undifferentiated cells have become committed to the osteoblast lineage, beta-catenin positively regulates osteoblasts. This is a different function for beta-catenin than has previously been reported during development. Activation of beta-catenin by lithium treatment has potential to improve fracture healing, but only when utilized in later phases of repair, after mesenchymal cells have become committed to the osteoblast lineage. [Abstract/Link to Full Text]

Wiersinga WJ, Wieland CW, Dessing MC, Chantratita N, Cheng AC, Limmathurotsakul D, Chierakul W, Leendertse M, Florquin S, de Vos AF, White N, Dondorp AM, Day NP, Peacock SJ, van der Poll T
Toll-like receptor 2 impairs host defense in gram-negative sepsis caused by Burkholderia pseudomallei (Melioidosis).
PLoS Med. 2007 Jul 31;4(7):e248.
BACKGROUND: Toll-like receptors (TLRs) are essential in host defense against pathogens by virtue of their capacity to detect microbes and initiate the immune response. TLR2 is seen as the most important receptor for gram-positive bacteria, while TLR4 is regarded as the gram-negative TLR. Melioidosis is a severe infection caused by the gram-negative bacterium, Burkholderia pseudomallei, that is endemic in Southeast Asia. We aimed to characterize the expression and function of TLRs in septic melioidosis. METHODS AND FINDINGS: Patient studies: 34 patients with melioidosis demonstrated increased expression of CD14, TLR1, TLR2, and TLR4 on the cell surfaces of monocytes and granulocytes, and increased CD14, TLR1, TLR2, TLR4, LY96 (also known as MD-2), TLR5, and TLR10 mRNA levels in purified monocytes and granulocytes when compared with healthy controls. In vitro experiments: Whole-blood and alveolar macrophages obtained from TLR2 and TLR4 knockout (KO) mice were less responsive to B. pseudomallei in vitro, whereas in the reverse experiment, transfection of HEK293 cells with either TLR2 or TLR4 rendered these cells responsive to this bacterium. In addition, the lipopolysaccharide (LPS) of B. pseudomallei signals through TLR2 and not through TLR4. Mouse studies: Surprisingly, TLR4 KO mice were indistinguishable from wild-type mice with respect to bacterial outgrowth and survival in experimentally induced melioidosis. In contrast, TLR2 KO mice displayed a markedly improved host defenses as reflected by a strong survival advantage together with decreased bacterial loads, reduced lung inflammation, and less distant-organ injury. CONCLUSIONS: Patients with melioidosis displayed an up-regulation of multiple TLRs in peripheral blood monocytes and granulocytes. Although both TLR2 and TLR4 contribute to cellular responsiveness to B. pseudomallei in vitro, TLR2 detects the LPS of B. pseudomallei, and only TLR2 impacts on the immune response of the intact host in vivo. Inhibition of TLR2 may be a novel treatment strategy in melioidosis. [Abstract/Link to Full Text]

Wilson K, Gardam M, McDougall C, Attaran A, Upshur R
WHO's response to global public health threats: XDR-TB.
PLoS Med. 2007 Jul 31;4(7):e246. [Abstract/Link to Full Text]

How is WHO responding to global public health threats?
PLoS Med. 2007 May;4(5):e197. [Abstract/Link to Full Text]

Cunha A
Transparent development of WHO rapid advice guidelines: a useful approach.
PLoS Med. 2007 Jul 31;4(7):e245. [Abstract/Link to Full Text]

Sch�nemann HJ, Hill SR, Kakad M, Vist GE, Bellamy R, Stockman L, Wisl�ff TF, Del Mar C, Hayden F, Uyeki TM, Farrar J, Yazdanpanah Y, Zucker H, Beigel J, Chotpitayasunondh T, Hien TT, Ozbay B, Sugaya N, Oxman AD
Transparent development of the WHO rapid advice guidelines.
PLoS Med. 2007 May;4(5):e119.
Emerging health problems require rapid advice. We describe the development and pilot testing of a systematic, transparent approach used by the World Health Organization (WHO) to develop rapid advice guidelines in response to requests from member states confronted with uncertainty about the pharmacological management of avian influenza A (H5N1) virus infection. We first searched for systematic reviews of randomized trials of treatment and prevention of seasonal influenza and for non-trial evidence on H5N1 infection, including case reports and animal and in vitro studies. A panel of clinical experts, clinicians with experience in treating patients with H5N1, influenza researchers, and methodologists was convened for a two-day meeting. Panel members reviewed the evidence prior to the meeting and agreed on the process. It took one month to put together a team to prepare the evidence profiles (i.e., summaries of the evidence on important clinical and policy questions), and it took the team only five weeks to prepare and revise the evidence profiles and to prepare draft guidelines prior to the panel meeting. A draft manuscript for publication was prepared within 10 days following the panel meeting. Strengths of the process include its transparency and the short amount of time used to prepare these WHO guidelines. The process could be improved by shortening the time required to commission evidence profiles. Further development is needed to facilitate stakeholder involvement, and evaluate and ensure the guideline's usefulness. [Abstract/Link to Full Text]

Davey M
Surfactant levels in congenital diaphragmatic hernia.
PLoS Med. 2007 Jul 31;4(7):e243. [Abstract/Link to Full Text]

Boucherat O, Benachi A, Chailley-Heu B, Franco-Montoya ML, Elie C, Martinovic J, Bourbon JR
Surfactant maturation is not delayed in human fetuses with diaphragmatic hernia.
PLoS Med. 2007 Jul 31;4(7):e237.
BACKGROUND: Pulmonary hypoplasia and persistent pulmonary hypertension account for significant mortality and morbidity in neonates with congenital diaphragmatic hernia (CDH). Global lung immaturity and studies in animal models suggest the presence of surfactant deficiency that may further complicate the pathophysiology of CDH. However, data about surfactant status in human fetuses with CDH at birth are contradictory. The lack of a chronological study of surfactant content in late pregnancy has been a significant limitation. The appropriateness of administering surfactant supplements to neonates with CDH is therefore a debated question. METHODS AND FINDINGS: We investigated surfactant content in human fetuses with CDH compared to age-matched fetuses with nonpulmonary diseases used as controls. Concentrations of disaturated phosphatidylcholine and surfactant proteins were found to be similar at a given stage of pregnancy, with both components showing a similar pattern of increase with progressing pregnancy in fetuses with CDH and in control fetuses. Thyroid transcription factor 1, a critical regulator of surfactant protein transcription, similarly displayed no difference in abundance. Finally, we examined the expression of three glucocorticoid-regulated diffusible mediators involved in lung epithelial maturation, namely: keratinocyte growth factor (KGF), leptin, and neuregulin 1 beta 1 (NRG1-beta1). KGF expression decreased slightly with time in control fetuses, but remained unchanged in fetuses with CDH. Leptin and NRG1-beta1 similarly increased in late pregnancy in control and CDH lungs. These maturation factors were also determined in the sheep fetus with surgical diaphragmatic hernia, in which surfactant deficiency has been reported previously. In contrast to the findings in humans, surgical diaphragmatic hernia in the sheep fetus was associated with decreased KGF and neuregulin expression. Fetoscopic endoluminal tracheal occlusion performed in the sheep model to correct lung hypoplasia increased leptin expression, partially restored KGF expression, and fully restored neuregulin expression. CONCLUSIONS: Our results indicate that CDH does not impair surfactant storage in human fetuses. CDH lungs exhibited no trend toward a decrease in contents, or a delay in developmental changes for any of the studied surfactant components and surfactant maturation factors. Surfactant amounts are likely to be appropriate to lung size. These findings therefore do not support the use of surfactant therapy for infants with CDH. Moreover, they raise the question of the relevance of CDH animal models to explore lung biochemical maturity. [Abstract/Link to Full Text]

Aponte JJ, Menendez C, Schellenberg D, Kahigwa E, Mshinda H, Vountasou P, Tanner M, Alonso PL
Age interactions in the development of naturally acquired immunity to Plasmodium falciparum and its clinical presentation.
PLoS Med. 2007 Jul 31;4(7):e242.
BACKGROUND: Naturally acquired malaria immunity has many determinants and, in the absence of immunological markers of protection, studies assessing malaria incidence through clinical endpoints remain an approach to defining immunity acquisition. We investigated the role of age in disease incidence and the effects of chemoprophylaxis on clinical immunity development to Plasmodium falciparum during a randomised controlled trial. METHODS AND FINDINGS: A total of 415 Tanzanian infants were randomly assigned to receive weekly malaria prophylaxis with Deltaprim (3.125 mg of pyrimethamine plus 25 mg of dapsone) or placebo between the ages of 2 and 12 mo. Children were followed up until 4 y of age. Uncomplicated febrile malaria, severe malaria, and anaemia morbidity were assessed through hospital-based passive surveillance. Compared with the group of control participants, there was a marked reduction in the incidence of clinical malaria, severe malaria, and anaemia in the group of children who had received chemoprophylaxis during the first year of life. After discontinuing the intervention, there was a significant increase in the incidence of clinical malaria for 2 y. The cumulative rates of clinical malaria, by age 4 y, were slightly higher in the group of children who had previously received chemoprophylaxis: 3.22 episodes versus 3.02 episodes in the group of control participants; rate difference 0.20 (95% confidence interval [CI]: -0.21 to 0.59). By age 4 y, the cumulative rates of severe malaria, however, were slightly lower in chemosuppressed children (0.47 versus 0.59) (rate difference -0.12 [95% CI: -0.27 to 0.03]). The number of episodes of anaemia was also slightly lower in chemosuppressed children by age 4 y: 0.93 episodes (95% CI: 0.79 to 0.97) versus 1.12 episodes in the group of control participants (95% CI: 0.97 to 1.28) (rate difference -0.19 [95% CI: -0.40 to 0.01]), respectively. CONCLUSIONS: Reducing exposure to P. falciparum antigens through chemoprophylaxis early in life can delay immunity acquisition. Infants appear to acquire immunity faster than older children, but have a higher risk of developing severe forms of malaria and anaemia. These findings provide insight on the interplay between immunity and exposure-reduction interventions. [Abstract/Link to Full Text]

de O�ate WA
Medicines without Doctors: in Mozambique, salaries are not the biggest problem.
PLoS Med. 2007 Jul 31;4(7):e236. [Abstract/Link to Full Text]

Ooms G, Van Damme W, Temmerman M
Medicines without doctors: why the Global Fund must fund salaries of health workers to expand AIDS treatment.
PLoS Med. 2007 Apr;4(4):e128. [Abstract/Link to Full Text]

Kishore SP, Dhadialla PS
A student-led campaign to help tackle neglected tropical diseases.
PLoS Med. 2007 Jul 24;4(7):e241. [Abstract/Link to Full Text]

Dahan A, Nieuwenhuijs D, Teppema L
Plasticity of central chemoreceptors: effect of bilateral carotid body resection on central CO2 sensitivity.
PLoS Med. 2007 Jul 24;4(7):e239.
BACKGROUND: Human breathing is regulated by feedback and feed-forward control mechanisms, allowing a strict matching between metabolic needs and the uptake of oxygen in the lungs. The most important control mechanism, the metabolic ventilatory control system, is fine-tuned by two sets of chemoreceptors, the peripheral chemoreceptors in the carotid bodies (located in the bifurcation of the common carotid arteries) and the central CO2 chemoreceptors in the ventral medulla. Animal data indicate that resection of the carotid bodies results, apart from the loss of the peripheral chemoreceptors, in reduced activity of the central CO2 sensors. We assessed the acute and chronic effect of carotid body resection in three humans who underwent bilateral carotid body resection (bCBR) after developing carotid body tumors. METHODS AND FINDINGS: The three patients (two men, one woman) were suffering from a hereditary form of carotid body tumors. They were studied prior to surgery and at regular intervals for 2-4 y following bCBR. We obtained inspired minute ventilation (Vi) responses to hypoxia and CO2. The Vi-CO2 responses were separated into a peripheral (fast) response and a central (slow) response with a two-compartment model of the ventilatory control system. Following surgery the ventilatory CO2 sensitivity of the peripheral chemoreceptors and the hypoxic responses were not different from zero or below 10% of preoperative values. The ventilatory CO2 sensitivity of the central chemoreceptors decreased by about 75% after surgery, with peak reduction occurring between 3 and 6 mo postoperatively. This was followed by a slow return to values close to preoperative values within 2 y. During this slow return, the Vi-CO2 response shifted slowly to the right by about 8 mm Hg. CONCLUSIONS: The reduction in central Vi-CO2 sensitivity after the loss of the carotid bodies suggests that the carotid bodies exert a tonic drive or tonic facilitation on the output of the central chemoreceptors that is lost upon their resection. The observed return of the central CO2 sensitivity is clear evidence for central plasticity within the ventilatory control system. Our data, although of limited sample size, indicate that the response mechanisms of the ventilatory control system are not static but depend on afferent input and exhibit a large degree of restoration or plasticity. In addition, the permanent absence of the breathing response to hypoxia after bCBR may aggravate the pathological consequences of sleep-disordered breathing. [Abstract/Link to Full Text]

Munro SA, Lewin SA, Smith HJ, Engel ME, Fretheim A, Volmink J
Patient adherence to tuberculosis treatment: a systematic review of qualitative research.
PLoS Med. 2007 Jul 24;4(7):e238.
BACKGROUND: Tuberculosis (TB) is a major contributor to the global burden of disease and has received considerable attention in recent years, particularly in low- and middle-income countries where it is closely associated with HIV/AIDS. Poor adherence to treatment is common despite various interventions aimed at improving treatment completion. Lack of a comprehensive and holistic understanding of barriers to and facilitators of, treatment adherence is currently a major obstacle to finding effective solutions. The aim of this systematic review of qualitative studies was to understand the factors considered important by patients, caregivers and health care providers in contributing to TB medication adherence. METHODS AND FINDINGS: We searched 19 electronic databases (1966-February 2005) for qualitative studies on patients', caregivers', or health care providers' perceptions of adherence to preventive or curative TB treatment with the free text terms "Tuberculosis AND (adherence OR compliance OR concordance)". We supplemented our search with citation searches and by consulting experts. For included studies, study quality was assessed using a predetermined checklist and data were extracted independently onto a standard form. We then followed Noblit and Hare's method of meta-ethnography to synthesize the findings, using both reciprocal translation and line-of-argument synthesis. We screened 7,814 citations and selected 44 articles that met the prespecified inclusion criteria. The synthesis offers an overview of qualitative evidence derived from these multiple international studies. We identified eight major themes across the studies: organisation of treatment and care; interpretations of illness and wellness; the financial burden of treatment; knowledge, attitudes, and beliefs about treatment; law and immigration; personal characteristics and adherence behaviour; side effects; and family, community, and household support. Our interpretation of the themes across all studies produced a line-of-argument synthesis describing how four major factors interact to affect adherence to TB treatment: structural factors, including poverty and gender discrimination; the social context; health service factors; and personal factors. The findings of this study are limited by the quality and foci of the included studies. CONCLUSIONS: Adherence to the long course of TB treatment is a complex, dynamic phenomenon with a wide range of factors impacting on treatment-taking behaviour. Patients' adherence to their medication regimens was influenced by the interaction of a number of these factors. The findings of our review could help inform the development of patient-centred interventions and of interventions to address structural barriers to treatment adherence. [Abstract/Link to Full Text]

Sullivan PS, Kilmarx PH, Peterman TA, Taylor AW, Nakashima AK, Kamb ML, Warner L, Mastro TD
Male circumcision for prevention of HIV transmission: what the new data mean for HIV prevention in the United States.
PLoS Med. 2007 Jul 24;4(7):e223. [Abstract/Link to Full Text]

Masenior NF, Beyrer C
The US anti-prostitution pledge: First Amendment challenges and public health priorities.
PLoS Med. 2007 Jul 24;4(7):e207. [Abstract/Link to Full Text]

Silverman J, Decker M
The US anti-prostitution pledge: a call for cooperation.
PLoS Med. 2007 Oct 30;4(10):1702; author reply 1702-3. [Abstract/Link to Full Text]

Fern�ndez Taylor KR
Excessive work hours of physicians in training in El Salvador: putting patients at risk.
PLoS Med. 2007 Jul;4(7):e205. [Abstract/Link to Full Text]

Kasi PM, Kassi M, Khawar T
Excessive work hours of physicians in training: maladaptive coping strategies.
PLoS Med. 2007 Sep 25;4(9):e279. [Abstract/Link to Full Text]

Fretheim A, H�velsrud K, MacLennan G, Kristoffersen DT, Oxman AD
The effects of mandatory prescribing of thiazides for newly treated, uncomplicated hypertension: interrupted time-series analysis.
PLoS Med. 2007 Jul;4(7):e232.
BACKGROUND: The purpose of our study was to evaluate the effects of a new reimbursement rule for antihypertensive medication that made thiazides mandatory first-line drugs for newly treated, uncomplicated hypertension. The objective of the new regulation was to reduce drug expenditures. METHODS AND FINDINGS: We conducted an interrupted time-series analysis on prescribing data before and after the new reimbursement rule for antihypertensive medication was put into effect. All patients started on antihypertensive medication in 61 general practices in Norway were included in the analysis. The new rule was put forward by the Ministry of Health and was approved by parliament. Adherence to the rule was monitored only minimally, and there were no penalties for non-adherence. Our primary outcome was the proportion of thiazide prescriptions among all prescriptions made for persons started on antihypertensive medication. Secondary outcomes included the proportion of patients who, within 4 mo, reached recommended blood-pressure goals and the proportion of patients who, within 4 mo, were not started on a second antihypertensive drug. We also compared drug costs before and after the intervention. During the baseline period, 10% of patients started on antihypertensive medication were given a thiazide prescription. This proportion rose steadily during the transition period, after which it remained stable at 25%. For other outcomes, no statistically significant differences were demonstrated. Achievement of treatment goals was slightly higher (56.6% versus 58.4%) after the new rule was introduced, and the prescribing of a second drug was slightly lower (24.0% versus 21.8%). Drug costs were reduced by an estimated Norwegian kroner 4.8 million (0.58 million Euros, US$0.72 million) in the first year, which is equivalent to Norwegian kroner 1.06 per inhabitant (0.13 Euros, US$0.16). CONCLUSIONS: Prescribing of thiazides in Norway for uncomplicated hypertension more than doubled after a reimbursement rule requiring the use of thiazides as the first-choice therapy was put into effect. However, the resulting savings on drug expenditures were modest. There were no significant changes in the achievement of treatment goals or in the prescribing of a second antihypertensive drug. [Abstract/Link to Full Text]

Sripa B, Kaewkes S, Sithithaworn P, Mairiang E, Laha T, Smout M, Pairojkul C, Bhudhisawasdi V, Tesana S, Thinkamrop B, Bethony JM, Loukas A, Brindley PJ
Liver fluke induces cholangiocarcinoma.
PLoS Med. 2007 Jul;4(7):e201. [Abstract/Link to Full Text]

Riddell SR, Appelbaum FR
Graft-versus-host disease: a surge of developments.
PLoS Med. 2007 Jul;4(7):e198. [Abstract/Link to Full Text]

Killeen GF, Smith TA, Ferguson HM, Mshinda H, Abdulla S, Lengeler C, Kachur SP
Preventing childhood malaria in Africa by protecting adults from mosquitoes with insecticide-treated nets.
PLoS Med. 2007 Jul;4(7):e229.
BACKGROUND: Malaria prevention in Africa merits particular attention as the world strives toward a better life for the poorest. Insecticide-treated nets (ITNs) represent a practical means to prevent malaria in Africa, so scaling up coverage to at least 80% of young children and pregnant women by 2010 is integral to the Millennium Development Goals (MDG). Targeting individual protection to vulnerable groups is an accepted priority, but community-level impacts of broader population coverage are largely ignored even though they may be just as important. We therefore estimated coverage thresholds for entire populations at which individual- and community-level protection are equivalent, representing rational targets for ITN coverage beyond vulnerable groups. METHODS AND FINDINGS: Using field-parameterized malaria transmission models, we show that high (80% use) but exclusively targeted coverage of young children and pregnant women (representing <20% of the population) will deliver limited protection and equity for these vulnerable groups. In contrast, relatively modest coverage (35%-65% use, with this threshold depending on ecological scenario and net quality) of all adults and children, rather than just vulnerable groups, can achieve equitable community-wide benefits equivalent to or greater than personal protection. CONCLUSIONS: Coverage of entire populations will be required to accomplish large reductions of the malaria burden in Africa. While coverage of vulnerable groups should still be prioritized, the equitable and communal benefits of wide-scale ITN use by older children and adults should be explicitly promoted and evaluated by national malaria control programmes. ITN use by the majority of entire populations could protect all children in such communities, even those not actually covered by achieving existing personal protection targets of the MDG, Roll Back Malaria Partnership, or the US President's Malaria Initiative. [Abstract/Link to Full Text]

Mathenge W, Nkurikiye J, Limburg H, Kuper H
Rapid assessment of avoidable blindness in Western Rwanda: blindness in a postconflict setting.
PLoS Med. 2007 Jul;4(7):e217.
BACKGROUND: The World Health Organization estimates that there were 37 million blind people in 2002 and that the prevalence of blindness was 9% among adults in Africa aged 50 years or older. Recent surveys indicate that this figure may be overestimated, while a survey from southern Sudan suggested that postconflict areas are particularly vulnerable to blindness. The aim of this study was to conduct a Rapid Assessment for Avoidable Blindness to estimate the magnitude and causes of visual impairment in people aged > or = 50 y in the postconflict area of the Western Province of Rwanda, which includes one-quarter of the population of Rwanda. METHODS AND FINDINGS: Clusters of 50 people aged > or = 50 y were selected through probability proportionate to size sampling. Households within clusters were selected through compact segment sampling. Visual acuity (VA) was measured with a tumbling "E" chart, and those with VA below 6/18 in either eye were examined by an ophthalmologist. The teams examined 2,206 people (response rate 98.0%). The unadjusted prevalence of bilateral blindness was 1.8% (95% confidence interval [CI] 1.2%-2.4%), 1.3% (0.8%-1.7%) for severe visual impairment, and 5.3% (4.2%-6.4%) for visual impairment. Most bilateral blindness (65%) was due to cataract. Overall, the vast majority of cases of blindness (80.0%), severe visual impairment (67.9%), and visual impairment (87.2%) were avoidable (i.e.. due to cataract, refractive error, aphakia, trachoma, or corneal scar). The cataract surgical coverage was moderate; 47% of people with bilateral cataract blindness (VA < 3/60) had undergone surgery. Of the 29 eyes that had undergone cataract surgery, nine (31%) had a best-corrected poor outcome (i.e., VA < 6/60). Extrapolating these estimates to Rwanda's Western Province, among the people aged 50 years or above 2,565 are expected to be blind, 1,824 to have severe visual impairment, and 8,055 to have visual impairment. CONCLUSIONS: The prevalence of blindness and visual impairment in this postconflict area in the Western Province of Rwanda was far lower than expected. Most of the cases of blindness and visual impairment remain avoidable, however, suggesting that the implementation of an effective eye care service could reduce the prevalence further. [Abstract/Link to Full Text]

Gartner CE, Hall WD, Chapman S, Freeman B
Should the health community promote smokeless tobacco (snus) as a harm reduction measure?
PLoS Med. 2007 Jul;4(7):e185.
BACKGROUND TO THE DEBATE: The tobacco control community is divided on whether or not to inform the public that using oral, smokeless tobacco (Swedish snus) is less hazardous to health than smoking tobacco. Proponents of "harm reduction" point to the Swedish experience. Snus seems to be widely used as an alternative to cigarettes in Sweden, say these proponents, contributing to the low overall prevalence of smoking and smoking-related disease. Harm reduction proponents thus argue that the health community should actively inform inveterate cigarette smokers of the benefits of switching to snus. However, critics of harm reduction say that snus has its own risks, that no form of tobacco should ever be promoted, and that Sweden's experience is likely to be specific to that culture and not transferable to other settings. Critics also remain deeply suspicious that the tobacco industry will use snus marketing as a "gateway" to promote cigarettes. In the interests of promoting debate, the authors (who are collaborators on a research project on the future of tobacco control) have agreed to outline the strongest arguments for and against promoting Swedish snus as a form of harm reduction. [Abstract/Link to Full Text]

Williamson J, Proctor C
Should the health community promote smokeless tobacco (snus): comments from British American Tobacco.
PLoS Med. 2007 Oct 30;4(10):1703-4; author reply 1704-5. [Abstract/Link to Full Text]

Aguirre GK, Kom�romy AM, Cideciyan AV, Brainard DH, Aleman TS, Roman AJ, Avants BB, Gee JC, Korczykowski M, Hauswirth WW, Acland GM, Aguirre GD, Jacobson SG
Canine and human visual cortex intact and responsive despite early retinal blindness from RPE65 mutation.
PLoS Med. 2007 Jun;4(6):e230.
BACKGROUND: RPE65 is an essential molecule in the retinoid-visual cycle, and RPE65 gene mutations cause the congenital human blindness known as Leber congenital amaurosis (LCA). Somatic gene therapy delivered to the retina of blind dogs with an RPE65 mutation dramatically restores retinal physiology and has sparked international interest in human treatment trials for this incurable disease. An unanswered question is how the visual cortex responds after prolonged sensory deprivation from retinal dysfunction. We therefore studied the cortex of RPE65-mutant dogs before and after retinal gene therapy. Then, we inquired whether there is visual pathway integrity and responsivity in adult humans with LCA due to RPE65 mutations (RPE65-LCA). METHODS AND FINDINGS: RPE65-mutant dogs were studied with fMRI. Prior to therapy, retinal and subcortical responses to light were markedly diminished, and there were minimal cortical responses within the primary visual areas of the lateral gyrus (activation amplitude mean +/- standard deviation [SD] = 0.07% +/- 0.06% and volume = 1.3 +/- 0.6 cm(3)). Following therapy, retinal and subcortical response restoration was accompanied by increased amplitude (0.18% +/- 0.06%) and volume (8.2 +/- 0.8 cm(3)) of activation within the lateral gyrus (p < 0.005 for both). Cortical recovery occurred rapidly (within a month of treatment) and was persistent (as long as 2.5 y after treatment). Recovery was present even when treatment was provided as late as 1-4 y of age. Human RPE65-LCA patients (ages 18-23 y) were studied with structural magnetic resonance imaging. Optic nerve diameter (3.2 +/- 0.5 mm) was within the normal range (3.2 +/- 0.3 mm), and occipital cortical white matter density as judged by voxel-based morphometry was slightly but significantly altered (1.3 SD below control average, p = 0.005). Functional magnetic resonance imaging in human RPE65-LCA patients revealed cortical responses with a markedly diminished activation volume (8.8 +/- 1.2 cm(3)) compared to controls (29.7 +/- 8.3 cm(3), p < 0.001) when stimulated with lower intensity light. Unexpectedly, cortical response volume (41.2 +/- 11.1 cm(3)) was comparable to normal (48.8 +/- 3.1 cm(3), p = 0.2) with higher intensity light stimulation. CONCLUSIONS: Visual cortical responses dramatically improve after retinal gene therapy in the canine model of RPE65-LCA. Human RPE65-LCA patients have preserved visual pathway anatomy and detectable cortical activation despite limited visual experience. Taken together, the results support the potential for human visual benefit from retinal therapies currently being aimed at restoring vision to the congenitally blind with genetic retinal disease. [Abstract/Link to Full Text]

Brogger J
Reporting of systematic reviews: better software required.
PLoS Med. 2007 Jun;4(6):e225. [Abstract/Link to Full Text]

Moher D, Tetzlaff J, Tricco AC, Sampson M, Altman DG
Epidemiology and reporting characteristics of systematic reviews.
PLoS Med. 2007 Mar 27;4(3):e78.
BACKGROUND: Systematic reviews (SRs) have become increasingly popular to a wide range of stakeholders. We set out to capture a representative cross-sectional sample of published SRs and examine them in terms of a broad range of epidemiological, descriptive, and reporting characteristics, including emerging aspects not previously examined. METHODS AND FINDINGS: We searched Medline for SRs indexed during November 2004 and written in English. Citations were screened and those meeting our inclusion criteria were retained. Data were collected using a 51-item data collection form designed to assess the epidemiological and reporting details and the bias-related aspects of the reviews. The data were analyzed descriptively. In total 300 SRs were identified, suggesting a current annual publication rate of about 2,500, involving more than 33,700 separate studies including one-third of a million participants. The majority (272 [90.7%]) of SRs were reported in specialty journals. Most reviews (213 [71.0%]) were categorized as therapeutic, and included a median of 16 studies involving 1,112 participants. Funding sources were not reported in more than one-third (122 [40.7%]) of the reviews. Reviews typically searched a median of three electronic databases and two other sources, although only about two-thirds (208 [69.3%]) of them reported the years searched. Most (197/295 [66.8%]) reviews reported information about quality assessment, while few (68/294 [23.1%]) reported assessing for publication bias. A little over half (161/300 [53.7%]) of the SRs reported combining their results statistically, of which most (147/161 [91.3%]) assessed for consistency across studies. Few (53 [17.7%]) SRs reported being updates of previously completed reviews. No review had a registration number. Only half (150 [50.0%]) of the reviews used the term "systematic review" or "meta-analysis" in the title or abstract. There were large differences between Cochrane reviews and non-Cochrane reviews in the quality of reporting several characteristics. CONCLUSIONS: SRs are now produced in large numbers, and our data suggest that the quality of their reporting is inconsistent. This situation might be improved if more widely agreed upon evidence-based reporting guidelines were endorsed and adhered to by authors and journals. These results substantiate the view that readers should not accept SRs uncritically. [Abstract/Link to Full Text]

Many reviews are systematic but some are more transparent and completely reported than others.
PLoS Med. 2007 Mar;4(3):e147. [Abstract/Link to Full Text]

The changing face of occupational medicine.
PLoS Med. 2007 Jun;4(6):e221. [Abstract/Link to Full Text]

Grudzen CR, Kerndt PR
The adult film industry: time to regulate?
PLoS Med. 2007 Jun;4(6):e126. [Abstract/Link to Full Text]

Villafuerte-G�lvez J, Curioso WH, Gayoso O
Biomedical journals and global poverty: is HINARI a step backwards?
PLoS Med. 2007 Jun;4(6):e220. [Abstract/Link to Full Text]

Ioannidis JP
Why most published research findings are false: author's reply to Goodman and Greenland.
PLoS Med. 2007 Jun;4(6):e215. [Abstract/Link to Full Text]

Goodman S, Greenland S
Why most published research findings are false: problems in the analysis.
PLoS Med. 2007 Apr;4(4):e168. [Abstract/Link to Full Text]

Reinhard R
Consent for genomic epidemiology in developing countries: added human subject protection also needed.
PLoS Med. 2007 Jun;4(6):e214. [Abstract/Link to Full Text]

Bonchek L
Lethal injection: let's be honest about the death penalty.
PLoS Med. 2007 Jun;4(6):e213; discussion e224. [Abstract/Link to Full Text]

Bermejo-Martin JF, Kelvin DJ, Guan Y, Chen H, Perez-Bre�a P, Casas I, Arranz E, de Lejarazu RO
Neuraminidase antibodies and H5N1: geographic-dependent influenza epidemiology could determine cross-protection against emerging strains.
PLoS Med. 2007 Jun;4(6):e212. [Abstract/Link to Full Text]

Sandbulte MR, Jimenez GS, Boon AC, Smith LR, Treanor JJ, Webby RJ
Cross-reactive neuraminidase antibodies afford partial protection against H5N1 in mice and are present in unexposed humans.
PLoS Med. 2007 Feb;4(2):e59.
BACKGROUND: A pandemic H5N1 influenza outbreak would be facilitated by an absence of immunity to the avian-derived virus in the human population. Although this condition is likely in regard to hemagglutinin-mediated immunity, the neuraminidase (NA) of H5N1 viruses (avN1) and of endemic human H1N1 viruses (huN1) are classified in the same serotype. We hypothesized that an immune response to huN1 could mediate cross-protection against H5N1 influenza virus infection. METHODS AND FINDINGS: Mice were immunized against the NA of a contemporary human H1N1 strain by DNA vaccination. They were challenged with recombinant A/Puerto Rico/8/34 (PR8) viruses bearing huN1 (PR8-huN1) or avN1 (PR8-avN1) or with H5N1 virus A/Vietnam/1203/04. Additional na�ve mice were injected with sera from vaccinated mice prior to H5N1 challenge. Also, serum specimens from humans were analyzed for reactivity with avN1. Immunization elicited a serum IgG response to huN1 and robust protection against the homologous challenge virus. Immunized mice were partially protected from lethal challenge with H5N1 virus or recombinant PR8-avN1. Sera transferred from immunized mice to na�ve animals conferred similar protection against H5N1 mortality. Analysis of human sera showed that antibodies able to inhibit the sialidase activity of avN1 exist in some individuals. CONCLUSIONS: These data reveal that humoral immunity elicited by huN1 can partially protect against H5N1 infection in a mammalian host. Our results suggest that a portion of the human population could have some degree of resistance to H5N1 influenza, with the possibility that this could be induced or enhanced through immunization with seasonal influenza vaccines. [Abstract/Link to Full Text]

Khoury MJ, Little J, Higgins J, Ioannidis JP, Gwinn M
Reporting of systematic reviews: the challenge of genetic association studies.
PLoS Med. 2007 Jun;4(6):e211. [Abstract/Link to Full Text]

Reis BY, Kohane IS, Mandl KD
An epidemiological network model for disease outbreak detection.
PLoS Med. 2007 Jun;4(6):e210.
BACKGROUND: Advanced disease-surveillance systems have been deployed worldwide to provide early detection of infectious disease outbreaks and bioterrorist attacks. New methods that improve the overall detection capabilities of these systems can have a broad practical impact. Furthermore, most current generation surveillance systems are vulnerable to dramatic and unpredictable shifts in the health-care data that they monitor. These shifts can occur during major public events, such as the Olympics, as a result of population surges and public closures. Shifts can also occur during epidemics and pandemics as a result of quarantines, the worried-well flooding emergency departments or, conversely, the public staying away from hospitals for fear of nosocomial infection. Most surveillance systems are not robust to such shifts in health-care utilization, either because they do not adjust baselines and alert-thresholds to new utilization levels, or because the utilization shifts themselves may trigger an alarm. As a result, public-health crises and major public events threaten to undermine health-surveillance systems at the very times they are needed most. METHODS AND FINDINGS: To address this challenge, we introduce a class of epidemiological network models that monitor the relationships among different health-care data streams instead of monitoring the data streams themselves. By extracting the extra information present in the relationships between the data streams, these models have the potential to improve the detection capabilities of a system. Furthermore, the models' relational nature has the potential to increase a system's robustness to unpredictable baseline shifts. We implemented these models and evaluated their effectiveness using historical emergency department data from five hospitals in a single metropolitan area, recorded over a period of 4.5 y by the Automated Epidemiological Geotemporal Integrated Surveillance real-time public health-surveillance system, developed by the Children's Hospital Informatics Program at the Harvard-MIT Division of Health Sciences and Technology on behalf of the Massachusetts Department of Public Health. We performed experiments with semi-synthetic outbreaks of different magnitudes and simulated baseline shifts of different types and magnitudes. The results show that the network models provide better detection of localized outbreaks, and greater robustness to unpredictable shifts than a reference time-series modeling approach. CONCLUSIONS: The integrated network models of epidemiological data streams and their interrelationships have the potential to improve current surveillance efforts, providing better localized outbreak detection under normal circumstances, as well as more robust performance in the face of shifts in health-care utilization during epidemics and major public events. [Abstract/Link to Full Text]

Recent Articles in BMC Medicine

Odenwald M, Neuner F, Schauer M, Elbert T, Catani C, Lingenfelder B, Hinkel H, H�fner H, Rockstroh B
Khat use as risk factor for psychotic disorders: a cross-sectional and case-control study in Somalia.
BMC Med. 2005;35.
BACKGROUND: Little is known about the prevalence of khat-induced psychotic disorders in East African countries, where the chewing of khat leaves is common. Its main psycho-active component cathinone produces effects similar to those of amphetamine. We aimed to explore the prevalence of psychotic disorders among the general population and the association between khat use and psychotic symptoms. METHODS: In an epidemiological household assessment in the city of Hargeisa, North-West Somalia, trained local interviewers screened 4,854 randomly selected persons from among the general population for disability due to severe mental problems. The identified cases were interviewed based on a structured interview and compared to healthy matched controls. Psychotic symptoms were assessed using the items of the WHO Composite International Diagnostic Interview and quantified with the Positive and Negative Symptoms Scale. Statistical testing included Student's t-test and ANOVA. RESULTS: Local interviewers found that rates of severe disability due to mental disorders were 8.4% among males (above the age of 12) and differed according to war experiences (no war experience: 3.2%; civilian war survivors: 8.0%; ex-combatants: 15.9%). The clinical interview verified that in 83% of positive screening cases psychotic symptoms were the most prominent manifestations of psychiatric illness. On average, cases with psychotic symptoms had started to use khat earlier in life than matched controls and had been using khat 8.6 years before positive symptoms emerged. In most cases with psychotic symptoms, a pattern of binge use (> two 'bundles' per day) preceded the onset of psychotic symptoms, in contrast to controls of the same age. We found significant correlations between variables of khat consumption and clinical scales (0.35 to 0.50; p < 0.05), and between the age of onset of khat chewing and symptom onset (0.70; p <0.001). CONCLUSION: Evidence indicates a relationship between the consumption of khat and the onset of psychotic symptoms among the male population, whereby not the khat intake per se but rather early onset and excessive khat chewing seemed to be related to psychotic symptoms. The khat problem must be addressed by means other than prohibition, given the widespread use and its role in Somali culture. [Abstract/Link to Full Text]

Anderson ME, Johnson DC, Batal HA
Sudden Infant Death Syndrome and prenatal maternal smoking: rising attributed risk in the Back to Sleep era.
BMC Med. 2005;34.
BACKGROUND: Parental smoking and prone sleep positioning are recognized causal features of Sudden Infant Death. This study quantifies the relationship between prenatal smoking and infant death over the time period of the Back to Sleep campaign in the United States, which encouraged parents to use a supine sleeping position for infants. METHODS: This retrospective cohort study utilized the Colorado Birth Registry. All singleton, normal birth weight infants born from 1989 to 1998 were identified and linked to the Colorado Infant Death registry. Multivariable logistic regression was used to analyze the relationship between outcomes of interest and prenatal maternal cigarette use. Potential confounders analyzed included infant gender, gestational age, and birth year as well as maternal marital status, ethnicity, pregnancy interval, age, education, and alcohol use. RESULTS: We analyzed 488,918 birth records after excluding 5835 records with missing smoking status. Smokers were more likely to be single, non-Hispanic, less educated, and to report alcohol use while pregnant (p < 0.001). The study included 598 SIDS cases of which 172 occurred in smoke-exposed infants. Smoke exposed infants were 1.9 times (95% CI 1.6 to 2.3) more likely to die of SIDS. The attributed risk associating smoking and SIDS increased during the study period from approximately 50% to 80%. During the entire study period 59% (101/172) of SIDS deaths in smoke-exposed infants were attributed to maternal smoking. CONCLUSIONS: Due to a decreased overall rate of SIDS likely due to changing infant sleep position, the attributed risk associating maternal smoking and SIDS has increased following the Back to Sleep campaign. Mothers should be informed of the 2-fold increased rate of SIDS associated with maternal cigarette consumption. [Abstract/Link to Full Text]

Tomson J, Lip GY
Blood pressure demographics: nature or nurture ... ... genes or environment?
BMC Med. 2005;33.
Hypertension is a growing worldwide problem associated with an increased risk of cardiovascular morbidity and mortality. However, the rates of prevalence of hypertension are higher in some populations than others. Although ethnic and genetic factors have been implied in the past to explain this, the environmental influence and psychosocial factors may play a more important role than is widely accepted. Examining the non-genetic influences in future hypertension research may be necessary in order to clearly define the local blood pressure demographics and the global hypertensive disease burden. [Abstract/Link to Full Text]

Cooper RS, Wolf-Maier K, Luke A, Adeyemo A, Banegas JR, Forrester T, Giampaoli S, Joffres M, Kastarinen M, Primatesta P, Stegmayr B, Thamm M
An international comparative study of blood pressure in populations of European vs. African descent.
BMC Med. 2005;32.
BACKGROUND: The consistent finding of higher prevalence of hypertension in US blacks compared to whites has led to speculation that African-origin populations are particularly susceptible to this condition. Large surveys now provide new information on this issue. METHODS: Using a standardized analysis strategy we examined prevalence estimates for 8 white and 3 black populations (N = 85,000 participants). RESULTS: The range in hypertension prevalence was from 27 to 55% for whites and 14 to 44% for blacks. CONCLUSIONS: These data demonstrate that not only is there a wide variation in hypertension prevalence among both racial groups, the rates among blacks are not unusually high when viewed internationally. These data suggest that the impact of environmental factors among both populations may have been under-appreciated. [Abstract/Link to Full Text]

Champagne J, Geelen P, Philippon F, Brugada P
Recurrent cardiac events in patients with idiopathic ventricular fibrillation, excluding patients with the Brugada syndrome.
BMC Med. 2005;31.
BACKGROUND: The recurrence of cardiac events in patients with idiopathic ventricular fibrillation (VF) excluding patients with the Brugada syndrome is unclear since this entity remains present in previous studies. METHODS: Since 1992, 18 patients (72% male) with idiopathic VF out of 455 ICD implants were treated with an implantable cardioverter defibrillator (ICD). The mean age at first ICD implantation was 42 +/- 14 years. Brugada syndrome, as well as other primary electrical diseases (e.g. long QT), were systematically excluded in all patients by the absence of the typical electrocardiogram (ST elevation in the right precordial leads) at rest and/or after pharmacological tests (ajmaline, flecainide, or procainamide). Recurrence of cardiac events was prospectively assessed. RESULTS: During a mean follow-up period of 41 +/- 27 months, VF recurrence with appropriate shock occurred in 7 patients (39%) covering a total of 27 shocks. The median time to first appropriate shock was 12 +/- 9 months. There were no deaths. In the electrophysiological study, 39% of patients were inducible, but inducibility failed to predict subsequent arrhythmic events. Forty-four percent of patients suffered 21 inappropriate shocks, which were caused by sinus tachycardia, atrial arrhythmias or lead malfunction. CONCLUSION: Idiopathic ventricular fibrillation patients have a high recurrence rate of potentially fatal ventricular arrhythmias, excluding patients with the Brugada syndrome or other known causes. ICD prevents sudden cardiac death but inappropriate shocks remained a major issue in this young and active population. [Abstract/Link to Full Text]

Smeeth L, Cook C, Fombonne PE, Heavey L, Rodrigues LC, Smith PG, Hall AJ
Rate of first recorded diagnosis of autism and other pervasive developmental disorders in United Kingdom general practice, 1988 to 2001.
BMC Med. 2004 Nov 9;239.
BACKGROUND: There has been concern that the incidence of autism and other pervasive developmental disorders (PDDs) is increasing. Previous studies have been smaller, restricted to autism (excluding other pervasive developmental disorders such as Asperger's syndrome), included boys only, or have not been based on a national sample. We investigated time trends in the rates of diagnosis of pervasive developmental disorders. METHODS: We analysed the rates of first diagnosis of pervasive developmental disorders among people registered with a practice contributing to the United Kingdom General Practice Research Database during the period 1988 to 2001. We included 1410 cases from over 14 million person-years of observation. The main outcome measures were rates of diagnosis of pervasive developmental disorders by year of diagnosis, year of birth, gender and geographical region. RESULTS: The rate increased progressively from 0.40/10,000 person-years (95% CI 0.30 to 0.54) in 1991 to 2.98/10,000 (95% CI 2.56 to 3.47) in 2001. A similar change occurred in the age standardised incidence ratios, from 35 (95% CI: 26-47) in 1991 to 365 (95% CI: 314-425) in 2001. The temporal increase was not limited to children born during specific years nor to children diagnosed in a specific time period. The rate of diagnosis of PDDs other than autism rose from zero for the period 1988-1992 to 1.06/10,000 person-years in 2001. The rate of diagnosis of autism also increased but to a lesser extent. There was marked geographical variation in rates, with standardised incidence ratios varying from 66 for Wales to 141 for the South East of England. CONCLUSIONS: Better ascertainment of diagnosis is likely to have contributed to the observed temporal increase in rates of diagnosis of PDD, but we cannot exclude a real increase. [Abstract/Link to Full Text]

Bagshaw SM, Ghali WA
Acetylcysteine for prevention of contrast-induced nephropathy after intravascular angiography: a systematic review and meta-analysis.
BMC Med. 2004 Oct 22;238.
BACKGROUND: Contrast-induced nephropathy is an important cause of acute renal failure. We assess the efficacy of acetylcysteine for prevention of contrast-induced nephropathy among patients undergoing intravascular angiography. METHODS: We conducted a systematic review and meta-analysis of randomized controlled trials comparing prophylactic acetylcysteine plus hydration versus hydration alone in patients undergoing intravascular angiography. Studies were identified by searching MEDLINE, EMBASE, and CENTRAL databases. Our main outcome measures were the risk of contrast-induced nephropathy and the difference in serum creatinine between acetylcysteine and control groups at 48 h. RESULTS: Fourteen studies involving 1261 patients were identified and included for analysis, and findings were heterogeneous across studies. Acetylcysteine was associated with a significantly reduced incidence of contrast-induced nephropathy in five studies, and no difference in the other nine (with a trend toward a higher incidence in six of the latter studies). The pooled odds ratio for contrast-induced nephropathy with acetylcysteine relative to control was 0.54 (95% CI, 0.32-0.91, p = 0.02) and the pooled estimate of difference in 48-h serum creatinine for acetylcysteine relative to control was -7.2 mumol/L (95% CI -19.7 to 5.3, p = 0.26). These pooled values need to be interpreted cautiously because of the heterogeneity across studies, and due to evidence of publication bias. Meta-regression suggested that the heterogeneity might be partially explained by whether the angiography was performed electively or as emergency. CONCLUSION: These findings indicate that published studies of acetylcysteine for prevention of contrast-induced nephropathy yield inconsistent results. The efficacy of acetylcysteine will remain uncertain unless a large well-designed multi-center trial is performed. [Abstract/Link to Full Text]

Marchi N, Hallene KL, Kight KM, Cucullo L, Moddel G, Bingaman W, Dini G, Vezzani A, Janigro D
Significance of MDR1 and multiple drug resistance in refractory human epileptic brain.
BMC Med. 2004 Oct 9;237.
BACKGROUND: The multiple drug resistance protein (MDR1/P-glycoprotein) is overexpressed in glia and blood-brain barrier (BBB) endothelium in drug refractory human epileptic tissue. Since various antiepileptic drugs (AEDs) can act as substrates for MDR1, the enhanced expression/function of this protein may increase their active extrusion from the brain, resulting in decreased responsiveness to AEDs. METHODS: Human drug resistant epileptic brain tissues were collected after surgical resection. Astrocyte cell cultures were established from these tissues, and commercially available normal human astrocytes were used as controls. Uptake of fluorescent doxorubicin and radioactive-labeled Phenytoin was measured in the two cell populations, and the effect of MDR1 blockers was evaluated.Frozen human epileptic brain tissue slices were double immunostained to locate MDR1 in neurons and glia. Other slices were exposed to toxic concentrations of Phenytoin to study cell viability in the presence or absence of a specific MDR1 blocker. RESULTS: MDR1 was overexpressed in blood vessels, astrocytes and neurons in human epileptic drug-resistant brain. In addition, MDR1-mediated cellular drug extrusion was increased in human 'epileptic' astrocytes compared to 'normal' ones. Concomitantly, cell viability in the presence of cytotoxic compounds was increased. CONCLUSIONS: Overexpression of MDR1 in different cell types in drug-resistant epileptic human brain leads to functional alterations, not all of which are linked to drug pharmacokinetics. In particular, the modulation of glioneuronal MDR1 function in epileptic brain in the presence of toxic concentrations of xenobiotics may constitute a novel cytoprotective mechanism. [Abstract/Link to Full Text]

Booth M, Vennervald BJ, Butterworth AE, Kariuki HC, Amaganga C, Kimani G, Mwatha JK, Otedo A, Ouma JH, Dunne DW
Exposure to malaria affects the regression of hepatosplenomegaly after treatment for Schistosoma mansoni infection in Kenyan children.
BMC Med. 2004 Sep 27;236.
BACKGROUND: Schistosoma mansoni and malaria infections are often endemic in the same communities in sub-Saharan Africa, and both have pathological effects on the liver and the spleen. Hepatosplenomegaly associated with S. mansoni is exacerbated in children with relatively high exposure to malaria. Treatment with praziquantel reduces the degree of hepatosplenomegaly, but the condition does not completely resolve in some cases. The present analysis focused on the possibility that exposure to malaria infection may have limited the resolution of hepatosplenomegaly in a cohort of Kenyan schoolchildren. METHODS: Ninety-six children aged 6-16, from one community in Makueni district, Kenya, were treated with praziquantel. At baseline, all children had hepatomegaly and most had splenomegaly. The source of S. mansoni infection, a river, was molluscicided regularly over the following three years to limit S. mansoni re-infection, whereas malaria exposure was uninterrupted. Hepatic and splenic enlargement was assessed annually outside the malaria transmission season. RESULTS: Children living in an area of relatively high exposure to both infections presented with the largest spleens before treatment and at each follow-up. Spleens of firm consistency were associated with proximity to the river. The regression of hepatomegaly was also affected by location, being minimal in an area with relatively low S. mansoni exposure but high exposure to malaria, and maximal in an area with relatively low exposure to both infections. CONCLUSIONS: The outcome of treating cases of hepatosplenomegaly with praziquantel in this cohort of Kenyan children depended strongly on their level of exposure to malaria infection. Furthermore, a residual burden of hepatosplenic morbidity was observed, which was possibly attributable to the level of exposure to malaria. The results suggest that exposure to malaria infection may be a significant factor affecting the outcome of praziquantel treatment to reduce the level of hepatosplenic morbidity. [Abstract/Link to Full Text]

Malthaner RA, Wong RK, Rumble RB, Zuraw L
Neoadjuvant or adjuvant therapy for resectable esophageal cancer: a systematic review and meta-analysis.
BMC Med. 2004 9 24;2(1):35.
BACKGROUND: Carcinoma of the esophagus is an aggressive malignancy with an increasing incidence. Its virulence, in terms of symptoms and mortality, justifies a continued search for optimal therapy. The large and growing number of patients affected, the high mortality rates, the worldwide geographic variation in practice, and the large body of good quality research warrants a systematic review with meta-analysis. METHODS: A systematic review and meta-analysis investigating the impact of neoadjuvant or adjuvant therapy on resectable thoracic esophageal cancer to inform evidence-based practice was produced.MEDLINE, CANCERLIT, Cochrane Library, EMBASE, and abstracts from the American Society of Clinical Oncology and the American Society for Therapeutic Radiology and Oncology were searched for trial reports.Included were randomized trials or meta-analyses of neoadjuvant or adjuvant treatments compared with surgery alone or other treatments in patients with resectable thoracic esophageal cancer. Outcomes of interest were survival, adverse effects, and quality of life. Either one- or three-year mortality data were pooled and reported as relative risk ratios. RESULTS: Thirty-four randomized controlled trials and six meta-analyses were obtained and grouped into 13 basic treatment approaches.Single randomized controlled trials detected no differences in mortality between treatments for the following comparisons:- Preoperative radiotherapy versus postoperative radiotherapy.- Preoperative and postoperative radiotherapy versus postoperative radiotherapy. Preoperative and postoperative radiotherapy was associated with a significantly higher mortality rate.- Postoperative chemotherapy versus postoperative radiotherapy.- Postoperative radiotherapy versus postoperative radiotherapy plus protein-bound polysaccharide versus chemoradiation versus chemoradiation plus protein-bound polysaccharide.Pooling one-year mortality detected no statistically significant differences in mortality between treatments for the following comparisons:- Preoperative radiotherapy compared with surgery alone (five randomized trials).- Postoperative radiotherapy compared with surgery alone (five randomized trials).- Preoperative chemotherapy versus surgery alone (six randomized trials).- Preoperative and postoperative chemotherapy versus surgery alone (two randomized trials).- Preoperative chemoradiation therapy versus surgery alone (six randomized trials).Single randomized controlled trials detected differences in mortality between treatments for the following comparison:- Preoperative hyperthermia and chemoradiotherapy versus preoperative chemoradiotherapy in favour of hyperthermia.Pooling three-year mortality detected no statistically significant difference in mortality between treatments for the following comparison:- Postoperative chemotherapy compared with surgery alone (two randomized trials).Pooling three-year mortality detected statistically significant differences between treatments for the following comparisons:- Preoperative chemoradiation therapy versus surgery alone (six randomized trials) in favour of preoperative chemoradiation with surgery.- Preoperative chemotherapy compared with preoperative radiotherapy (one randomized trial) in favour of preoperative radiotherapy. CONCLUSION: For adult patients with resectable thoracic esophageal cancer for whom surgery is considered appropriate, surgery alone (i.e., without neoadjuvant or adjuvant therapy) is recommended as the standard practice. [Abstract/Link to Full Text]

Medveczky MM, Sherwood TA, Klein TW, Friedman H, Medveczky PG
Delta-9 tetrahydrocannabinol (THC) inhibits lytic replication of gamma oncogenic herpesviruses in vitro.
BMC Med. 2004 Sep 15;234.
BACKGROUND: The major psychoactive cannabinoid compound of marijuana, delta-9 tetrahydrocannabinol (THC), has been shown to modulate immune responses and lymphocyte function. After primary infection the viral DNA genome of gamma herpesviruses persists in lymphoid cell nuclei in a latent episomal circular form. In response to extracellular signals, the latent virus can be activated, which leads to production of infectious virus progeny. Therefore, we evaluated the potential effects of THC on gamma herpesvirus replication. METHODS: Tissue cultures infected with various gamma herpesviruses were cultured in the presence of increasing concentrations of THC and the amount of viral DNA or infectious virus yield was compared to those of control cultures. The effect of THC on Kaposi's Sarcoma Associated Herpesvirus (KSHV) and Epstein-Barr virus (EBV) replication was measured by the Gardella method and replication of herpesvirus saimiri (HVS) of monkeys, murine gamma herpesvirus 68 (MHV 68), and herpes simplex type 1 (HSV-1) was measured by yield reduction assays. Inhibition of the immediate early ORF 50 gene promoter activity was measured by the dual luciferase method. RESULTS: Micromolar concentrations of THC inhibit KSHV and EBV reactivation in virus infected/immortalized B cells. THC also strongly inhibits lytic replication of MHV 68 and HVS in vitro. Importantly, concentrations of THC that inhibit virus replication of gamma herpesviruses have no effect on cell growth or HSV-1 replication, indicating selectivity. THC was shown to selectively inhibit the immediate early ORF 50 gene promoter of KSHV and MHV 68. CONCLUSIONS: THC specifically targets viral and/or cellular mechanisms required for replication and possibly shared by these gamma herpesviruses, and the endocannabinoid system is possibly involved in regulating gamma herpesvirus latency and lytic replication. The immediate early gene ORF 50 promoter activity was specifically inhibited by THC. These studies may also provide the foundation for the development of antiviral strategies utilizing non-psychoactive derivatives of THC. [Abstract/Link to Full Text]

McKibbon KA, Wilczynski NL, Haynes RB
What do evidence-based secondary journals tell us about the publication of clinically important articles in primary healthcare journals?
BMC Med. 2004 Sep 6;233.
BACKGROUND: We conducted this analysis to determine i) which journals publish high-quality, clinically relevant studies in internal medicine, general/family practice, general practice nursing, and mental health; and ii) the proportion of clinically relevant articles in each journal. METHODS: We performed an analytic survey of a hand search of 170 general medicine, general healthcare, and specialty journals for 2000. Research staff assessed individual articles by using explicit criteria for scientific merit for healthcare application. Practitioners assessed the clinical importance of these articles. Outcome measures were the number of high-quality, clinically relevant studies published in the 170 journal titles and how many of these were published in each of four discipline-specific, secondary "evidence-based" journals (ACP Journal Club for internal medicine and its subspecialties; Evidence-Based Medicine for general/family practice; Evidence-Based Nursing for general practice nursing; and Evidence-Based Mental Health for all aspects of mental health). Original studies and review articles were classified for purpose: therapy and prevention, screening and diagnosis, prognosis, etiology and harm, economics and cost, clinical prediction guides, and qualitative studies. RESULTS: We evaluated 60,352 articles from 170 journal titles. The pass criteria of high-quality methods and clinically relevant material were met by 3059 original articles and 1073 review articles. For ACP Journal Club (internal medicine), four titles supplied 56.5% of the articles and 27 titles supplied the other 43.5%. For Evidence-Based Medicine (general/family practice), five titles supplied 50.7% of the articles and 40 titles supplied the remaining 49.3%. For Evidence-Based Nursing (general practice nursing), seven titles supplied 51.0% of the articles and 34 additional titles supplied 49.0%. For Evidence-Based Mental Health (mental health), nine titles supplied 53.2% of the articles and 34 additional titles supplied 46.8%. For the disciplines of internal medicine, general/family practice, and mental health (but not general practice nursing), the number of clinically important articles was correlated withScience Citation Index (SCI) Impact Factors. CONCLUSIONS: Although many clinical journals publish high-quality, clinically relevant and important original studies and systematic reviews, the articles for each discipline studied were concentrated in a small subset of journals. This subset varied according to healthcare discipline; however, many of the important articles for all disciplines in this study were published in broad-based healthcare journals rather than subspecialty or discipline-specific journals. [Abstract/Link to Full Text]

Nyg�rd K, de Jong B, Guerin PJ, Andersson Y, Olsson A, Giesecke J
Emergence of new Salmonella Enteritidis phage types in Europe? Surveillance of infections in returning travellers.
BMC Med. 2004 Sep 2;232.
BACKGROUND: Among human Salmonella Enteritidis infections, phage type 4 has been the dominant phage type in most countries in Western Europe during the last years. This is reflected in Salmonella infections among Swedish travellers returning from abroad. However, there are differences in phage type distribution between the countries, and this has also changed over time. METHODS: We used data from the Swedish infectious disease register and the national reference laboratory to describe phage type distribution of Salmonella Enteritidis infections in Swedish travellers from 1997 to 2002, and have compared this with national studies conducted in the countries visited. RESULTS: Infections among Swedish travellers correlate well with national studies conducted in the countries visited. In 2001 a change in phage type distribution in S. Enteritidis infections among Swedish travellers returning from some countries in southern Europe was observed, and a previously rare phage type (PT 14b) became one of the most commonly diagnosed that year, continuing into 2002 and 2003. CONCLUSIONS: Surveillance of infections among returning travellers can be helpful in detecting emerging infections and outbreaks in tourist destinations. The information needs to be communicated rapidly to all affected countries in order to expedite the implementation of appropriate investigations and preventive measures. [Abstract/Link to Full Text]

Pletcher MJ, Tice JA, Pignone M, McCulloch C, Callister TQ, Browner WS
What does my patient's coronary artery calcium score mean? Combining information from the coronary artery calcium score with information from conventional risk factors to estimate coronary heart disease risk.
BMC Med. 2004 Aug 24;231.
BACKGROUND: The coronary artery calcium (CAC) score is an independent predictor of coronary heart disease. We sought to combine information from the CAC score with information from conventional cardiac risk factors to produce post-test risk estimates, and to determine whether the score may add clinically useful information. METHODS: We measured the independent cross-sectional associations between conventional cardiac risk factors and the CAC score among asymptomatic persons referred for non-contrast electron beam computed tomography. Using the resulting multivariable models and published CAC score-specific relative risk estimates, we estimated post-test coronary heart disease risk in a number of different scenarios. RESULTS: Among 9341 asymptomatic study participants (age 35-88 years, 40% female), we found that conventional coronary heart disease risk factors including age, male sex, self-reported hypertension, diabetes and high cholesterol were independent predictors of the CAC score, and we used the resulting multivariable models for predicting post-test risk in a variety of scenarios. Our models predicted, for example, that a 60-year-old non-smoking non-diabetic women with hypertension and high cholesterol would have a 47% chance of having a CAC score of zero, reducing her 10-year risk estimate from 15% (per Framingham) to 6-9%; if her score were over 100, however (a 17% chance), her risk estimate would be markedly higher (25-51% in 10 years). In low risk scenarios, the CAC score is very likely to be zero or low, and unlikely to change management. CONCLUSION: Combining information from the CAC score with information from conventional risk factors can change assessment of coronary heart disease risk to an extent that may be clinically important, especially when the pre-test 10-year risk estimate is intermediate. The attached spreadsheet makes these calculations easy. [Abstract/Link to Full Text]

Wangemann P, Itza EM, Albrecht B, Wu T, Jabba SV, Maganti RJ, Lee JH, Everett LA, Wall SM, Royaux IE, Green ED, Marcus DC
Loss of KCNJ10 protein expression abolishes endocochlear potential and causes deafness in Pendred syndrome mouse model.
BMC Med. 2004 Aug 20;230.
BACKGROUND: Pendred syndrome, a common autosomal-recessive disorder characterized by congenital deafness and goiter, is caused by mutations of SLC26A4, which codes for pendrin. We investigated the relationship between pendrin and deafness using mice that have (Slc26a4+/+) or lack a complete Slc26a4 gene (Slc26a4-/-). METHODS: Expression of pendrin and other proteins was determined by confocal immunocytochemistry. Expression of mRNA was determined by quantitative RT-PCR. The endocochlear potential and the endolymphatic K+ concentration were measured with double-barreled microelectrodes. Currents generated by the stria marginal cells were recorded with a vibrating probe. Tissue masses were evaluated by morphometric distance measurements and pigmentation was quantified by densitometry. RESULTS: Pendrin was found in the cochlea in apical membranes of spiral prominence cells and spindle-shaped cells of stria vascularis, in outer sulcus and root cells. Endolymph volume in Slc26a4-/- mice was increased and tissue masses in areas normally occupied by type I and II fibrocytes were reduced. Slc26a4-/- mice lacked the endocochlear potential, which is generated across the basal cell barrier by the K+ channel KCNJ10 localized in intermediate cells. Stria vascularis was hyperpigmented, suggesting unalleviated free radical damage. The basal cell barrier appeared intact; intermediate cells and KCNJ10 mRNA were present but KCNJ10 protein was absent. Endolymphatic K+ concentrations were normal and membrane proteins necessary for K+ secretion were present, including the K+ channel KCNQ1 and KCNE1, Na+/2Cl-/K+ cotransporter SLC12A2 and the gap junction GJB2. CONCLUSIONS: These observations demonstrate that pendrin dysfunction leads to a loss of KCNJ10 protein expression and a loss of the endocochlear potential, which may be the direct cause of deafness in Pendred syndrome. [Abstract/Link to Full Text]

McManus IC, Keeling A, Paice E
Stress, burnout and doctors' attitudes to work are determined by personality and learning style: a twelve year longitudinal study of UK medical graduates.
BMC Med. 2004 Aug 18;229.
BACKGROUND: The study investigated the extent to which approaches to work, workplace climate, stress, burnout and satisfaction with medicine as a career in doctors aged about thirty are predicted by measures of learning style and personality measured five to twelve years earlier when the doctors were applicants to medical school or were medical students. METHODS: Prospective study of a large cohort of doctors. The participants were first studied when they applied to any of five UK medical schools in 1990. Postal questionnaires were sent to all doctors with a traceable address on the current or a previous Medical Register. The current questionnaire included measures of Approaches to Work, Workplace Climate, stress (General Health Questionnaire), burnout (Maslach Burnout Inventory), and satisfaction with medicine as a career and personality (Big Five). Previous questionnaires had included measures of learning style (Study Process Questionnaire) and personality. RESULTS: Doctors' approaches to work were predicted by study habits and learning styles, both at application to medical school and in the final year. How doctors perceive their workplace climate and workload is predicted both by approaches to work and by measures of stress, burnout and satisfaction with medicine. These characteristics are partially predicted by trait measures of personality taken five years earlier. Stress, burnout and satisfaction also correlate with trait measures of personality taken five years earlier. CONCLUSIONS: Differences in approach to work and perceived workplace climate seem mainly to reflect stable, long-term individual differences in doctors themselves, reflected in measures of personality and learning style. [Abstract/Link to Full Text]

Sastry J, Pisal H, Sutar S, Kapadia-Kundu N, Joshi A, Suryavanshi N, Bharucha KE, Shrotri A, Phadke MA, Bollinger RC, Shankar AV
Optimizing the HIV/AIDS informed consent process in India.
BMC Med. 2004 Aug 2;228.
BACKGROUND: While the basic ethical issues regarding consent may be universal to all countries, the consent procedures required by international review boards which include detailed scientific and legal information, may not be optimal when administered within certain populations. The time and the technicalities of the process itself intimidate individuals in societies where literacy and awareness about medical and legal rights is low. METHODS: In this study, we examined pregnant women's understanding of group education and counseling (GEC) about HIV/AIDS provided within an antenatal clinic in Maharashtra, India. We then enhanced the GEC process with the use of culturally appropriate visual aids and assessed the subsequent changes in women's understanding of informed consent issues. RESULTS: We found the use of visual aids during group counseling sessions increased women's overall understanding of key issues regarding informed consent from 38% to 72%. Moreover, if these same visuals were reinforced during individual counseling, improvements in women's overall comprehension rose to 96%. CONCLUSIONS: This study demonstrates that complex constructs such as informed consent can be conveyed in populations with little education and within busy government hospital settings, and that the standard model may not be sufficient to ensure true informed consent. [Abstract/Link to Full Text]

Smeeton NC, Rona RJ, Dobson P, Cochrane R, Wolfe C
Assessing the determinants of stillbirths and early neonatal deaths using routinely collected data in an inner city area.
BMC Med. 2004 Jul 6;227.
BACKGROUND: Within the UK there is considerable variation in the perinatal mortality rate. The objective of this study was to assess the factors associated with stillbirths and early neonatal deaths (ENND) and the suitability of the available databases in a health authority with one of the highest rates in the country. METHODS: Two case-control studies were carried out in three hospital trusts in the Lambeth, Southwark and Lewisham Health Authority, London, using routinely collected information. In one study, 342 stillbirths and 1,368 controls were included, and in the other study, 205 ENND and 820 controls were included. In the two studies cases and controls were matched for hospital trust. RESULTS: A birthweight below 1.5 kg was found in 54% and 48% of the stillbirths and ENND, respectively. More than 50% of the cases, stillbirths and ENND, had a length of gestation below 32 weeks. Length of gestation, birthweight, emergency caesarean section and age of the mother were associated with stillbirths. Birthweight and Apgar score at 1 minute as a categorical variable were associated with ENND. There was no direct evidence of an effect of social deprivation on the outcomes of interest. CONCLUSION: Birthweight and length of gestation are the most influential factors on an unfavourable outcome. Conception at an older age has a serious impact on stillbirth rates. In our health authority social disadvantage did not have a direct impact on stillbirth and ENND. Maternity information systems should collect routine data on fewer variables, but their quality in terms of value, standardization and completion rates must improve. [Abstract/Link to Full Text]

Lauer JA, Betr�n AP, Victora CG, de On�s M, Barros AJ
Breastfeeding patterns and exposure to suboptimal breastfeeding among children in developing countries: review and analysis of nationally representative surveys.
BMC Med. 2004 Jul 1;226.
BACKGROUND: Suboptimal breastfeeding is associated with higher mortality among infants and young children in the developing world. We describe patterns in 'exclusive breastfeeding' and 'any breastfeeding' rates and quantify exposure to suboptimal breastfeeding among children aged two years or younger in developing countries. METHODS: We reviewed nationally representative surveys that collected data on breastfeeding rates in 94 developing countries. Surveys were categorized by completeness and comprehensiveness of data. Complete and comprehensive data were analysed with minimum chi-square regression. With a fitting procedure, estimated parameters were used to impute missing observations for incomplete or non-comprehensive surveys. Breastfeeding indicators were calculated and are reported for 135 developing countries by UN region. RESULTS: Amongst infants aged six months or younger in the developing world, the prevalence of exclusive breastfeeding is 39% and the prevalence of no breastfeeding is 5.6%. The prevalence of continued breastfeeding is 86% and 68% for infants and children aged 6-11 and 12-23 months, respectively, in the developing world. Imputation expands population coverage of indicators, especially for infants. Breastfeeding trends are highly linear and estimated parameters defining the age-specific attrition hazard are robust. Survey-reported rates, particularly for exclusive breastfeeding, appear to have systematic upward bias, and exposure estimates must be considered conservative. CONCLUSIONS: Compliance with breastfeeding recommendations in developing countries is low, and more attention should be given to increasing breastfeeding - especially exclusive breastfeeding - and to monitoring trends. Although the introduction of more standardized and better validated survey instruments is desirable, since data coverage, completeness and comprehensiveness are extensive, global exposure assessment is relatively robust. Moreover, the regularity of breastfeeding patterns show existing survey data capture real biological and social phenomena. Our method for the analysis of breastfeeding rates provides a potent tool for summarizing trends, validating observations, translating and extrapolating indicators (as well as projecting and imputing estimates when necessary) and should support more effective child health monitoring. [Abstract/Link to Full Text]

Wahl RA, Sisk DJ, Ball TM
Clinic-based screening for domestic violence: use of a child safety questionnaire.
BMC Med. 2004 Jun 30;225.
BACKGROUND: Domestic violence affects many women during their lifetime. Children living in homes where they are or have been exposed to violence are at increased risk for adverse outcomes. The American Academy of Pediatrics, the American Academy of Family Practice, and the American College of Obstetrics/Gynecology have recently joined in recommending routine screening of all families for the presence of domestic violence. We present our experience with an office-based domestic violence screening questionnaire. METHODS: A series of four child safety questionnaires (designed for parents of infant, preschool-age, school-age, and adolescent patients), which included specific questions about domestic violence, was given to all mothers presenting to a university out-patient general pediatric clinic. The questionnaires, offered in both English and Spanish, were reviewed for the presence of domestic violence exposure, usually at the time of the clinic visit. The number of women who reported either current or past exposure to domestic violence as disclosed by this active screening process was compared to the number discovered prior to the use of these questionnaires. RESULTS: Prior to the use of active screening with a child safety questionnaire, five cases of domestic violence were identified in our clinic population of approximately 5000 children over a 3 month period. Active screening of this population with a parent questionnaire resulted in the identification of 69 cases of current domestic violence exposure (2% of those screened) during each of 2 years of screening. Use of the child safety questionnaire was associated with a significantly increased odds of detecting current domestic violence (OR = 3.6, 95% CI [1.4, 9.1], P = 0.007), with 72% [26-84%] of the cases identified being attributable to the use of the questionnaire. Of children screened, 2% were currently exposed to domestic violence, and 13% had been exposed to past domestic violence. Thus a total of 15% of our patient population has been exposed to domestic violence in their homes. CONCLUSION: Children in our clinic population are frequently exposed to domestic violence. Active screening for the presence of current or past domestic violence through the use of a parent questionnaire resulted in a significant increase in our ability to identify such families and provide appropriate referral information. [Abstract/Link to Full Text]

Podsypanina K, Li Y, Varmus HE
Evolution of somatic mutations in mammary tumors in transgenic mice is influenced by the inherited genotype.
BMC Med. 2004 Jun 15;224.
BACKGROUND: MMTV-Wnt1 transgenic mice develop mammary hyperplasia early in development, followed by the appearance of solitary mammary tumors with a high proportion of cells expressing early lineage markers and many myoepithelial cells. The occurrence of tumors is accelerated in experiments that activate FGF proto-oncogenes or remove the tumor suppressor genes Pten or P53, implying that secondary oncogenic events are required for progression from mammary hyperplasia to carcinoma. It is not known, however, which oncogenic pathways contribute to Wnt1-induced tumorigenesis - further experimental manipulation of these mice is needed. Secondary events also appear to be required for mammary tumorigenesis in MMTV-Neu transgenic mice because the transgene in the tumors usually contains an acquired mutation that activates the Neu protein-tyrosine kinase. METHODS: cDNA or DNA from the mammary glands and mammary tumors from MMTV-Wnt1, MMTV-Wnt1/p53-/-, MMTV-Neu transgenic mice, and newly generated MMTV-Wnt1/MMTV-Neu bitransgenic mice, was sequenced to seek activating mutations in H-Ras, K-Ras, and N-Ras genes, or in the MMTV-Neu transgene. In addition, tumors from bitransgenic animals were examined to determine the cellular phenotype. RESULTS: We found activating mutations at codons 12, 13, and 61 of H-Ras in just over half of the mammary tumors in MMTV-Wnt1 transgenic mice, and we confirmed the high frequency of activating mutations of Neu in tumors in MMTV-Neu transgenic mice. Tumors appeared earlier in bitransgenic MMTV-Wnt1/MMTV-Neu mice, but no Ras or MMTV-Neu mutations were found in these tumors, which were phenotypically similar to those arising in MMTV-Wnt1 mice. In addition, no Ras mutations were found in the mammary tumors that arise in MMTV-Wnt1 transgenic mice lacking an intact P53 gene. CONCLUSIONS: Tumorigenic properties of cells undergoing functionally significant secondary mutations in H-Ras or the MMTV-Neu transgene allow selection of those cells in MMTV-Wnt1 and MMTV-Neu transgenic mice, respectively. Alternative sources of oncogenic potential, such as a second transgenic oncogene or deficiency of a tumor suppressor gene, can obviate the selective power of those secondary mutations. These observations are consistent with the notion that somatic evolution of mouse mammary tumors is influenced by the specific nature of the inherited cancer-promoting genotype. [Abstract/Link to Full Text]

Wilczynski NL, Haynes RB
Developing optimal search strategies for detecting clinically sound prognostic studies in MEDLINE: an analytic survey.
BMC Med. 2004 Jun 9;223.
BACKGROUND: Clinical end users of MEDLINE have a difficult time retrieving articles that are both scientifically sound and directly relevant to clinical practice. Search filters have been developed to assist end users in increasing the success of their searches. Many filters have been developed for the literature on therapy and reviews but little has been done in the area of prognosis. The objective of this study is to determine how well various methodologic textwords, Medical Subject Headings, and their Boolean combinations retrieve methodologically sound literature on the prognosis of health disorders in MEDLINE. METHODS: An analytic survey was conducted, comparing hand searches of journals with retrievals from MEDLINE for candidate search terms and combinations. Six research assistants read all issues of 161 journals for the publishing year 2000. All articles were rated using purpose and quality indicators and categorized into clinically relevant original studies, review articles, general papers, or case reports. The original and review articles were then categorized as 'pass' or 'fail' for methodologic rigor in the areas of prognosis and other clinical topics. Candidate search strategies were developed for prognosis and run in MEDLINE - the retrievals being compared with the hand search data. The sensitivity, specificity, precision, and accuracy of the search strategies were calculated. RESULTS: 12% of studies classified as prognosis met basic criteria for scientific merit for testing clinical applications. Combinations of terms reached peak sensitivities of 90%. Compared with the best single term, multiple terms increased sensitivity for sound studies by 25.2% (absolute increase), and increased specificity, but by a much smaller amount (1.1%) when sensitivity was maximized. Combining terms to optimize both sensitivity and specificity achieved sensitivities and specificities of approximately 83% for each. CONCLUSION: Empirically derived search strategies combining indexing terms and textwords can achieve high sensitivity and specificity for retrieving sound prognostic studies from MEDLINE. [Abstract/Link to Full Text]

Nelson BD, Fernandez WG, Galea S, Sisco S, Dierberg K, Gorgieva GS, Nandi AK, Ahern J, Mitrovi? M, VanRooyen M, Vlahov D
War-related psychological sequelae among emergency department patients in the former Republic of Yugoslavia.
BMC Med. 2004 Jun 1;222.
BACKGROUND: Residents of the Republic of Serbia faced civil war and a NATO-led bombing campaign in 1999. We sought to assess the burden of metal health dysfunction among emergency department (ED) patients presenting for care three years post-war in Serbia. METHODS: This study was conducted during July and August 2002 at two sites: a university hospital ED in Belgrade, Serbia and an ED in a remote district hospital serving a Serbian enclave in Laplje Selo, Kosovo. Investigators collected data on a systematic sample of non-acute patients presenting to the ED. All respondents completed a structured questionnaire assessing demographics and symptoms of post-traumatic stress disorder (PTSD) (using the Harvard Trauma Questionnaire), and major depression (using the Center for Epidemiologic Studies Depression Scale). RESULTS: A total of 562 respondents participated (310 in Belgrade, 252 in Laplje Selo); the response rate was 83.8%, 43% were female, and mean age was 37.6 years (SD = 13.4). Overall, 73 (13.0%) participants had symptoms consistent with PTSD, and 272 (49.2%) had symptoms consistent with depression. Sixty-six respondents had both disorders (11.9%). In separate multivariable logistic regression models, predictors of PTSD were refugee status and residence in Laplje Selo, and predictors of depression were older age, current unemployment, and lower social support. CONCLUSIONS: Three years post-war, symptoms of PTSD and major depression in Serbia remained a significant public health concern, particularly among refugees, those suffering subsequent economic instability, and persons living in rural, remote areas. [Abstract/Link to Full Text]

Malcoe LH, Duran BM, Montgomery JM
Socioeconomic disparities in intimate partner violence against Native American women: a cross-sectional study.
BMC Med. 2004 May 24;220.
BACKGROUND: Intimate partner violence (IPV) against women is a global public health problem, yet data on IPV against Native American women are extremely limited. We conducted a cross-sectional study of Native American women to determine prevalence of lifetime and past-year IPV and partner injury; examine IPV in relation to pregnancy; and assess demographic and socioeconomic correlates of past-year IPV. METHODS: Participants were recruited from a tribally-operated clinic serving low-income pregnant and childbearing women in southwest Oklahoma. A self-administered survey was completed by 312 Native American women (96% response rate) attending the clinic from June through August 1997. Lifetime and past-year IPV were measured using modified 18-item Conflict Tactics Scales. A socioeconomic index was created based on partner's education, public assistance receipt, and poverty level. RESULTS: More than half (58.7%) of participants reported lifetime physical and/or sexual IPV; 39.1% experienced severe physical IPV; 12.2% reported partner-forced sexual activity; and 40.1% reported lifetime partner-perpetrated injuries. A total of 273 women had a spouse or boyfriend during the previous 12 months (although all participants were Native American, 59.0% of partners were non-Native). Among these women, past-year prevalence was 30.1% for physical and/or sexual IPV; 15.8% for severe physical IPV; 3.3% for forced partner-perpetrated sexual activity; and 16.4% for intimate partner injury. Reported IPV prevalence during pregnancy was 9.3%. Pregnancy was not associated with past-year IPV (odds ratio = 0.9). Past-year IPV prevalence was 42.8% among women scoring low on the socioeconomic index, compared with 10.1% among the reference group. After adjusting for age, relationship status, and household size, low socioeconomic index remained strongly associated with past-year IPV (odds ratio = 5.0; 95% confidence interval: 2.4, 10.7). CONCLUSIONS: Native American women in our sample experienced exceptionally high rates of lifetime and past-year IPV. Additionally, within this low-income sample, there was strong evidence of socioeconomic variability in IPV. Further research should determine prevalence of IPV against Native American women from diverse tribes and regions, and examine pathways through which socioeconomic disadvantage may increase their IPV risk. [Abstract/Link to Full Text]

Labrecque M, Dufresne C, Barone MA, St-Hilaire K
Vasectomy surgical techniques: a systematic review.
BMC Med. 2004 May 24;221.
BACKGROUND: A wide variety of surgical techniques are used to perform vasectomy. The purpose of this systematic review was to assess if any surgical techniques to isolate or occlude the vas are associated with better outcomes in terms of occlusive and contraceptive effectiveness, and complications. METHODS: We searched MEDLINE (1966-June 2003), EMBASE (1980-June 2003), reference lists of retrieved articles, urology textbooks, and our own files looking for studies comparing two or more vasectomy surgical techniques and reporting on effectiveness and complications. From 2,058 titles or abstracts, two independent reviewers identified 224 as potentially relevant. Full reports of 219 articles were retrieved and final selection was made by the same two independent reviewers using the same criteria as for the initial selection. Discrepancies were resolved by involving a third reviewer. Data were extracted and methodological quality of selected studies was assessed by two independent reviewers. Studies were divided in broad categories (isolation, occlusion, and combined isolation and occlusion techniques) and sub-categories of specific surgical techniques performed. Qualitative analyses and syntheses were done. RESULTS: Of 31 comparative studies (37 articles), only four were randomized clinical trials, most studies were observational and retrospective. Overall methodological quality was low. From nine studies on vas isolation, there is good evidence that the no-scalpel vasectomy approach decreases the risk of surgical complications, namely hematoma/bleeding and infection, compared with incisional techniques. Five comparative studies including one high quality randomized clinical trial provided good evidence that fascial interposition (FI) increases the occlusive effectiveness of ligation and excision. Results of 11 comparative studies suggest that FI with cautery of the vas lumen provides the highest level of occlusive effectiveness, even when leaving the testicular end open. Otherwise, firm evidence to support any occlusion technique in terms of increased effectiveness or decreased risk of complications is lacking. CONCLUSIONS: Current evidence supports no-scalpel vasectomy as the safest surgical approach to isolate the vas when performing vasectomy. Adding FI increases effectiveness beyond ligation and excision alone. Occlusive effectiveness appears to be further improved by combining FI with cautery. Methodologically sound prospective controlled studies should be conducted to evaluate specific occlusion techniques further. [Abstract/Link to Full Text]

Goulter AB, Goddard MJ, Allen JC, Clark KL
ACE2 gene expression is up-regulated in the human failing heart.
BMC Med. 2004 May 19;219.
BACKGROUND: ACE2 is a novel homologue of angiotensin converting enzyme (ACE). ACE2 is highly expressed in human heart and animal data suggest that ACE2 is an essential regulator of cardiac function in vivo. Since overactivity of the renin-angiotensin system contributes to the progression of heart failure, this investigation assessed changes in gene expression of ACE2, ACE, AT1 receptor and renin in the human failing heart. METHODS: The sensitive technique of quantitative reverse transcriptase polymerase chain reaction was used to determine the level of mRNA expression of ACE and ACE2 in human ventricular myocardium from donors with non-diseased hearts (n = 9), idiopathic dilated cardiomyopathy (IDC, n = 11) and ischemic cardiomyopathy (ICM, n = 12). Following logarithmic transformation of the data, a one-way analysis of variance was performed for each target gene followed by a Dunnett's test to compare the two disease groups IDC and ICM versus control. RESULTS: As anticipated, ACE mRNA was found to be significantly increased in the failing heart with a 3.1 and 2.4-fold up-regulation found in IDC and ICM relative to non-diseased myocardium. Expression of ACE2 mRNA was also significantly up-regulated in IDC (2.4-fold increase) and ICM (1.8-fold increase) versus non-diseased myocardium. No change in angiotensin AT1 receptor mRNA expression was found in failing myocardium and renin mRNA was not detected. CONCLUSIONS: These data suggest that ACE2 is up-regulated in human IDC and ICM and are consistent with the hypothesis that differential regulation of this enzyme may have important functional consequences in heart failure. This strengthens the hypothesis that ACE2 may be a relevant target for the treatment of heart failure and will hopefully spur further studies to clarify the functional effects in human myocardium of ACE2 derived peptides. [Abstract/Link to Full Text]

Clark TJ, ter Riet G, Coomarasamy A, Khan KS
Bias associated with delayed verification in test accuracy studies: accuracy of tests for endometrial hyperplasia may be much higher than we think!
BMC Med. 2004 May 11;218.
BACKGROUND: To empirically evaluate bias in estimation of accuracy associated with delay in verification of diagnosis among studies evaluating tests for predicting endometrial hyperplasia. METHODS: Systematic reviews of all published research on accuracy of miniature endometrial biopsy and endometrial ultrasonography for diagnosing endometrial hyperplasia identified 27 test accuracy studies (2,982 subjects). Of these, 16 had immediate histological verification of diagnosis while 11 had verification delayed > 24 hrs after testing. The effect of delay in verification of diagnosis on estimates of accuracy was evaluated using meta-regression with diagnostic odds ratio (dOR) as the accuracy measure. This analysis was adjusted for study quality and type of test (miniature endometrial biopsy or endometrial ultrasound). RESULTS: Compared to studies with immediate verification of diagnosis (dOR 67.2, 95% CI 21.7-208.8), those with delayed verification (dOR 16.2, 95% CI 8.6-30.5) underestimated the diagnostic accuracy by 74% (95% CI 7%-99%; P value = 0.048). CONCLUSION: Among studies of miniature endometrial biopsy and endometrial ultrasound, diagnostic accuracy is considerably underestimated if there is a delay in histological verification of diagnosis. [Abstract/Link to Full Text]

Andrieu JM, Lu W
Long-term clinical, immunologic and virologic impact of glucocorticoids on the chronic phase of HIV infection.
BMC Med. 2004 May 5;217.
BACKGROUND: To test the hypothesis of down-regulating the increased immune system activation/destruction process associated with chronic HIV infection, we focused our interest on prednisolone (PDN), because we had showed that, in vitro, PDN had a strong anti-apoptotic activity on activated T cells of HIV-infected patients and no effect on viral replication. We thus designed in 1992 a pilot study to evaluate the clinical, immunologic and virologic effects of PDN. The drug was given to a group of 44 patients with CD4 T cells over 200/microl. After one year, no patient had developed clinical AIDS and the mean CD4 T cell count of the group had increased from 441 +/- 21 cells/microl to 553 +/- 43 cells/microl. Moreover, markers of immune activation had dropped back to normal levels while the mean viral load of the group had remained unchanged. Here we explore the long-term clinical, immunologic, and virologic impact of prednisolone on the chronic phase of HIV infection. METHODS: Retrospective study over 10 years starting between July 1992 and February 1993. A total of 44 patients with CD4 cells/microl ranging from 207 to 775 were treated with prednisolone, 0.5 mg/kg/d, over 6 months and 0.3 mg/kg/d thereafter. RESULTS: No clinical AIDS developed under prednisolone; side effects of the drug were mild. CD4 cells which increased from 421 cells/microl at entry to 625 cells/microl at day 15, slowly decreased to reach 426 cells/microl after two years; T cell apoptosis and activation markers dropped within 15 days to normal levels and reincreased slowly thereafter. Serum viral loads remained stable. The percentage of patients maintaining CD4 cells over entry was 43.2% at two years, 11.4% at five years and 4.6% at 10 years. Initial viral load was highly predictive of the rate of CD4 decrease under prednisolone. CONCLUSIONS: Prednisolone postponed CD4 cell decrease in a viral load dependent manner for a median of two years and for up to 10 years in a fraction of the patients with a low viral load. These findings might stimulate clinical trials as well as biological research on the role of antiapoptotic drugs in HIV infection. [Abstract/Link to Full Text]

Lalloo AK, Luo FR, Guo A, Paranjpe PV, Lee SH, Vyas V, Rubin E, Sinko PJ
Membrane transport of camptothecin: facilitation by human P-glycoprotein (ABCB1) and multidrug resistance protein 2 (ABCC2).
BMC Med. 2004 May 4;216.
BACKGROUND: The purpose of the present study was to continue the investigation of the membrane transport mechanisms of 20-(S)-camptothecin (CPT) in order to understand the possible role of membrane transporters on its oral bioavailability and disposition. METHODS: The intestinal transport kinetics of CPT were characterized using Caco-2 cells, MDCKII wild-type cells and MDCKII cells transfected with human P-glycoprotein (PGP) (ABCB1) or human multidrug resistance protein 2 (MRP2) (ABCC2). The effects of drug concentration, inhibitors and temperature on CPT directional permeability were determined. RESULTS: The absorptive (apical to basolateral) and secretory (basolateral to apical) permeabilities of CPT were found to be saturable. Reduced secretory CPT permeabilities with decreasing temperatures suggests the involvement of an active, transporter-mediated secretory pathway. In the presence of etoposide, the CPT secretory permeability decreased 25.6%. However, inhibition was greater in the presence of PGP and of the breast cancer resistant protein inhibitor, GF120918 (52.5%). The involvement of additional secretory transporters was suggested since the basolateral to apical permeability of CPT was not further reduced in the presence of increasing concentrations of GF120918. To investigate the involvement of specific apically-located secretory membrane transporters, CPT transport studies were conducted using MDCKII/PGP cells and MDCKII/MRP2 cells. CPT carrier-mediated permeability was approximately twofold greater in MDCKII/PGP cells and MDCKII/MRP2 cells than in MDCKII/wild-type cells, while the apparent Km values were comparable in all three cell lines. The efflux ratio of CPT in MDCKII/PGP in the presence of 0.2 microM GF120918 was not completely reversed (3.36 to 1.49). However, the decrease in the efflux ratio of CPT in MDCKII/MRP2 cells (2.31 to 1.03) suggests that CPT efflux was completely inhibited by MK571, a potent inhibitor of the Multidrug Resistance Protein transporter family. CONCLUSIONS: The current results provide evidence that PGP and MRP2 mediate the secretory transport of CPT in vitro. However, the involvement of other transporters cannot be ruled out based on these studies. Since these transporters are expressed in the intestine, liver and kidney variations in their expression levels and/or regulation may be responsible for the erratic oral absorption and biliary excretion of CPT observed in human subjects. [Abstract/Link to Full Text]

McGrath J, Saha S, Welham J, El Saadi O, MacCauley C, Chant D
A systematic review of the incidence of schizophrenia: the distribution of rates and the influence of sex, urbanicity, migrant status and methodology.
BMC Med. 2004 Apr 28;213.
BACKGROUND: Understanding variations in the incidence of schizophrenia is a crucial step in unravelling the aetiology of this group of disorders. The aims of this review are to systematically identify studies related to the incidence of schizophrenia, to describe the key features of these studies, and to explore the distribution of rates derived from these studies. METHODS: Studies with original data related to the incidence of schizophrenia (published 1965-2001) were identified via searching electronic databases, reviewing citations and writing to authors. These studies were divided into core studies, migrant studies, cohort studies and studies based on Other Special Groups. Between- and within-study filters were applied in order to identify discrete rates. Cumulative plots of these rates were made and these distributions were compared when the underlying rates were sorted according to sex, urbanicity, migrant status and various methodological features. RESULTS: We identified 100 core studies, 24 migrant studies, 23 cohort studies and 14 studies based on Other Special Groups. These studies, which were drawn from 33 countries, generated a total of 1,458 rates. Based on discrete core data for persons (55 studies and 170 rates), the distribution of rates was asymmetric and had a median value (10%-90% quantile) of 15.2 (7.7-43.0) per 100,000. The distribution of rates was significantly higher in males compared to females; the male/female rate ratio median (10%-90% quantile) was 1.40 (0.9-2.4). Those studies conducted in urban versus mixed urban-rural catchment areas generated significantly higher rate distributions. The distribution of rates in migrants was significantly higher compared to native-born; the migrant/native-born rate ratio median (10%-90% quantile) was 4.6 (1.0-12.8). Apart from the finding that older studies reported higher rates, other study features were not associated with significantly different rate distributions (e.g. overall quality, methods related to case finding, diagnostic confirmation and criteria, the use of age-standardization and age range). CONCLUSIONS: There is a wealth of data available on the incidence of schizophrenia. The width and skew of the rate distribution, and the significant impact of sex, urbanicity and migrant status on these distributions, indicate substantial variations in the incidence of schizophrenia. [Abstract/Link to Full Text]

Recent Articles in BMC Complementary and Alternative Medicine

Caulfield T, DeBow S
A systematic review of how homeopathy is represented in conventional and CAM peer reviewed journals.
BMC Complement Altern Med. 2005;512.
BACKGROUND: Growing popularity of complementary and alternative medicine (CAM) in the public sector is reflected in the scientific community by an increased number of research articles assessing its therapeutic effects. Some suggest that publication biases occur in mainstream medicine, and may also occur in CAM. Homeopathy is one of the most widespread and most controversial forms of CAM. The purpose of this study was to compare the representation of homeopathic clinical trials published in traditional science and CAM journals. METHODS: Literature searches were performed using Medline (PubMed), AMED and Embase computer databases. Search terms included "homeo-pathy, -path, and -pathic" and "clinical" and "trial". All articles published in English over the past 10 years were included. Our search yielded 251 articles overall, of which 46 systematically examined the efficacy of homeopathic treatment. We categorized the overall results of each paper as having either "positive" or "negative" outcomes depending upon the reported effects of homeopathy. We also examined and compared 15 meta-analyses and review articles on homeopathy to ensure our collection of clinical trials was reasonably comprehensive. These articles were found by inserting the term "review" instead of "clinical" and "trial". RESULTS: Forty-six peer-reviewed articles published in a total of 23 different journals were compared (26 in CAM journals and 20 in conventional journals). Of those in conventional journals, 69% reported negative findings compared to only 30% in CAM journals. Very few articles were found to be presented in a "negative" tone, and most were presented using "neutral" or unbiased language. CONCLUSION: A considerable difference exists between the number of clinical trials showing positive results published in CAM journals compared with traditional journals. We found only 30% of those articles published in CAM journals presented negative findings, whereas over twice that amount were published in traditional journals. These results suggest a publication bias against homeopathy exists in mainstream journals. Conversely, the same type of publication bias does not appear to exist between review and meta-analysis articles published in the two types of journals. [Abstract/Link to Full Text]

Sherman KJ, Cherkin DC, Kahn J, Erro J, Hrbek A, Deyo RA, Eisenberg DM
A survey of training and practice patterns of massage therapists in two US states.
BMC Complement Altern Med. 2005;513.
BACKGROUND: Despite the growing popularity of therapeutic massage in the US, little is known about the training or practice characteristics of massage therapists. The objective of this study was to describe these characteristics. METHODS: As part of a study of random samples of complementary and alternative medicine (CAM) practitioners, we interviewed 226 massage therapists licensed in Connecticut and Washington state by telephone in 1998 and 1999 (85% of those contacted) and then asked a sample of them to record information on 20 consecutive visits to their practices (total of 2005 consecutive visits). RESULTS: Most massage therapists were women (85%), white (95%), and had completed some continuing education training (79% in Connecticut and 52% in Washington). They treated a limited number of conditions, most commonly musculoskeletal (59% and 63%) (especially back, neck, and shoulder problems), wellness care (20% and 19%), and psychological complaints (9% and 6%) (especially anxiety and depression). Practitioners commonly used one or more assessment techniques (67% and 74%) and gave a massage emphasizing Swedish (81% and 77%), deep tissue (63% and 65%), and trigger/pressure point techniques (52% and 46%). Self-care recommendations, including increasing water intake, body awareness, and specific forms of movement, were made as part of more than 80% of visits. Although most patients self-referred to massage, more than one-quarter were receiving concomitant care for the same problem from a physician. Massage therapists rarely communicated with these physicians. CONCLUSION: This study provides new information about licensed massage therapists that should be useful to physicians and other healthcare providers interested in learning about massage therapy in order to advise their patients about this popular CAM therapy. [Abstract/Link to Full Text]

Herman PM, Craig BM, Caspi O
Is complementary and alternative medicine (CAM) cost-effective? A systematic review.
BMC Complement Altern Med. 2005;511.
BACKGROUND: Out-of-pocket expenditures of over 34 billion dollars per year in the US are an apparent testament to a widely held belief that complementary and alternative medicine (CAM) therapies have benefits that outweigh their costs. However, regardless of public opinion, there is often little more than anecdotal evidence on the health and economic implications of CAM therapies. The objectives of this study are to present an overview of economic evaluation and to expand upon a previous review to examine the current scope and quality of CAM economic evaluations. METHODS: The data sources used were Medline, AMED, Alt-HealthWatch, and the Complementary and Alternative Medicine Citation Index; January 1999 to October 2004. Papers that reported original data on specific CAM therapies from any form of standard economic analysis were included. Full economic evaluations were subjected to two types of quality review. The first was a 35-item checklist for reporting quality, and the second was a set of four criteria for study quality (randomization, prospective collection of economic data, comparison to usual care, and no blinding). RESULTS: A total of 56 economic evaluations (39 full evaluations) of CAM were found covering a range of therapies applied to a variety of conditions. The reporting quality of the full evaluations was poor for certain items, but was comparable to the quality found by systematic reviews of economic evaluations in conventional medicine. Regarding study quality, 14 (36%) studies were found to meet all four criteria. These exemplary studies indicate CAM therapies that may be considered cost-effective compared to usual care for various conditions: acupuncture for migraine, manual therapy for neck pain, spa therapy for Parkinson's, self-administered stress management for cancer patients undergoing chemotherapy, pre- and post-operative oral nutritional supplementation for lower gastrointestinal tract surgery, biofeedback for patients with "functional" disorders (eg, irritable bowel syndrome), and guided imagery, relaxation therapy, and potassium-rich diet for cardiac patients. CONCLUSION: Whereas the number and quality of economic evaluations of CAM have increased in recent years and more CAM therapies have been shown to be of good value, the majority of CAM therapies still remain to be evaluated. [Abstract/Link to Full Text]

Ahn AC, Wu J, Badger GJ, Hammerschlag R, Langevin HM
Electrical impedance along connective tissue planes associated with acupuncture meridians.
BMC Complement Altern Med. 2005;510.
BACKGROUND: Acupuncture points and meridians are commonly believed to possess unique electrical properties. The experimental support for this claim is limited given the technical and methodological shortcomings of prior studies. Recent studies indicate a correspondence between acupuncture meridians and connective tissue planes. We hypothesized that segments of acupuncture meridians that are associated with loose connective tissue planes (between muscles or between muscle and bone) visible by ultrasound have greater electrical conductance (less electrical impedance) than non-meridian, parallel control segments. METHODS: We used a four-electrode method to measure the electrical impedance along segments of the Pericardium and Spleen meridians and corresponding parallel control segments in 23 human subjects. Meridian segments were determined by palpation and proportional measurements. Connective tissue planes underlying those segments were imaged with an ultrasound scanner. Along each meridian segment, four gold-plated needles were inserted along a straight line and used as electrodes. A parallel series of four control needles were placed 0.8 cm medial to the meridian needles. For each set of four needles, a 3.3 kHz alternating (AC) constant amplitude current was introduced at three different amplitudes (20, 40, and 80 microAmps) to the outer two needles, while the voltage was measured between the inner two needles. Tissue impedance between the two inner needles was calculated based on Ohm's law (ratio of voltage to current intensity). RESULTS: At the Pericardium location, mean tissue impedance was significantly lower at meridian segments (70.4 +/- 5.7 Omega) compared with control segments (75.0 +/- 5.9 Omega) (p = 0.0003). At the Spleen location, mean impedance for meridian (67.8 +/- 6.8 Omega) and control segments (68.5 +/- 7.5 Omega) were not significantly different (p = 0.70). CONCLUSION: Tissue impedance was on average lower along the Pericardium meridian, but not along the Spleen meridian, compared with their respective controls. Ultrasound imaging of meridian and control segments suggested that contact of the needle with connective tissue may explain the decrease in electrical impedance noted at the Pericardium meridian. Further studies are needed to determine whether tissue impedance is lower in (1) connective tissue in general compared with muscle and (2) meridian-associated vs. non meridian-associated connective tissue. [Abstract/Link to Full Text]

Han LK, Zheng YN, Yoshikawa M, Okuda H, Kimura Y
Anti-obesity effects of chikusetsusaponins isolated from Panax japonicus rhizomes.
BMC Complement Altern Med. 2005;59.
BACKGROUND: The rhizomes of Panax japonicus are used as a folk medicine for treatment of life-style related diseases such as arteriosclerosis, hyperlipidemia, hypertension and non-insulin-dependent diabetes mellitus as a substitute for ginseng roots in China and Japan. Obesity is closely associated with life-style-related diseases. This study was performed to clarify whether chikusetsusaponins prevent obesity induced in mice by a high-fat diet for 9 weeks. METHODS: We performed two in vivo experiments. In one, female ICR mice were fed a high-fat diet with or without 1 or 3% chikusetsusaponins isolated from P. japonicus rhizomes for 9 weeks. In the other, lipid emulsion with or without chikusetsusaponins was administered orally to male Wistar rats, and then the plasma triacylglycerol level was measured 0.5 to 5 h after the orally administered lipid emulsion. For in vitro experiments, the inhibitory effects of total chikusetsusaponins and various purified chikusetsusaponins on pancreatic lipase activity were determined by measuring the rate of release of oleic acid from triolein in an assay system using triolein emulsified with lecithin. RESULTS: Total chikusetsusaponins prevented the increases in body weight and parametrial adipose tissue weight induced by a high-fat diet. Furthermore, consumption of a high-fat diet containing 1 or 3% total chikusetsusaponins significantly increased the fecal content and triacylglycerol level at day 3 compared with the high-fat diet groups. Total chikusetsusaponins inhibited the elevation of the plasma triacylglycerol level 2 h after the oral administration of the lipid emulsion. Total chikusetsusaponins, chikusetsusaponin III, 28-deglucosyl-chikusetsusaponin IV and 28-deglucosyl-chikusetsusaponin V inhibited the pancreatic lipase activity. CONCLUSION: The anti-obesity effects of chikusetsusaponins isolated from P. japonicus rhizomes in mice fed a high-fat diet may be partly mediated through delaying the intestinal absorption of dietary fat by inhibiting pancreatic lipase activity. The present study clearly indicated that the saponin fractions of P. japonicus rhizomes had a significant anti-obesity action and supports the traditional usage as a substitute drug for ginseng roots. [Abstract/Link to Full Text]

Inaba R, Mirbod SM, Sugiura H
Effects of Maharishi Amrit Kalash 5 as an Ayurvedic herbal food supplement on immune functions in aged mice.
BMC Complement Altern Med. 2005;58.
BACKGROUND: Maharishi Amrit Kalash (MAK) 5, one of the Ayurvedic food supplements, belongs to a group of substances known as Rasayana. MAK5 and other Rasayanas are believed to enhance the body's resistance to infections and disease, and enhance longevity. In this study, we determined the effects of administration of MAK5, one of the Ayurvedic food supplements on immune functions in young and old mice. METHODS: Male C3H/He N mice were divided into five groups: two no treatment groups (old control: 22-month-old and young control: 2-month-old) and three MAK5 treated groups with differing dose of MAK5. MAK5 was given p.o. at 50 mg/kg, 100 mg/kg or 200 mg/kg per day (3 days/week) for 2 months. RESULTS: We found that glucose consumption of peritoneal macrophages from old mice treated with MAK5 at all doses and incubated for 48 and 72 h were significantly greater than that in the control group. Nitric oxide production of peritoneal macrophages stimulated by lipopolysaccharide (LPS) in old mice treated with MAK5 at all doses was significantly greater than that in the old control group, but not compared to the young control group. Stimulation index (S.I.) in old mice gavaged with MAK5 at all doses was significantly higher than that in the old control group. IL-2 production stimulated by Con A in old mice given MAK5 at all doses was significantly higher than that in the old control group. Production of IFN-gamma stimulated by Con A in old mice given MAK5 at doses of 100 mg/kg and 200 mg/kg were significantly higher than that in the old control group. IL-4 production of splenic lymphocyte stimulated by Con A in old mice given MAK5 at dose levels of 100 and 200 mg/kg were significantly higher than that in the old control group. CONCLUSION: The results suggest that MAK5 suppressed the age associated glucose consumption of peritoneal macrophages and cellular immune function reduction, and that it contributes to the prevention of the immunosenescence. [Abstract/Link to Full Text]

Akinyemi KO, Oladapo O, Okwara CE, Ibe CC, Fasure KA
Screening of crude extracts of six medicinal plants used in South-West Nigerian unorthodox medicine for anti-methicillin resistant Staphylococcus aureus activity.
BMC Complement Altern Med. 2005;56.
BACKGROUND: Six Nigerian medicinal plants Terminalia avicennioides, Phylantus discoideus, Bridella ferruginea, Ageratum conyzoides, Ocimum gratissimum and Acalypha wilkesiana used by traditional medical practitioners for the treatment of several ailments of microbial and non-microbial origins were investigated for in vitro anti-methicillin Resistant Staphylococcus aureus (MRSA) activity. METHODS: Fresh plant materials were collected from the users. Water and ethanol extracts of the shredded plants were obtained by standard methods. The Bacterial cultures used were strains of MRSA isolated from patients. MRSA was determined by the reference broth microdilution methods using the established National Committee for Clinical Laboratory Standards break points. Staphylococcus aureus NCIB 8588 was used as a standard strain. Susceptibility testing and phytochemical screening of the plant extracts were performed by standard procedures. Controls were maintained for each test batch. RESULTS: Both water and ethanol extracts of T. avicennioides, P. discoideus, O. gratissimum, and A. wilkesiana were effective on MRSA. The Minimum Inhibition Concentration (MIC) and Minimum Bactericidal Concentration (MBC) of the ethanol extracts of these plants range from 18.2 to 24.0 mcg/ml and 30.4 to 37.0 mcg/ml respectively. In contrast, MIC range of 30.6 to 43.0 mcg/ml and 55.4 to 71.0 mcg/ml were recorded for ethanol and water extracts of B. ferruginea, and A. conyzoides respectively. Higher MBC values were obtained for the two plants. These concentrations were too high to be considered active in this study. All the four active plants contained at least trace amount of anthraquinones. CONCLUSION: Our results offer a scientific basis for the traditional use of water and ethanol extracts of A. wilkesiana, O. gratissimum, T. avicennioides and P. discoideus against MRSA-associated diseases. However, B. ferruginea and A. conyzoides were ineffective in vitro in this study; we therefore suggest the immediate stoppage of their traditional use against MRSA-associated diseases in Lagos, Nigeria. [Abstract/Link to Full Text]

Samoszuk M, Tan J, Chorn G
The chalcone butein from Rhus verniciflua Stokes inhibits clonogenic growth of human breast cancer cells co-cultured with fibroblasts.
BMC Complement Altern Med. 2005;55.
BACKGROUND: Butein (3,4,2',4'-tetrahydroxychalone), a plant polyphenol, is a major biologically active component of the stems of Rhus verniciflua Stokes. It has long been used as a food additive in Korea and as an herbal medicine throughout Asia. Recently, butein has been shown to suppress the functions of fibroblasts. Because fibroblasts are believed to play an important role in promoting the growth of breast cancer cells, we investigated the ability of butein to inhibit the clonogenic growth of small numbers of breast cancer cells co-cultured with fibroblasts in vitro. METHODS: We first measured the clonogenic growth of small numbers of the UACC-812 human breast cancer cell line co-cultured on monolayers of serum-activated, human fibroblasts in the presence of butein (2 microg/mL) or various other modulators of fibroblast function (troglitazone-1 microg/mL; GW9662-1 microM; meloxican-1 microM; and 3,4 dehydroproline-10 microg/mL). In a subsequent experiment, we measured the dose-response effect on the clonogenic growth of UACC-812 breast cancer cells by pre-incubating the fibroblasts with varying concentrations of butein (10 microg/ml-1.25 microg/mL). Finally, we measured the clonogenic growth of primary breast cancer cells obtained from 5 clinical specimens with normal fibroblasts and with fibroblasts that had been pre-treated with a fixed dose of butein (2.5 microg/mL). RESULTS: Of the five modulators of fibroblast function that we tested, butein was by far the most potent inhibitor of clonogenic growth of UACC-812 breast cancer cells co-cultured with fibroblasts. Pre-treatment of fibroblasts with concentrations of butein as low as 2.5 microg/mL nearly abolished subsequent clonogenic growth of UACC-812 breast cancer cells co-cultured with the fibroblasts. A similar dose of butein had no effect on the clonogenic growth of breast cancer cells cultured in the absence of fibroblasts. Significantly, clonogenic growth of the primary breast cancer cells was also significantly reduced or abolished when the tumor cells were co-cultured with fibroblasts that had been pre-treated with a fixed dose of butein. CONCLUSION: We conclude that fibroblasts pre-treated with non-toxic doses of butein (a natural herbal compound) no longer support the clonogenic growth of small numbers of primary breast cancer cells seeded into co-cultures. These results suggest that interference with the interaction between fibroblasts and breast cancer cells by the natural herbal compound, butein, should be further investigated as a novel experimental approach for possibly suppressing the growth of micrometastases of breast cancer. [Abstract/Link to Full Text]

Chagan L, Bernstein D, Cheng JW, Kirschenbaum HL, Rozenfeld V, Caliendo GC, Meyer J, Mehl B
Use of biological based therapy in patients with cardiovascular diseases in a university-hospital in New York City.
BMC Complement Altern Med. 2005;54.
BACKGROUND: The use of complementary and alternative products including Biological Based Therapy (BBT) has increased among patients with various medical illnesses and conditions. The studies assessing the prevalence of BBT use among patients with cardiovascular diseases are limited. Therefore, an evaluation of BBT in this patient population would be beneficial. This was a survey designed to determine the effects of demographics on the use of Biological Based Therapy (BBT) in patients with cardiovascular diseases. The objective of this study was to determine the effect of the education level on the use of BBT in cardiovascular patients. This survey also assessed the perceptions of users regarding the safety/efficacy of BBT, types of BBT used and potential BBT-drug interactions. METHOD: The survey instrument was designed to assess the findings. Patients were interviewed from February 2001 to December 2002. 198 inpatients with cardiovascular diseases (94 BBT users and 104 non-users) in a university hospital were included in the study. RESULTS: Users had a significantly higher level of education than non-users (college graduate: 28 [30%] versus 12 [12%], p = 0.003). Top 10 BBT products used were vitamin E [41(43.6%)], vitamin C [30(31.9%)], multivitamins [24(25.5%)], calcium [19(20.2%)], vitamin B complex [17(18.1%)], fish oil [12(12.8%)], coenzyme Q10 [11(11.7%)], glucosamine [10(10.6%)], magnesium [8(8.5%)] and vitamin D [6(6.4%)]. Sixty percent of users' physicians knew of the BBT use. Compared to non-users, users believed BBT to be safer (p < 0.001) and more effective (p < 0.001) than prescription drugs. Forty-two potential drug-BBT interactions were identified. CONCLUSION: Incidence of use of BBT in cardiovascular patients is high (47.5%), as is the risk of potential drug interaction. Health care providers need to monitor BBT use in patients with cardiovascular diseases. [Abstract/Link to Full Text]

Clement YN, Williams AF, Aranda D, Chase R, Watson N, Mohammed R, Stubbs O, Williamson D
Medicinal herb use among asthmatic patients attending a specialty care facility in Trinidad.
BMC Complement Altern Med. 2005 Feb 15;53.
BACKGROUND: There is an increasing prevalence of asthma in the Caribbean and patients remain non-compliant to therapy despite the development of guidelines for management and prevention. Some patients may self-medicate with medicinal herbs for symptomatic relief, as there is a long tradition of use for a variety of ailments. The study assessed the prevalence of use and the factors affecting the decision to use herbs in asthmatic patients attending a public specialty care clinic in Trinidad. METHODS: A descriptive, cross-sectional study was conducted at the Chest Clinic in Trinidad using a de novo, pilot-tested, researcher-administered questionnaire between June and July 2003. RESULTS: Fifty-eight out of 191 patients (30.4%) reported using herbal remedies for symptomatic relief. Gender, age, ethnicity, and asthma severity did not influence the decision to use herbs; however, 62.5% of patients with tertiary level schooling used herbs, p = 0.025. Thirty-four of these 58 patients (58.6%) obtained herbs from their backyards or the supermarket; only 14 patients (24.1%) obtained herbs from an herbalist, herbal shop or pharmacy. Relatives and friends were the sole source of information for most patients (70.7%), and only 10.3% consulted an herbalist. Ginger, garlic, aloes, shandileer, wild onion, pepper and black sage were the most commonly used herbs. CONCLUSIONS: Among patients attending the Chest Clinic in Trinidad the use of herbal remedies in asthma is relatively common on the advice of relatives and friends. It is therefore becoming imperative for healthcare providers to become more knowledgeable on this modality and to keep abreast with the latest developments. [Abstract/Link to Full Text]

Taft R, Moore D, Yount G
Time-lapse analysis of potential cellular responsiveness to Johrei, a Japanese healing technique.
BMC Complement Altern Med. 2005 Jan 24;52.
BACKGROUND: Johrei is an alternative healing practice which involves the channeling of a purported universal healing energy to influence the health of another person. Despite little evidence to support the efficacy of such practices the use of such treatments is on the rise. METHODS: We assessed cultured human cancer cells for potential responsiveness to Johrei treatment from a short distance. Johrei treatment was delivered by practitioners who participated in teams of two, alternating every half hour for a total of four hours of treatment. The practitioners followed a defined set of mental procedures to minimize variability in mental states between experiments. An environmental chamber maintained optimal growth conditions for cells throughout the experiments. Computerized time-lapse microscopy allowed documentation of cancer cell proliferation and cell death before, during and after Johrei treatments. RESULTS: Comparing eight control experiments with eight Johrei intervention experiments, we found no evidence of a reproducible cellular response to Johrei treatment. CONCLUSION: Cell death and proliferation rates of cultured human cancer cells do not appear responsive to Johrei treatment from a short distance. [Abstract/Link to Full Text]

Sreemantula S, Nammi S, Kolanukonda R, Koppula S, Boini KM
Adaptogenic and nootropic activities of aqueous extract of Vitis vinifera (grape seed): an experimental study in rat model.
BMC Complement Altern Med. 2005 Jan 19;51.
BACKGROUND: The aerial parts of Vitis vinifera (common grape or European grape) have been widely used in Ayurveda to treat a variety of common and stress related disorders. In the present investigation, the seed extract of V. vinifera was evaluated for antistress activity in normal and stress induced rats. Furthermore, the extract was studied for nootropic activity in rats and in-vitro antioxidant potential to correlate its antistress activity. METHODS: For the evaluation of antistress activity, groups of rats (n = 6) were subjected to forced swim stress one hour after daily treatment of V. vinifera extract. Urinary vanillylmandelic acid (VMA) and ascorbic acid were selected as non-invasive biomarkers to assess the antistress activity. The 24 h urinary excretion of vanillylmandelic acid (VMA) and ascorbic acid were determined by spectrophotometric methods in all groups under normal and stressed conditions. The nootropic activity of the extract as determined from acquisition, retention and retrieval in rats was studied by conditioned avoidance response using Cook's pole climbing apparatus. The in vitro antioxidant activity was determined based on the ability of V. vinifera to scavenge hydroxyl radicals. RESULTS: Daily administration of V. vinifera at doses of 100, 200 and 300 mg/kg body weight one hour prior to induction of stress inhibited the stress induced urinary biochemical changes in a dose dependent manner. However, no change in the urinary excretion of VMA and ascorbic acid was observed in normal animals at all the doses studied. The cognition, as determined by the acquisition, retention and recovery in rats was observed to be dose dependent. The extract also produced significant inhibition of hydroxyl radicals in comparison to ascorbic acid in a dose dependent manner. CONCLUSION: The present study provides scientific support for the antistress (adaptogenic), antioxidant and nootropic activities of V. vinifera seed extract and substantiate the traditional claims for the usage of grape fruits and seeds in stress induced disorders. [Abstract/Link to Full Text]

Teekachunhatean S, Kunanusorn P, Rojanasthien N, Sananpanich K, Pojchamarnwiputh S, Lhieochaiphunt S, Pruksakorn S
Chinese herbal recipe versus diclofenac in symptomatic treatment of osteoarthritis of the knee: a randomized controlled trial [ISRCTN70292892].
BMC Complement Altern Med. 2004 Dec 13;419.
BACKGROUND: Duhuo Jisheng Wan (DJW) is perhaps the best known and most widely used Chinese herbal recipe for arthralgia, but the clinical study to verify its efficacy is lacking. The purpose of this study was to compare the efficacy of DJW versus diclofenac in symptomatic treatment of osteoarthritis (OA) of the knee. METHODS: This study was a randomized, double-blind, double-dummy, controlled trial. The 200 patients suffering from OA of the knee, were randomized into the DJW and diclofenac group. The patients were evaluated after a run-in period of one week (week 0) and then weekly during 4 weeks of treatment. The clinical assessments included visual analog scale (VAS) score that assessed pain and stiffness, Lequesne's functional index, time for climbing up 10 steps, as well as physician's and patients' overall opinions on improvement. RESULTS: Ninety four patients in each group completed the study. In the first few weeks of treatment, the mean changes in some variables (VAS, which assessed walking pain, standing pain and stiffness, as well as Lequesne's functional index) of the DJW group were significantly lower than those of the diclofenac group. Afterwards, these mean changes became no different throughout the study. Most of the physician's and patients' overall opinions on improvement at each time point did not significantly differ between the two groups. Approximately 30% of patients in both groups experienced mild adverse events. CONCLUSION: DJW demonstrates clinically comparable efficacy to diclofenac after 4 weeks of treatment. However, the slow onset of action as well as approximately equal rate of adverse events to diclofenac might limit its alternative role in treatment of OA of the knee. [Abstract/Link to Full Text]

Kuo GM, Hawley ST, Weiss LT, Balkrishnan R, Volk RJ
Factors associated with herbal use among urban multiethnic primary care patients: a cross-sectional survey.
BMC Complement Altern Med. 2004 Dec 2;418.
BACKGROUND: The use of herbal supplements in the United States has become increasingly popular. The prevalence of herbal use among primary care patients varies in previous studies; the pattern of herbal use among urban racially/ethnically diverse primary care patients has not been widely studied. The primary objectives of this study were to describe the use of herbs by ethnically diverse primary care patients in a large metropolitan area and to examine factors associated with such use. The secondary objective was to investigate perceptions about and patterns of herbal use. METHODS: Data for a cross-sectional survey were collected at primary care practices affiliated with the Southern Primary-care Urban Research Network (SPUR-Net) in Houston, Texas, from September 2002 to March 2003. To participate in the study, patients had to be at least 18 years of age and visiting one of the SPUR-Net clinics for routine, nonacute care. Survey questions were available in both English and Spanish. RESULTS: A total of 322 patients who had complete information on race/ethnicity were included in the analysis. Overall, 36% of the surveyed patients (n = 322) indicated use of herbs, with wide variability among ethnic groups: 50% of Hispanics, 50% of Asians, 41% of Whites, and 22% of African-Americans. Significant factors associated with an individual's herbal use were ethnicity other than African-American, having an immigrant family history, and reporting herbal use by other family members. About 40% of survey respondents believed that taking prescription medications and herbal medicines together was more effective than taking either alone. One-third of herbal users reported using herbs on a daily basis. More Whites (67%) disclosed their herbal use to their health-care providers than did African-Americans (45%), Hispanics (31%), or Asians (31%). CONCLUSIONS: Racial/ethnic differences in herbal use were apparent among this sample of urban multiethnic adult primary care patients. Associated factors of herbal use were non-African-American ethnicity, immigrant family history, and herbal use among family members. Whereas Hispanics and Asians reported the highest rates of herbal use, they were the least likely to disclose their use to health-care professionals. These findings are important for ensuring medication safety in primary care practices. [Abstract/Link to Full Text]

Tajuddin S, Latif A, Qasmi IA
Effect of 50% ethanolic extract of Syzygium aromaticum (L.) Merr. & Perry. (clove) on sexual behaviour of normal male rats.
BMC Complement Altern Med. 2004 Nov 5;417.
BACKGROUND: The flower bud of Syzygium aromaticum (L.) Merr. & Perry. (clove) has been used in Unani medicine since ancient times for the treatment of male sexual disorders. The present study is aimed to investigate the effect of 50% ethanolic extract of clove on general mating behaviour, libido, potency along with its likely gastric ulceration and adverse effects on sexually normal male albino rats. METHODS: The suspension of the extract was administered orally at the dose of 100, 250, and 500 mg / kg, to different groups of male rats (n = 6) once a day for seven days. The female albino rats involved in mating were made receptive by hormonal treatment. The general mating behaviour, libido and potency were determined and compared with the standard reference drug sildenafil citrate. The probable gastric ulceration and adverse effects of the extract were also evaluated. RESULTS: Oral administration of the extract significantly increased the Mounting Frequency, Intromission Frequency; Intromission Latency, Erections; Quick Flips, Long Flips as well as aggregate of penile reflexes and caused significant reduction in the Mounting Latency and Post Ejaculatory Interval. The most appreciable effect of the extract was observed at the dose of 500 mg/kg. The test drug was also found to be devoid of any conspicuous gastric ulceration and adverse effects. CONCLUSION: The results indicated that the 50% ethanolic extract of clove produced a significant and sustained increase in the sexual activity of normal male rats, without any conspicuous gastric ulceration and adverse effects. Thus, the resultant aphrodisiac effectivity of the extract lends support to the claims for its traditional usage in sexual disorders. [Abstract/Link to Full Text]

Pari L, Latha M
Protective role of Scoparia dulcis plant extract on brain antioxidant status and lipidperoxidation in STZ diabetic male Wistar rats.
BMC Complement Altern Med. 2004 Nov 2;416.
BACKGROUND: The aim of the study was to investigate the effect of aqueous extract of Scoparia dulcis on the occurrence of oxidative stress in the brain of rats during diabetes by measuring the extent of oxidative damage as well as the status of the antioxidant defense system. METHODS: Aqueous extract of Scoparia dulcis plant was administered orally (200 mg/kg body weight) and the effect of extract on blood glucose, plasma insulin and the levels of thiobarbituric acid reactive substances (TBARS), hydroperoxides, superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione-S-transferase (GST) and reduced glutathione (GSH) were estimated in streptozotocin (STZ) induced diabetic rats. Glibenclamide was used as standard reference drug. RESULTS: A significant increase in the activities of plasma insulin, superoxide dismutase, catalase, glutathione peroxidase, glutathione-S-transferase and reduced glutathione was observed in brain on treatment with 200 mg/kg body weight of Scoparia dulcis plant extract (SPEt) and glibenclamide for 6 weeks. Both the treated groups showed significant decrease in TBARS and hydroperoxides formation in brain, suggesting its role in protection against lipidperoxidation induced membrane damage. CONCLUSIONS: Since the study of induction of the antioxidant enzymes is considered to be a reliable marker for evaluating the antiperoxidative efficacy of the medicinal plant, these findings suggest a possible antiperoxidative role for Scoparia dulcis plant extract. Hence, in addition to antidiabetic effect, Scoparia dulcis possess antioxidant potential that may be used for therapeutic purposes. [Abstract/Link to Full Text]

Anick DJ
High sensitivity 1H-NMR spectroscopy of homeopathic remedies made in water.
BMC Complement Altern Med. 2004 Nov 1;415.
BACKGROUND: The efficacy of homeopathy is controversial. Homeopathic remedies are made via iterated shaking and dilution, in ethanol or in water, from a starting substance. Remedies of potency 12 C or higher are ultra-dilute (UD), i.e. contain zero molecules of the starting material. Various hypotheses have been advanced to explain how a UD remedy might be different from unprepared solvent. One such hypothesis posits that a remedy contains stable clusters, i.e. localized regions where one or more hydrogen bonds remain fixed on a long time scale. High sensitivity proton nuclear magnetic resonance spectroscopy has not previously been used to look for evidence of differences between UD remedies and controls. METHODS: Homeopathic remedies made in water were studied via high sensitivity proton nuclear magnetic resonance spectroscopy. A total of 57 remedy samples representing six starting materials and spanning a variety of potencies from 6 C to 10 M were tested along with 46 controls. RESULTS: By presaturating on the water peak, signals could be reliably detected that represented H-containing species at concentrations as low as 5 microM. There were 35 positions where a discrete signal was seen in one or more of the 103 spectra, which should theoretically have been absent from the spectrum of pure water. Of these 35, fifteen were identified as machine-generated artifacts, eight were identified as trace levels of organic contaminants, and twelve were unexplained. Of the unexplained signals, six were seen in just one spectrum each. None of the artifacts or unexplained signals occurred more frequently in remedies than in controls, using a p < .05 cutoff. Some commercially prepared samples were found to contain traces of one or more of these small organic molecules: ethanol, acetate, formate, methanol, and acetone. CONCLUSION: No discrete signals suggesting a difference between remedies and controls were seen, via high sensitivity 1H-NMR spectroscopy. The results failed to support a hypothesis that remedies made in water contain long-lived non-dynamic alterations of the H-bonding pattern of the solvent. [Abstract/Link to Full Text]

Boon HS, Cherkin DC, Erro J, Sherman KJ, Milliman B, Booker J, Cramer EH, Smith MJ, Deyo RA, Eisenberg DM
Practice patterns of naturopathic physicians: results from a random survey of licensed practitioners in two US States.
BMC Complement Altern Med. 2004 Oct 20;414.
BACKGROUND: Despite the growing use of complementary and alternative medicine (CAM) by consumers in the U.S., little is known about the practice of CAM providers. The objective of this study was to describe and compare the practice patterns of naturopathic physicians in Washington State and Connecticut. METHODS: Telephone interviews were conducted with state-wide random samples of licensed naturopathic physicians and data were collected on consecutive patient visits in 1998 and 1999. The main outcome measures were: Sociodemographic, training and practice characteristics of naturopathic physicians; and demographics, reasons for visit, types of treatments, payment source and visit duration for patients. RESULT: One hundred and seventy practitioners were interviewed and 99 recorded data on a total of 1817 patient visits. Naturopathic physicians in Washington and Connecticut had similar demographic and practice characteristics. Both the practitioners and their patients were primarily White and female. Almost 75% of all naturopathic visits were for chronic complaints, most frequently fatigue, headache, and back symptoms. Complete blood counts, serum chemistries, lipids panels and stool analyses were ordered for 4% to 10% of visits. All other diagnostic tests were ordered less frequently. The most commonly prescribed naturopathic therapeutics were: botanical medicines (51% of visits in Connecticut, 43% in Washington), vitamins (41% and 43%), minerals (35% and 39%), homeopathy (29% and 19%) and allergy treatments (11% and 13%). The mean visit length was about 40 minutes. Approximately half the visits were paid directly by the patient. CONCLUSION: This study provides information that will help other health care providers, patients and policy makers better understand the nature of naturopathic care. [Abstract/Link to Full Text]

Gagnier JJ, Chrubasik S, Manheimer E
Harpgophytum procumbens for osteoarthritis and low back pain: a systematic review.
BMC Complement Altern Med. 2004 Sep 15;413.
BACKGROUND: The objective of this review is to determine the effectiveness of Harpagophytum procumbens preparations in the treatment of various forms of musculoskeletal pain. METHODS: Several databases and other sources were searched to identify randomized controlled trials, quasi-randomized controlled trials, and controlled clinical trials testing Harpagophytum preparations in adults suffering from pain due to osteoarthritis or low back pain. RESULTS: Given the clinical heterogeneity and insufficient data for statistical pooling, trials were described in a narrative way, taking into consideration methodological quality scores. Twelve trials were included with six investigating osteoarthritis (two were identical trials), four low back pain, and three mixed-pain conditions. CONCLUSIONS: There is limited evidence for an ethanolic Harpagophytum extract containing less than <30 mg harpagoside per day in the treatment of knee and hip osteoarthritis. There is moderate evidence of effectiveness for (1) the use of a Harpagophytum powder at 60 mg harpagoside in the treatment of osteoarthritis of the spine, hip and knee; (2) the use of an aqueous Harpagophytum extract at a daily dose of 100 mg harpagoside in the treatment of acute exacerbations of chronic non-specific low back pain; and (3) the use of an aqueous extract of Harpagophytum procumbens at 60 mg harpagoside being non-inferior to 12.5 mg rofecoxib per day for chronic non-specific low-back pain (NSLBP) in the short term. Strong evidence exists for the use of an aqueous Harpagophytum extract at a daily dose equivalent of 50 mg harpagoside in the treatment of acute exacerbations of chronic NSLBP. [Abstract/Link to Full Text]

Akhondzadeh S, Fallah-Pour H, Afkham K, Jamshidi AH, Khalighi-Cigaroudi F
Comparison of Crocus sativus L. and imipramine in the treatment of mild to moderate depression: a pilot double-blind randomized trial [ISRCTN45683816].
BMC Complement Altern Med. 2004 Sep 2;412.
BACKGROUND: The morbidity and mortality associated with depression are considerable and continue to increase. Depression currently ranks fourth among the major causes of disability worldwide, after lower respiratory infections, prenatal conditions, and HIV/AIDS. Crocus sativus L. is used to treat depression. Many medicinal plants textbooks refer to this indication whereas there is no evidence-based document. Our objective was to compare the efficacy of stigmas of Crocus sativus (saffron) with imipramine in the treatment of mild to moderate depression in a 6-week pilot double-blind randomized trial. METHODS: Thirty adult outpatients who met the Diagnostic and Statistical Manual of Mental Disorders, 4th edition for major depression based on the structured clinical interview for DSM IV participated in the trial. Patients have a baseline Hamilton Rating Scale for Depression score of at least 18. In this double-blind, single-center trial, patients were randomly assigned to receive capsule of saffron 30 mg/day (TDS) (Group 1) and capsule of imipramine 100 mg/day (TDS) (Group 2) for a 6-week study. RESULTS: Saffron at this dose was found to be effective similar to imipramine in the treatment of mild to moderate depression (F = 2.91, d.f. = 1, P = 0.09). In the imipramine group anticholinergic effects such as dry mouth and also sedation were observed more often that was predictable. CONCLUSION: The main overall finding from this study is that saffron may be of therapeutic benefit in the treatment of mild to moderate depression. To the best of our knowledge this is the first clinical trial that supports this indication for saffron. A large-scale trial with placebo control is warranted. [Abstract/Link to Full Text]

Rawal AK, Muddeshwar MG, Biswas SK
Rubia cordifolia, Fagonia cretica linn and Tinospora cordifolia exert neuroprotection by modulating the antioxidant system in rat hippocampal slices subjected to oxygen glucose deprivation.
BMC Complement Altern Med. 2004 Aug 13;411.
BACKGROUND: The major damaging factor during and after the ischemic/hypoxic insult is the generation of free radicals, which leads to apoptosis, necrosis and ultimately cell death. Rubia cordifolia (RC), Fagonia cretica linn (FC) and Tinospora cordifolia (TC) have been reported to contain a wide variety of antioxidants and have been in use in the eastern system of medicine for various disorders. However, their mechanism of action was largely unknown. We therefore selected these herbs for the present study to test their neuroprotective ability and the associated mechanism in rat hippocampal slices subjected to oxygen-glucose deprivation (OGD). METHODS: Hippocampal Slices were subjected to OGD (oxygen glucose deprivation) and divided into 3 groups: control, OGD and OGD + drug treated. Cytosolic Cu-Zn superoxide dismutase (Cu-Zn SOD), reduced glutathione (GSH), glutathione peroxidase (GPx), nitric oxide (NO) was measured as nitrite (NO2) in the supernatant and protein assays were performed in the respective groups at various time intervals. EPR was used to establish the antioxidant effect of RC, FC and TC with respect to superoxide anion (O2.-), hydroxyl radicals (. OH), nitric oxide (NO) radical and peroxynitrite anion (ONOO) generated from pyrogallol, menadione, DETA-NO and Sin-1 respectively. RT-PCR was performed for the three groups for GCLC, iNOS, Cu-Zn SOD and GAPDH gene expression. RESULTS: All the three herbs were effective in elevating the GSH levels, expression of the gamma-glutamylcysteine ligase and Cu-Zn SOD genes. The herbs also exhibited strong free radical scavenging properties against reactive oxygen and nitrogen species as studied by electron paramagnetic resonance spectroscopy. In addition all the three herbs significantly diminished the expression of iNOS gene after 48 hours which plays a major role in neuronal injury during hypoxia/ischemia. CONCLUSIONS: RC, FC and TC therefore attenuate oxidative stress mediated cell injury during OGD and exert the above effects at both the cytosolic as well as at gene expression level and may be an effective therapeutic tool against ischemic brain damage. [Abstract/Link to Full Text]

Skibola CF
The effect of Fucus vesiculosus, an edible brown seaweed, upon menstrual cycle length and hormonal status in three pre-menopausal women: a case report.
BMC Complement Altern Med. 2004 Aug 4;410.
BACKGROUND: Rates of estrogen-dependent cancers are among the highest in Western countries and lower in the East. These variations may be attributable to differences in dietary exposures such as higher seaweed consumption among Asian populations. The edible brown kelp, Fucus vesiculosus (bladderwrack), as well as other brown kelp species, lower plasma cholesterol levels. Since cholesterol is a precursor to sex hormone biosynthesis, kelp consumption may alter circulating sex hormone levels and menstrual cycling patterns. In particular, dietary kelp may be beneficial to women with or at high risk for estrogen-dependent diseases. To test this, bladderwrack was administered to three pre-menopausal women with abnormal menstrual cycling patterns and/or menstrual-related disease histories. CASE PRESENTATION: Intake of bladderwrack was associated with significant increases in menstrual cycle lengths, ranging from an increase of 5.5 to 14 days. In addition, hormone measurements ascertained for one woman revealed significant anti-estrogenic and progestagenic effects following kelp administration. Mean baseline 17beta-estradiol levels were reduced from 626 +/- 91 to 164 +/- 30 pg/ml (P = 0.04) following 700 mg/d, which decreased further to 92.5.0 +/- 3.5pg/ml (P = 0.03) with the 1.4 g/d dose. Mean baseline progesterone levels rose from 0.58 +/- 0.14 to 8.4 +/- 2.6 ng/ml with the 700 mg/d dose (P = 0.1), which increased further to 16.8 +/- 0.7 ng/ml with the 1.4 g/d dose (P = 0.002). CONCLUSIONS: These pilot data suggest that dietary bladderwrack may prolong the length of the menstrual cycle and exert anti-estrogenic effects in pre-menopausal women. Further, these studies also suggest that seaweed may be another important dietary component apart from soy that is responsible for the reduced risk of estrogen-related cancers observed in Japanese populations. However, these studies will need to be performed in well-controlled clinical trials to confirm these preliminary findings. [Abstract/Link to Full Text]

Sherman KJ, Cherkin DC, Connelly MT, Erro J, Savetsky JB, Davis RB, Eisenberg DM
Complementary and alternative medical therapies for chronic low back pain: What treatments are patients willing to try?
BMC Complement Altern Med. 2004 Jul 19;49.
BACKGROUND: Although back pain is the most common reason patients use complementary and alternative medical (CAM) therapies, little is known about the willingness of primary care back pain patients to try these therapies. As part of an effort to refine recruitment strategies for clinical trials, we sought to determine if back pain patients are willing to try acupuncture, chiropractic, massage, meditation, and t'ai chi and to learn about their knowledge of, experience with, and perceptions about each of these therapies. METHODS: We identified English-speaking patients with diagnoses consistent with chronic low back pain using automated visit data from one health care organization in Boston and another in Seattle. We were able to confirm the eligibility status (i.e., current low back pain that had lasted at least 3 months) of 70% of the patients with such diagnoses and all eligible respondents were interviewed. RESULTS: Except for chiropractic, knowledge about these therapies was low. Chiropractic and massage had been used by the largest fractions of respondents (54% and 38%, respectively), mostly for back pain (45% and 24%, respectively). Among prior users of specific CAM therapies for back pain, massage was rated most helpful. Users of chiropractic reported treatment-related "significant discomfort, pain or harm" more often (23%) than users of other therapies (5-16%). Respondents expected massage would be most helpful (median of 7 on a 0 to 10 scale) and meditation least helpful (median of 3) in relieving their current pain. Most respondents indicated they would be "very likely" to try acupuncture, massage, or chiropractic for their back pain if they did not have to pay out of pocket and their physician thought it was a reasonable treatment option. CONCLUSIONS: Most patients with chronic back pain in our sample were interested in trying therapeutic options that lie outside the conventional medical spectrum. This highlights the need for additional studies evaluating their effectiveness and suggests that researchers conducting clinical trials of these therapies may not have difficulties recruiting patients. [Abstract/Link to Full Text]

Khan MA, Khan NA, Qasmi IA, Ahmad G, Zafar S
Protective effect of Arque-Ajeeb on acute experimental diarrhoea in rats.
BMC Complement Altern Med. 2004 Jul 6;48.
BACKGROUND: Diarrhoea is a major health problem for children worldwide, accounting for 5-8 million deaths each year. Arque-Ajeeb (AA) is a compound formulation of Unani medicine. It is reputed for its beneficial effects in the treatment of diarrhoea and cholera, but the claim of its efficacy is yet to be tested. Therefore the present study has been planned to investigate the real efficacy of this drug in rats. METHODS: The effect of Arque-Ajeeb was investigated for antidiarrhoeal activity against charcoal-induced gut transit, serotonin-induced diarrhoea and PGE2-induced small intestine enteropooling in rats. The control, standard and test groups of experimental animals were administered with normal saline (p.o.), diphenoxylate hydrochloride (5 mg/kg, p.o.) and Arque-Ajeeb (0.07 ml and 0.14 ml/kg, p.o.) respectively except the control group of PGE2-induced small intestine enteropooling which received only 5% ethanol in normal saline (i.p.). Charcoal (10 ml/kg, p.o.) and serotonin (600 micrograms/kg, i.p.) were administered after 30 min, while PGE2 (100 micrograms/kg, p.o.) was administered immediately afterwards. The distance traveled by charcoal in small intestine was measured after 15 and 30 min of charcoal administration, diarrhoea was observed every 30-min for six hour after serotonin administration and the volume of intestinal fluid was measured after 30 min of PGE2 administration. RESULTS: Arque-Ajeeb (0.07 ml and 0.14 ml/kg) significantly inhibited the frequency of defaecation and decreased the propulsion of charcoal meal through the gastrointestinal tract, reduced the wetness of faecal droppings in serotonin-induced diarrhoea and also reduced the PGE2-induced small intestine enteropooling. CONCLUSION: Arque-Ajeeb may have potential to reduce the diarrhoea in rats. Thus the drug may prove to be an alternate remedy in diarrhoea. [Abstract/Link to Full Text]

Pepato MT, Baviera AM, Vendramini RC, Brunetti IL
Evaluation of toxicity after one-months treatment with Bauhinia forficata decoction in streptozotocin-induced diabetic rats.
BMC Complement Altern Med. 2004 Jun 8;47.
BACKGROUND: Previous experiments have shown that a decoction of Bauhinia forficata leaves reduces the changes in carbohydrate and protein metabolism that occur in rats with streptozotocin-induced diabetes. In the present investigation, the serum activities of enzymes known to be reliable toxicity markers were monitored in normal and streptozotocin-diabetic rats to discover whether the use of B. forficata decoction has toxic effects on liver, muscle or pancreas tissue or on renal microcirculation. METHODS: An experimental group of normal and streptozotocin-diabetic rats received an aqueous decoction of fresh B. forficata leaves (150 g/L) by mouth for 33 days while a control group of normal and diabetic rats received water for the same length of time. The serum activity of the toxicity markers lactate dehydrogenase, creatine kinase, amylase, angiotensin-converting enzyme and bilirubin were assayed before receiving B. forficata decoction and on day 19 and 33 of treatment. RESULTS: The toxicity markers in normal and diabetic rats were not altered by the diabetes itself nor by treatment with decoction. Whether or not they received B. forficata decoction the normal rats showed a significant increase in serum amylase activity during the experimental period while there was a tendency for the diabetic rats, both treated and untreated with decoction, to have lower serum amylase activities than the normal rats. CONCLUSIONS: Administration of an aqueous decoction of B. forficata is a potential treatment for diabetes and does not produce toxic effects measurable with the enzyme markers used in our study. [Abstract/Link to Full Text]

Yount G, Solfvin J, Moore D, Schlitz M, Reading M, Aldape K, Qian Y
In vitro test of external Qigong.
BMC Complement Altern Med. 2004 Mar 15;45.
BACKGROUND: Practitioners of the alternative medical practice 'external Qigong' generally claim the ability to emit or direct "healing energy" to treat patients. We investigated the ability of experienced Qigong practitioners to enhance the healthy growth of cultured human cells in a series of studies, each following a rigorously designed protocol with randomization, blinding and controls for variability. METHODS: Qigong practitioners directed healing intentionality toward normal brain cell cultures in a basic science laboratory. Qigong treatments were delivered for 20 minutes from a minimum distance of 10 centimeters. Cell proliferation was measured by a standard colony-forming efficiency (CFE) assay and a CFE ratio (CFE for treated samples/CFE for sham samples) was the dependent measure for each experiment. RESULTS: During a pilot study (8 experiments), a trend of increased cell proliferation in Qigong-treated samples (CFE Qigong/sham ratios > 1.0) was observed (P = 0.162). In a formal study (28 experiments), a similar trend was observed, with Qigong-treated samples showing on average more colony formation than sham samples (P = 0.036). In a replication study (60 experiments), no significant difference between Qigong-treated samples and sham samples was observed (P = 0.465). CONCLUSION: We observed an apparent increase in the proliferation of cultured cells following external Qigong treatment by practitioners under strictly controlled conditions, but we did not observe this effect in a replication study. These results suggest the need for more controlled and thorough investigation of external Qigong before scientific validation is claimed. [Abstract/Link to Full Text]

Streitberger K, Witte S, Mansmann U, Knauer C, Kr�mer J, Scharf HP, Victor N
Efficacy and safety of acupuncture for chronic pain caused by gonarthrosis: a study protocol of an ongoing multi-centre randomised controlled clinical trial [ISRCTN27450856].
BMC Complement Altern Med. 2004 Mar 24;46.
BACKGROUND: Controlled clinical trials produced contradictory results with respect to a specific analgesic effect of acupuncture. There is a lack of large multi-centre acupuncture trials. The German Acupuncture Trial represents the largest multi-centre study of acupuncture in the treatment of chronic pain caused by gonarthrosis up to now. METHODS: 900 patients will be randomised to three treatment arms. One group receives verum acupuncture, the second sham acupuncture, and the third conservative standard therapy. The trial protocol is described with eligibility criteria, detailed information on the treatment definition, blinding, endpoints, safety evaluation, statistical methods, sample size determination, monitoring, legal aspects, and the current status of the trial. DISCUSSION: A critical discussion is given regarding the considerations about standardisation of the acupuncture treatment, the choice of the control group, and the blinding of patients and observers. [Abstract/Link to Full Text]

Baldwin CM, Kroesen K, Trochim WM, Bell IR
Complementary and conventional medicine: a concept map.
BMC Complement Altern Med. 2004 Feb 3;42.
BACKGROUND: Despite the substantive literature from survey research that has accumulated on complementary and alternative medicine (CAM) in the United States and elsewhere, very little research has been done to assess conceptual domains that CAM and conventional providers would emphasize in CAM survey studies. The objective of this study is to describe and interpret the results of concept mapping with conventional and CAM practitioners from a variety of backgrounds on the topic of CAM. METHODS: Concept mapping, including free sorts, ratings, and multidimensional scaling was used to organize conceptual domains relevant to CAM into a visual "cluster map." The panel consisted of CAM providers, conventional providers, and university faculty, and was convened to help formulate conceptual domains to guide the development of a CAM survey for use with United States military veterans. RESULTS: Eight conceptual clusters were identified: 1) Self-assessment, Self-care, and Quality of Life; 2) Health Status, Health Behaviors; 3) Self-assessment of Health; 4) Practical/Economic/ Environmental Concerns; 5) Needs Assessment; 6) CAM vs. Conventional Medicine; 7) Knowledge of CAM; and 8) Experience with CAM. The clusters suggest panelists saw interactions between CAM and conventional medicine as a critical component of the current medical landscape. CONCLUSIONS: Concept mapping provided insight into how CAM and conventional providers view the domain of health care, and was shown to be a useful tool in the formulation of CAM-related conceptual domains. [Abstract/Link to Full Text]

Singh V, Raidoo DM, Harries CS
The prevalence, patterns of usage and people's attitude towards complementary and alternative medicine (CAM) among the Indian community in Chatsworth, South Africa.
BMC Complement Altern Med. 2004 Feb 4;43.
BACKGROUND: The purpose of this study was to determine, among the Indian community of Chatsworth, South Africa, the prevalence and utilisation patterns of complementary and alternative medicine (CAM), attitudes associated with CAM use and communication patterns of CAM users with their primary care doctors. METHODS: Face-to-face structured interviews were conducted in Chatsworth, a suburb of Durban in which South Africans of Indian origin predominantly reside. Participants were 200 randomly selected adult English-speaking Indian residents. RESULTS: The prevalence of CAM usage for period 2000/2001 was 38.5% (95% confidence interval 31.7% to 45.6%). Spiritual healing and herbal/natural medicines, including vitamins were the most common types of CAM used, accounting for 42.8% and 48.1% respectively of overall CAM usage. People used CAM to treat conditions including diabetes mellitus, headaches, arthritis and joint pains, stress, skin disorders, backaches, hypertension and nasal disorders. Half of the CAM users used allopathic medicines concurrently. The cost of CAM utilization over this 1-year period, incurred by 80.5% of users for the duration of therapy for their most troublesome condition was below R500 (approximately US50 dollars). Age, sex, marital status, religion, level of education and income were shown not to influence the use of CAM. Greater than half (51.9%) of CAM users did so either upon the advice of someone they knew, or after noticing a CAM advertisement in the local press. Seventy-nine percent of CAM users indicated that they had positive outcomes with their treatments. Fifty four percent of CAM users (excluding those using spiritual healing only) failed to inform their doctors that they used CAM. The main reason given by half of this group was that informing their doctors did not seem necessary. CONCLUSION: The prevalence of CAM in Chatsworth is similar to findings in other parts of the world. Although CAM was used to treat many different ailments, this practice could not be attributed to any particular demographic profile. The majority of CAM users were satisfied with the effects of CAM. Findings support a need for greater integration of allopathic medicine and CAM, as well as improved communication between patients and caregivers regarding CAM usage. [Abstract/Link to Full Text]

Neese S, La Grange L, Trujillo E, Romero D
The effects of ethanol and silymarin treatment during gestation on spatial working memory.
BMC Complement Altern Med. 2004 Feb 12;44.
BACKGROUND: Using a rat model we have found that the bioflavonoid silymarin (SY) ameliorates some of the negative consequences of in utero exposure to ethanol (EtOH). In the current study our aim was to determine if spatial working memory (SWM) was impaired in offspring whose mothers were maintained on a liquid diet containing EtOH during different gestational weeks. We also determined if SWM was altered with a concomitant administration of SY with EtOH during specific gestational weeks. METHODS: We provided pregnant Fischer/344 rats with liquid diets containing 35% EtOH derived calories (EDC) during specific weeks of the gestational period. A silymarin/phospholipid compound containing 29.8% silybin co-administered with EtOH was also administered during specific weeks of the gestational period. We tested SWM of the offspring with a radial arm maze on postnatal day (PND) 60. After testing the rats were sacrificed and their brains perfused for later analysis. RESULTS: We observed SWM deficits, as well as a significantly lower brain weight in female offspring born of mothers treated with EtOH during the third week of gestation in comparison to mothers treated during either the first or second weeks of gestation. Rats from any group receiving EtOH in co-administration with SY showed no significant deficits in SWM. CONCLUSION: EtOH treatment during the last week of gestation had the greatest impact on SWM. The addition of SY to the EtOH liquid diet appeared to ameliorate the EtOH-induced learning deficits. [Abstract/Link to Full Text]

Recent Articles in Evidence-based Complementary and Alternative Medicine

Tohda C, Nakayama N, Hatanaka F, Komatsu K
Comparison of Anti-inflammatory Activities of Six Curcuma Rhizomes: A Possible Curcuminoid-independent Pathway Mediated by Curcuma phaeocaulis Extract.
Evid Based Complement Alternat Med. 2006 Jun;3(2):255-60.
We aimed to compare the anti-inflammatory activities of six species of Curcuma drugs using adjuvant arthritis model mice. When orally administered 1 day before the injection of adjuvant, the methanol extract of Curcuma phaeocaulis significantly inhibited paw swelling and the serum haptoglobin concentration in adjuvant arthritis mice. Also when orally administered 1 day after the injection of adjuvant, the methanol extract of Curcuma phaeocaulis significantly inhibited paw swelling. Other Curcuma species (Curcuma longa, Curcuma wenyujin, Curcuma kwangsiensis, Curcuma zedoaria and Curcuma aromatica) had no significant inhibitory effects on adjuvant-induced paw swelling. Cyclooxygenase (COX)-2 activity was significantly inhibited by the methanol extract of C. phaeocaulis. Curcuminoids' (curcumin, bis-demethoxycurcumin and demethoxycurcumin) were rich in C. longa, but less in C. phaeocaulis and C. aromatica, not in C. wenyujin, C. kwangsiensis and C. zedoaria, suggesting that curcuminoids' contents do not relate to inhibition of arthritis swelling. Therefore, C. phaeocaulis may be a useful drug among Curcuma species for acute inflammation, and the active constituents of C. phaeocaulis are not curcuminoids. [Abstract/Link to Full Text]

Trusheva B, Popova M, Bankova V, Simova S, Marcucci MC, Miorin PL, da Rocha Pasin F, Tsvetkova I
Bioactive constituents of brazilian red propolis.
Evid Based Complement Alternat Med. 2006 Jun;3(2):249-54.
In a new propolis type, red Brazilian propolis, 14 compounds were identified (six of them new for propolis), among them simple phenolics, triterepenoids, isoflavonoids, prenylated benzophenones and a naphthoquinone epoxide (isolated for the first time from a natural source). Three of the major components demonstrated significant antimicrobial activity, and two (obtained as inseparable mixture) possessed radical scavenging activity against 1,1-diphenyl-2-picrylhydrazyl (DPPH). [Abstract/Link to Full Text]

Ravindranath MH, Saravanan TS, Monteclaro CC, Presser N, Ye X, Selvan SR, Brosman S
Epicatechins Purified from Green Tea (Camellia sinensis) Differentially Suppress Growth of Gender-Dependent Human Cancer Cell Lines.
Evid Based Complement Alternat Med. 2006 Jun;3(2):237-47.
The anticancer potential of catechins derived from green tea is not well understood, in part because catechin-related growth suppression and/or apoptosis appears to vary with the type and stage of malignancy as well as with the type of catechin. This in vitro study examined the biological effects of epicatechin (EC), epigallocatechin (EGC), EC 3-gallate (ECG) and EGC 3-gallate (EGCG) in cell lines from human gender-specific cancers. Cell lines developed from organ-confined (HH870) and metastatic (DU145) prostate cancer, and from moderately (HH450) and poorly differentiated (HH639) epithelial ovarian cancer were grown with or without EC, EGC, ECG or EGCG. When untreated cells reached confluency, viability and doubling time were measured for treated and untreated cells. Whereas EC treatment reduced proliferation of HH639 cells by 50%, EGCG suppressed proliferation of all cell lines by 50%. ECG was even more potent: it inhibited DU145, HH870, HH450 and HH639 cells at concentrations of 24, 27, 29 and 30 microM, whereas EGCG inhibited DU145, HH870, HH450 and HH639 cells at concentrations 89, 45, 62 and 42 microM. When compared with EGCG, ECG more effectively suppresses the growth of prostate cancer and epithelial ovarian cancer cell lines derived from tumors of patients with different stages of disease. [Abstract/Link to Full Text]

Azaizeh H, Saad B, Khalil K, Said O
The state of the art of traditional arab herbal medicine in the eastern region of the mediterranean: a review.
Evid Based Complement Alternat Med. 2006 Jun;3(2):229-35.
Historical and current studies indicate that the Eastern region of the Mediterranean has been distinguished from other regions by a rich inventory of complementary alternative medicine (CAM), in particular herbal medicine. Data collected from several surveys and studies indicate that there is a flourishing and well-developed trade of herbs. These surveys also reveal that 200-250 herbs are used in treating human diseases and are sold or traded in market places in the Mediterranean region or internationally. In addition, some of these herbs are rare or even endangered species. In regard to the status of the know-how of herbalists, unfortunately, herbal medicine in our region is mostly prescribed by ethnopharmacologists symptomatically-based on signs and symptoms alone, rather than as a result of a full understanding of the underlying disease. In some cases, herbs used today may not even correspond to the plants described originally in the old literature, as the former are cultivated from herbs that went through different breeding procedures throughout several centuries. This article presents a systematic review of both the state of the art of traditional Arab herbal medicine and the status of the know-how of Arab herbalists. [Abstract/Link to Full Text]

Nigam Y, Bexfield A, Thomas S, Ratcliffe NA
Maggot Therapy: The Science and Implication for CAM Part I-History and Bacterial Resistance.
Evid Based Complement Alternat Med. 2006 Jun;3(2):223-7.
It is now a universally acknowledged fact that maggot therapy can be used successfully to treat chronic, long-standing, infected wounds, which have previously failed to respond to conventional treatment. Such wounds are typically characterized by the presence of necrotic tissue, underlying infection and poor healing. Maggot therapy employs the use of freshly emerged, sterile larvae of the common green-bottle fly, Phaenicia (Lucilia) sericata, and is a form of artificially induced myiasis in a controlled clinical situation. In this review article, we will discuss the role of maggots and their preparation for clinical use. Maggot therapy has the following three core beneficial effects on a wound: debridement, disinfection and enhanced healing. In part I we explore our current understanding of the mechanisms underlying these effects. [Abstract/Link to Full Text]

Rastogi S, Kulshreshtha DK, Rawat AK
Streblus asper Lour. (Shakhotaka): A Review of its Chemical, Pharmacological and Ethnomedicinal Properties.
Evid Based Complement Alternat Med. 2006 Jun;3(2):217-22.
Streblus asper Lour is a small tree found in tropical countries, such as India, Sri Lanka, Malaysia, the Philippines and Thailand. Various parts of this plant are used in Ayurveda and other folk medicines for the treatment of different ailments such as filariasis, leprosy, toothache, diarrhea, dysentery and cancer. Research carried out using different in vitro and in vivo techniques of biological evaluation support most of these claims. This review presents the botany, chemistry, traditional uses and pharmacology of this medicinal plant. [Abstract/Link to Full Text]

Vojdani A, Erde J
Regulatory T Cells, a Potent Immunoregulatory Target for CAM Researchers: Modulating Allergic and Infectious Disease Pathology (II).
Evid Based Complement Alternat Med. 2006 Jun;3(2):209-15.
Regulatory T (T(reg)) cells maintain dominant control of immune responses to foreign materials and microbes. Appropriate T(reg) cell suppression of immune responses is essential for the maintenance of efficacious defensive responses and the limitation of collateral tissue damage due to excess inflammation. Allergy and infection are well studied and frequent afflictions in which T(reg) cells play an essential role. As such, they provide excellent models to communicate the significance and relevance of T(reg) cells to complementary and alternative medicine (CAM). [Abstract/Link to Full Text]

Usichenko TI, Edinger H, Gizhko VV, Lehmann C, Wendt M, Feyerherd F
Low-intensity electromagnetic millimeter waves for pain therapy.
Evid Based Complement Alternat Med. 2006 Jun;3(2):201-7.
Millimeter wave therapy (MWT), a non-invasive complementary therapeutic technique is claimed to possess analgesic properties. We reviewed the clinical studies describing the pain-relief effect of MWT. Medline-based search according to review criteria and evaluation of methodological quality of the retrieved studies was performed. Of 13 studies, 9 of them were randomized controlled trials (RCTs), only three studies yielded more than 3 points on the Oxford scale of methodological quality of RCTs. MWT was reported to be effective in the treatment of headache, arthritic, neuropathic and acute postoperative pain. The rapid onset of pain relief during MWT lasting hours to days after, remote to the site of exposure (acupuncture points), was the most characteristic feature in MWT application for pain relief. The most commonly used parameters of MWT were the MW frequencies between 30 and 70 GHz and power density up to 10 mW cm(-2). The promising results from pilot case series studies and small-size RCTs for analgesic/hypoalgesic effects of MWT should be verified in large-scale RCTs on the effectiveness of this treatment method. [Abstract/Link to Full Text]

Ohnishi ST, Ohnishi T
The Nishino Breathing Method and Ki-energy (Life-energy): A Challenge to Traditional Scientific Thinking.
Evid Based Complement Alternat Med. 2006 Jun;3(2):191-200.
The breathing method, which was developed and is being taught by Kozo Nishino, a Japanese Ki-expert, is for raising the levels of Ki-energy (life-energy or the vitality) of an individual. It is neither a therapy nor a healing technique. However, many of his students have experienced an improvement in their health, and in some cases, they were able to overcome health problems by themselves. Since this is an interesting subject from the standpoint of complementary and alternative medicine (CAM), we have been collaborating with Nishino to conduct a scientific investigation of his Ki-energy. We found that Nishino's Ki-energy can inhibit cell division of cancer cells, protect isolated mitochondria from heat deterioration and reduce lipid peroxidation in heat-treated mitochondria. Although Ki-energy may consist of several different energy forms, we found that at least one of them is near-infrared radiation between the wavelength range of 0.8 and 2.7 microm. Another interesting observation at his school is the Taiki-practice (paired Ki-practice). During this practice, Nishino can 'move' his students without any physical contact. Many of them run, jump or roll on the floor when they receive his Ki-energy. We studied this and propose that 'information' is conveyed through the air between two individuals by Ki-energy. This may be called a five sense-independent, life-to-life communication by Ki. All of our results suggest that we should re-evaluate the Cartesian dualism (separation of mind and body) which has been a fundamental principle of modern science for the past three centuries. [Abstract/Link to Full Text]

Bennett MP, Lengacher C
Humor and Laughter May Influence Health: II. Complementary Therapies and Humor in a Clinical Population.
Evid Based Complement Alternat Med. 2006 Jun;3(2):187-90.
Our results support a connection between sense of humor and self-reported physical health, however, it is difficult to determine the relationship to any specific disease process. Whereas relationships between sense of humor and self-reported measures of physical well-being appear to be supported, more research is required to determine interrelationships between sense of humor and well-being. [Abstract/Link to Full Text]

Bellavite P, Ortolani R, Conforti A
Immunology and homeopathy. 3. Experimental studies on animal models.
Evid Based Complement Alternat Med. 2006 Jun;3(2):171-86.
A search of the literature and the experiments carried out by the authors of this review show that there are a number of animal models where the effect of homeopathic dilutions or the principles of homeopathic medicine have been tested. The results relate to the immunostimulation by ultralow doses of antigens, the immunological models of the 'simile', the regulation of acute or chronic inflammatory processes and the use of homeopathic medicines in farming. The models utilized by different research groups are extremely etherogeneous and differ as the test medicines, the dilutions and the outcomes are concerned. Some experimental lines, particularly those utilizing mice models of immunomodulation and anti-inflammatory effects of homeopathic complex formulations, give support to a real effect of homeopathic high dilutions in animals, but often these data are of preliminary nature and have not been independently replicated. The evidence emerging from animal models is supporting the traditional 'simile' rule, according to which ultralow doses of compounds, that in high doses are pathogenic, may have paradoxically a protective or curative effect. Despite a few encouraging observational studies, the effectiveness of the homeopathic prevention or therapy of infections in veterinary medicine is not sufficiently supported by randomized and controlled trials. [Abstract/Link to Full Text]

Cooper EL
Stem Cells and CAM.
Evid Based Complement Alternat Med. 2006 Jun;3(2):167-9. [Abstract/Link to Full Text]

Lewith GT, Verhoef MJ
The International Society for Complementary Medicine Research (ISCMR): the way forward.
Evid Based Complement Alternat Med. 2006 Mar;3(1):157-8.
This article describes the birth and development of the International Society for Complementary Medicine Research (ISCMR) from its inception in 2003. The society's main function is to facilitate the development of CAM research internationally and to use its networks, website, news flashes and newsletters to communicate with its members and bring CAM researchers together from all over the world. [Abstract/Link to Full Text]

Kaphle K, Wu LS, Yang NY, Lin JH
Herbal medicine research in Taiwan.
Evid Based Complement Alternat Med. 2006 Mar;3(1):149-55.
Of all the countries in the world, why did you choose Taiwan to pursue your study? It is a question that I (comments of the first author) have answered a thousand times. My first visit to a laboratory at National Taiwan University opened my eyes to the possibilities of herbal medicine research, especially in the area of veterinary medicine. It became my ambition to link the knowledge of Traditional Chinese Medicine (TCM) and Ayurveda from the Indian subcontinent and their integration with other systems of medicine, including Western medicine (WM), to achieve the concept of Sustainable Medicine, firstly for animals and then for humans. The Ministry of Economic Affairs (MOEA) has implemented a technology development program to quickly establish the key technologies, and this is a moment of opportunity for Taiwan's traditional herbal medicine industry to upgrade and transform itself. This paper, initially intended to be a student's narration, has evolved into a multi-author treatise on the present state and likely future scenario of herbal medicine research in Taiwan. [Abstract/Link to Full Text]

Fisher P
Homeopathy and The Lancet.
Evid Based Complement Alternat Med. 2006 Mar;3(1):145-7. [Abstract/Link to Full Text]

Tsao JC
CAM for pediatric pain: what is state-of-the-research?
Evid Based Complement Alternat Med. 2006 Mar;3(1):143-4.
Previously, we reviewed the evidence for the efficacy of CAM approaches for pediatric pain (volume 2; issue 2; 2005) using criteria developed by the American Psychological Association Division 12 Task Force. Our review focused on CAM modalities that had been tested with at least one controlled trial or multiple baseline study. In addition, only those trials in which children comprised the study sample were included. Thus, several CAM modalities were not included in our review. Key ethical and other reasons for the limited literature on CAM for pediatric pain as well as directions for future studies are discussed. [Abstract/Link to Full Text]

Hankey A
CAM and the phenomenology of pain.
Evid Based Complement Alternat Med. 2006 Mar;3(1):139-41.
Many CAM modalities afford relief from pain, each in its own way, or according to its own terminology. Comparison of different CAM modalities results in a simple phenomenology of pain centered around the idea that pain may be associated with blockages of the flow of energy in the system of nadis/acupuncture meridians. [Abstract/Link to Full Text]

Adams JD, Garcia C
Women's health among the Chumash.
Evid Based Complement Alternat Med. 2006 Mar;3(1):125-31.
Plants were, and still are, widely used for a number of conditions affecting women in California. This article discusses traditional remedies of the Chumash for dysmenorrhea, premenstrual syndrome, feminine hygiene, heavy menstruation, urinary tract infections, parturition, lactation, infant care, menopause, sexually transmitted diseases, fertility, contraception and abortions. Many plants are presented including Artemisia douglasiana, Paeonia californica, Trichostema lanatum, Salvia apiana, Ephedra viridis, Leymus condensatus, Vitis californica, Eschscholzia californica, Rosa californica, Scirpus acutus, Anemopsis californica and Phoradendron macrophyllum. By providing the specific uses of plants for specific diseases and discussing chemistry, efficacy and safety concerns for each plant, we hope that this article gives direction to women seeking to use plants in their health care. [Abstract/Link to Full Text]

Adams LS, Seeram NP, Hardy ML, Carpenter C, Heber D
Analysis of the interactions of botanical extract combinations against the viability of prostate cancer cell lines.
Evid Based Complement Alternat Med. 2006 Mar;3(1):117-24.
Herbal medicines are often combinations of botanical extracts that are assumed to have additive or synergistic effects. The purpose of this investigation was to compare the effect of individual botanical extracts with combinations of extracts on prostate cell viability. We then modeled the interactions between botanical extracts in combination isobolographically. Scutellaria baicalensis, Rabdosia rubescens, Panax-pseudo ginseng, Dendranthema morifolium, Glycyrrhiza uralensis and Serenoa repens were collected, taxonomically identified and extracts prepared. Effects of the extracts on cell viability were quantitated in prostate cell lines using a luminescent ATP cell viability assay. Combinations of two botanical extracts of the four most active extracts were tested in the 22Rv1 cell line and their interactions assessed using isobolographic analysis. Each extract significantly inhibited the proliferation of prostate cell lines in a time- and dose-dependent manner except S. repens. The most active extracts, S. baicalensis, D. morifolium, G. uralensis and R. rubescens were tested as two-extract combinations. S. baicalensis and D. morifolium when combined were additive with a trend toward synergy, whereas D. morifolium and R. rubescens together were additive. The remaining two-extract combinations showed antagonism. The four extracts together were significantly more effective than the two-by-two combinations and the individual extracts alone. Combining the four herbal extracts significantly enhanced their activity in the cell lines tested compared with extracts alone. The less predictable nature of the two-way combinations suggests a need for careful characterization of the effects of each individual herb based on their intended use. [Abstract/Link to Full Text]

Takahashi T, Yu F, Zhu SJ, Moriya J, Sumino H, Morimoto S, Yamaguchi N, Kanda T
Beneficial effect of brewers' yeast extract on daily activity in a murine model of chronic fatigue syndrome.
Evid Based Complement Alternat Med. 2006 Mar;3(1):109-15.
The aim of this study was to assess the effect of Brewers' yeast extract (BYE) on daily activity in a mouse model of chronic fatigue syndrome (CFS). CFS was induced by repeated injection of Brucella abortus (BA) antigen every 2 weeks. BYE was orally administered to mice in a dose of 2 g per kg per day for 2 weeks before injecting BA and for 4 weeks thereafter. We evaluated daily running activity in mice receiving BYE as compared with that in untreated mice. Weekly variation of body weight (BW) and survival in both groups was monitored during the observation period. Spleen weight (SW), SW/BW ratio, percent splenic follicular area and expression levels of interferon-gamma (IFN-gamma) and interleukin-10 (IL-10) mRNA in spleen were determined in both groups at the time of sacrifice. The daily activity during 2 weeks after the second BA injection was significantly higher in the treated group than in the control. There was no difference in BW between both groups through the experimental course. Two mice in the control died 2 and 7 days after the second injection, whereas no mice in the treated group died. Significantly decreased SW and SW/BW ratio were observed in the treated mice together with elevation of splenic follicular area. There were suppressed IFN-gamma and IL-10 mRNA levels in spleens from the treated mice. Our results suggest that BYE might have a protective effect on the marked reduction in activity following repeated BA injection via normalization of host immune responses. [Abstract/Link to Full Text]

Belon P, Banerjee P, Choudhury SC, Banerjee A, Biswas SJ, Karmakar SR, Pathak S, Guha B, Chatterjee S, Bhattacharjee N, Das JK, Khuda-Bukhsh AR
Can administration of potentized homeopathic remedy, Arsenicum album, alter antinuclear antibody (ANA) titer in people living in high-risk arsenic contaminated areas? I. A correlation with certain hematological parameters.
Evid Based Complement Alternat Med. 2006 Mar;3(1):99-107.
To examine whether elevated antinuclear antibody (ANA) titers reported in random human population of arsenic contaminated villages can be reverted to the normal range by administration of a potentized homeopathic drug, Arsenicum album, randomly selected volunteers in two arsenic contaminated villages and one arsenic-free village in West Bengal (India) were periodically tested for their ANA titer as well as various blood parameters in two types of experiments: 'placebo-controlled double blind' experiment for shorter duration and 'uncontrolled verum fed experiment' for longer duration. Positive modulation of ANA titer was observed along with changes in certain relevant hematological parameters, namely total count of red blood cells and white blood cells, packed cell volume, hemoglobin content, erythrocyte sedimentation rate and blood sugar level, mostly within 2 months of drug administration. Thus, Arsenicum album appears to have great potential for ameliorating arsenic induced elevated ANA titer and other hematological toxicities. [Abstract/Link to Full Text]

Saad B, Dakwar S, Said O, Abu-Hijleh G, Al Battah F, Kmeel A, Aziazeh H
Evaluation of medicinal plant hepatotoxicity in co-cultures of hepatocytes and monocytes.
Evid Based Complement Alternat Med. 2006 Mar;3(1):93-8.
Non-parenchymal cells might play an important role in the modulation of xenobiotic metabolism in liver and its pharmacological and toxicological consequences. Therefore, the role of cell-to-cell interactions in herbal induced liver toxicity was investigated in monocultures of cells from the human hepatocyte cell line (HepG2) and in co-cultures of cells from the HepG2 cell line and cells from the human monocyte cell line (THP1). Cells were treated with various concentrations (1-500 microg ml(-1)) of extracts of Pistacia palaestina, Juglans regia and Quercus ithaburensis for 24 h. Extracts from Cleome droserifolia, a known toxic plant, were taken as positive control. In the co-culture system, toxic effects were observed after exposure to extracts of Pistacia palestina and C. droserifolia. These two extracts significantly reduced by cell viability as measured the MTT test and the LDH assay. Whereas in hepatocyte cultures, only extracts of C. droserifolia were found to affect the cell viability. The production levels of albumin from hepatocytes were not affected by treatment with plant extracts in both culture systems. It seems that the observed reduction in cell viability after exposure to extracts of P. palestina in co-cultures but not in monocultures is a result of monocyte-derived factors. The use of liver cell co-cultures is therefore a useful approach to investigate the influence of intercellular communication on xenobiotic metabolism in liver. [Abstract/Link to Full Text]

Al-Fatimi MA, J�lich WD, Jansen R, Lindequist U
Bioactive components of the traditionally used mushroom Podaxis pistillaris.
Evid Based Complement Alternat Med. 2006 Mar;3(1):87-92.
In the course of an ethnobotanical study on fungi used in Yemeni ethnomedicine the fungus Podaxis pistillaris (Podaxales, Podaxaceae, Basidiomycetes) was found to exhibit antibacterial activity against Staphylococcus aureus, Micrococcus flavus, Bacillus subtilis, Proteus mirabilis, Serratia marcescens and Escherichia coli. In the culture medium of P. pistillaris three epidithiodiketopiperazines were identified by activity-guided isolation. Based on spectral data (NMR, ESI-MS and DCI-MS) their identity was established as epicorazine A (1), epicorazine B (2) and epicorazine C (3, antibiotic F 3822), which have not been reported as constituents of P. pistillaris previously. It is assumed that the identified compounds contribute to the antibacterial activity of the extract. [Abstract/Link to Full Text]

Joshi H, Parle M
Brahmi rasayana improves learning and memory in mice.
Evid Based Complement Alternat Med. 2006 Mar;3(1):79-85.
Cure of cognitive disorders such as amnesia, attention deficit and Alzheimer's disease is still a nightmare in the field of medicine. Nootropic agents such as piracetam, aniracetam and choline esterase inhibitors like Donepezil are being used to improve memory, mood and behavior, but the resulting side effects associated with these agents have made their use limited. The present study was undertaken to assess the potential of Brahmi rasayana (BR) as a memory enhancer. BR (100 and 200 mg kg(-1) p.o.) was administered for eight successive days to both young and aged mice. Elevated plus maze and passive-avoidance paradigm were employed to evaluate learning and memory parameters. Scopolamine (0.4 mg kg(-1) i.p.) was used to induce amnesia in mice. The effect of BR on whole brain AChE activity was also assessed. Piracetam (200 mg kg(-1) i.p.) was used as a standard nootropic agent. BR significantly improved learning and memory in young mice and reversed the amnesia induced by both scopolamine (0.4 mg kg(-1) i.p.) and natural aging. BR significantly decreased whole brain acetyl cholinesterase activity. BR might prove to be a useful memory restorative agent in the treatment of dementia seen in elderly. [Abstract/Link to Full Text]

Inokuchi Y, Shimazawa M, Nakajima Y, Suemori S, Mishima S, Hara H
Brazilian green propolis protects against retinal damage in vitro and in vivo.
Evid Based Complement Alternat Med. 2006 Mar;3(1):71-7.
Propolis, a honeybee product, has gained popularity as a food and alternative medicine. Its constituents have been shown to exert pharmacological (anticancer, antimicrobial and anti-inflammatory) effects. We investigated whether Brazilian green propolis exerts neuroprotective effects in the retina in vitro and/or in vivo. In vitro, retinal damage was induced by 24 h hydrogen peroxide (H2O2) exposure, and cell viability was measured by Hoechst 33342 and YO-PRO-1 staining or by a resazurin-reduction assay. Propolis inhibited the neurotoxicity and apoptosis induced in cultured retinal ganglion cells (RGC-5, a rat ganglion cell line transformed using E1A virus) by 24 h H2O2 exposure. Propolis also inhibited the neurotoxicity induced in RGC-5 cultures by staurosporine. Regarding the possible underlying mechanism, in pig retina homogenates propolis protected against oxidative stress (lipid peroxidation), as also did trolox (water-soluble vitamin E). In mice in vivo, propolis (100 mg kg(-1); intraperitoneally administered four times) reduced the retinal damage (decrease in retinal ganglion cells and in thickness of inner plexiform layer) induced by intravitreal in vivo N-methyl-d-aspartate injection. These findings indicate that Brazilian green propolis has neuroprotective effects against retinal damage both in vitro and in vivo, and that a propolis-induced inhibition of oxidative stress may be partly responsible for these neuroprotective effects. [Abstract/Link to Full Text]

Katz S, Harris R, Lau JT, Chau A
The use of gene expression analysis and proteomic databases in the development of a screening system to determine the value of natural medicinal products.
Evid Based Complement Alternat Med. 2006 Mar;3(1):65-70.
A rapid throughput screening system involving gene expression analysis was developed in order to investigate the potential of bioactive chemicals contained in natural health products as effective drug therapy, in particular the ability of these chemicals to alleviate the inflammatory response in human airway epithelial cells. A number of databases were searched to retrieve the information needed to properly analyze the gene expression profiles obtained. The gene expression of human bronchial epithelial cells infected with rhinovirus and/or exposed to platelet activating factor was analyzed. Following analysis of the gene expression data the total number of expressed proteins that may potentially act as a marker for monitoring the modulation of airway inflammation was narrowed to 19. Further studies will involve selecting antibodies for these proteins, culturing airway epithelial cells in the presence of extracts of natural health products, extracting the proteins and identifying them by western blot analysis. [Abstract/Link to Full Text]

Bennett MP, Lengacher CA
Humor and laughter may influence health. I. History and background.
Evid Based Complement Alternat Med. 2006 Mar;3(1):61-3.
Articles in both the lay and professional literature have extolled the virtues of humor, many giving the impression that the health benefits of humor are well documented by the scientific and medical community. The concept that humor or laughter can be therapeutic goes back to biblical times and this belief has received varying levels of support from the scientific community at different points in its history. Current research indicates that using humor is well accepted by the public and is frequently used as a coping mechanism. However, the scientific evidence of the benefits of using humor on various health related outcomes still leaves many questions unanswered. [Abstract/Link to Full Text]

Lonsdale D
A review of the biochemistry, metabolism and clinical benefits of thiamin(e) and its derivatives.
Evid Based Complement Alternat Med. 2006 Mar;3(1):49-59.
Thiamin(e), also known as vitamin B1, is now known to play a fundamental role in energy metabolism. Its discovery followed from the original early research on the 'anti-beriberi factor' found in rice polishings. After its synthesis in 1936, it led to many years of research to find its action in treating beriberi, a lethal scourge known for thousands of years, particularly in cultures dependent on rice as a staple. This paper refers to the previously described symptomatology of beriberi, emphasizing that it differs from that in pure, experimentally induced thiamine deficiency in human subjects. Emphasis is placed on some of the more unusual manifestations of thiamine deficiency and its potential role in modern nutrition. Its biochemistry and pathophysiology are discussed and some of the less common conditions associated with thiamine deficiency are reviewed. An understanding of the role of thiamine in modern nutrition is crucial in the rapidly advancing knowledge applicable to Complementary Alternative Medicine. References are given that provide insight into the use of this vitamin in clinical conditions that are not usually associated with nutritional deficiency. The role of allithiamine and its synthetic derivatives is discussed. Thiamine plays a vital role in metabolism of glucose. Thus, emphasis is placed on the fact that ingestion of excessive simple carbohydrates automatically increases the need for this vitamin. This is referred to as high calorie malnutrition. [Abstract/Link to Full Text]

Elisabetsky E, Costa-Campos L
The alkaloid alstonine: a review of its pharmacological properties.
Evid Based Complement Alternat Med. 2006 Mar;3(1):39-48.
Indole compounds, related to the metabolism of tryptophan, constitute an extensive family, and are found in bacteria, plants and animals. Indolic compounds possess significant and complex physiological roles, and especially indole alkaloids have historically constituted a class of major importance in the development of new plant derived drugs. The indole alkaloid alstonine has been identified as the major component of a plant-based remedy, used in Nigeria to treat mental illnesses by traditional psychiatrists. Although it is certainly difficult to compare the very concept of mental disorders in different cultures, the traditional use of alstonine is remarkably compatible with its profile in experimental animals. Even though alstonine in mice models shows a psychopharmacological profile closer to the newer atypical antipsychotic agents, it also shows important differences and what seems to be an exclusive mechanism of action, not entirely clarified at this point. Considering the seemingly unique mode of action of alstonine and that its traditional use can be viewed as indicative of bioavailability and safety, this review focuses on the effects of alstonine in the central nervous system, particularly on its unique profile as an antipsychotic agent. We suggest that a thorough understanding of traditional medical concepts of health and disease in general and traditional medical practices in particular, can lead to true innovation in paradigms of drug action and development. Overall, the study of this unique indole alkaloid may be considered as another example of the richness of medicinal plants and traditional medical systems in the discovery of new prototypic drugs. [Abstract/Link to Full Text]

Seya T, Akazawa T, Tsujita T, Matsumoto M
Role of Toll-like receptors in adjuvant-augmented immune therapies.
Evid Based Complement Alternat Med. 2006 Mar;3(1):31-8; discussion 133-7.
Effective therapeutic vaccines contain two primary constituents, antigen and adjuvant. Adjuvants consisting of microbial pattern molecules play a central role in vaccination. Successful vaccine requires efficient induction of antibody (Ab), type I interferons (IFN), cytokines/chemokines, cytotoxic T lymphocytes (CTL) and/or NK cells. Toll-like receptors (TLRs) in myeloid dendritic cells (mDC) essentially act as adjuvant receptors and sustain the molecular basis of adjuvant activity. Current consensus is that TLRs and their adapters introduce signals to preferentially induce IFN-alpha/beta, chemokines and proinflammatory cytokines, and mature mDC to augment antigen presentation. Although most of these data were obtained with mice, the results are presumed to be adaptable to humans. Whenever TLR pathway is activated in mDC, NK and/or CTL activation is promoted. For induction of antigen-specific CTL toward phagocytosed material, cross-priming must be induced in mDC, which is also sustained by TLR signaling in mDC. Since the TLR responses vary with different adjuvants, mDC functions are skewed depending on adjuvant-specific direction of mDC maturation. It appears that the directed maturation of mDC largely relies on selection of appropriate sets of TLRs and their adapter signaling pathways. Synthetic chimera molecules consisting of TLR agonists and target antigens are found to be effective in induction of CTL to eliminate target cells in vivo. Here, we review the role of human TLRs and adapters in a variety of host immune responses. We will also describe the relevance of adjuvants in the manipulation of receptors and adapters in vaccine therapy. [Abstract/Link to Full Text]

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Recent Articles in Journal of Pharmacy & Pharmaceutical Sciences

Xiao Z, Hansen CB, Allen TM, Miller GG, Moore RB
Distribution of photosensitizers in bladder cancer spheroids: implications for intravesical instillation of photosensitizers for photodynamic therapy of bladder cancer.
J Pharm Pharm Sci. 2005;8(3):536-43.
PURPOSE: Uniform intratumor distribution of sufficient photosensitizer is one of the important aspects of photodynamic therapy for solid tumors. METHODS: Multicellular spheroids derived from a human transitional cell carcinoma cell line (MGHU3) were used as a surrogate system of tiny solid tumors to study intratumor distribution of photosensitizers. Photosensitizers included Photofrin, hypocrellins (HBEA-R1/R2, HBBA-R2), aluminum phthalocyanine chloride (AlPC), benzoporphyrin derivative monoacid ring A (BPD-MA), protoporphyrin-IX (PpIX), and liposomal formulations of HBBA-R2 and BPD-MA. Spheroids were incubated with various doses of the above drugs for 1-4 hours, and were examined by confocal microscopy. RESULTS: Histology showed all cells were healthy in spheroids less than 400 microm in diameter. Scanning electron microscopy showed tight cell-to-cell interdigitation in spheroids. HBEA-R1/R2 distributed more uniformly in spheroids than other drugs. Free hypocrellins and BPD-MA penetrated spheroids centripetally deeper than AlPC, Photofrin, and PpIX. Liposomal HBBA-R2 and BPD-MA penetrated less than their free formulations. CONCLUSIONS: The spheroids mimic solid tumors prior to neovascularization. Based on drug distribution in spheroids, hypocrellins and BPD-MA appear superior to Photofrin, AlPC and PpIX for intravesical administration for bladder cancer phototherapy. [Abstract/Link to Full Text]

Yu FR, Lian XZ, Guo HY, McGuire PM, Li RD, Wang R, Yu FH
Isolation and characterization of methyl esters and derivatives from Euphorbia kansui (Euphorbiaceae) and their inhibitory effects on the human SGC-7901 cells.
J Pharm Pharm Sci. 2005;8(3):528-35.
PURPOSE: In this study, the inhibitory activity of the methyl esters and derivatives extracted from Euphorbia kansui (Euphorbiaceae) and their effect on apoptosis and cell cycle distribution in the human gastric cancer cell line (SGC-7901) were evaluated. METHODS: The inhibitory activity of the methyl esters and derivatives was evaluated by using trypan-blue, MTT (3-(4, 5-dimethyl thiazol-2yl) - 2, 5-diphenyltetrazolium bromide), and FCM (flow cytometry) assays. 5-fluorouracile (5-FU) was used for a positive control. RESULTS: Six new methyl esters and derivatives were extracted from the root of E. kansui. Subjecting the SGC-7901 cell line to the extract indicated that methyl ester derivatives could initiate growth inhibition and induce apoptosis in these tumor cells. The inhibitory rates as measured from trypan-blue and MTT assays were significantly increased and are comparable to those of the common antitumor agent 5-FU. In addition, the methyl ester extract effectively inhibited the proliferation of SGC-7901 cells by interfering with the progression of the cells through the G1 phase of the cell cycle. CONCLUSION: The current study indicates that methyl esters might be a promising chemopreventive and chemotherapeutic agent for treating various forms of cancer by causing apoptosis and proliferation inhibition. [Abstract/Link to Full Text]

Jekerle V, Kassack MU, Reilly RM, Wiese M, Piquette-Miller M
Functional comparison of single- and double-stranded mdr1 antisense oligodeoxynucleotides in human ovarian cancer cell lines.
J Pharm Pharm Sci. 2005;8(3):516-27.
PURPOSE: P-glycoprotein mediated multidrug resistance presents a major obstacle in the successful therapeutic treatment of solid tumors such as ovarian cancer. Among the more promising techniques used to overcome multidrug resistance in ovarian cancer, is the transcriptional suppression of P-glycoprotein by antisense oligodeoxynucleotides (ODNs). To design more potent antisense ODNs, we explored the concept that double-stranded antisense ODNs may offer advantages in stability and potency over single-stranded in analogy to double-stranded siRNA. METHOD: Single-stranded phosphorothioate antisense ODNs against the human mdr1 gene were compared to the duplex of the active antisense and sense sequence of the same length. Concentration dependant effects on P-glycoprotein (Pgp) expression and functionality were quantitatively compared in the Pgp overexpressing ovarian cancer cell line A2780/Adr and its parental cell line A2780. Antisense ODNs were (111)Indium- and fluorescein isothiocyanate-conjugated for stability, cellular uptake and nuclear localization studies. Duplex formation significantly enhanced transcriptional inhibition of Pgp surface expression and functionality. Cellular uptake and distribution to the nucleus was improved when utilized as double-stranded DNA. CONCLUSION: Novel findings from this study suggest that double-stranded antisense ODNs more effectively inhibit target protein expression and consequently enhance chemoresponsiveness through improvements in cellular uptake and distribution to the nucleus. [Abstract/Link to Full Text]

Megalli S, Aktan F, Davies NM, Roufogalis BD
Phytopreventative anti-hyperlipidemic effects of gynostemma pentaphyllum in rats.
J Pharm Pharm Sci. 2005;8(3):507-15.
PURPOSE: Gynostemma pentaphyllum is widely used in traditional Chinese medicine. Preliminary studies indicate Gynostemma isolated triterpine glycosides lower cholesterol. Our studies examine anti-hyperlipidemic effects of gypenosides. METHODS: Gynostemma activity was examined in poloxamer P407 induced hyperlipidemia in rats. RESULTS: 1 g/kg P407 induced plasma triglyceride (25 fold), total cholesterol (6 fold), low density lipoprotein cholesterol (LDL) (7 fold), high density lipoprotein cholesterol (HDL) (1.6 fold), and nitrite (8 fold). After acute (4 days) and chronic (12 days) oral administration the gypenoside extract (250 mg/kg) reduced triglyceride (53% and 85%, respectively) and total cholesterol levels (10% and 44%, respectively). No significant effects on LDL or HDL cholesterol were observed. The gypenosides reduced nitrite approximately 80%. Similar results were obtained with atorvastatin (75 mg/kg for 4 days); except that LDL cholesterol was reduced (17%) and HDL cholesterol increased. 50% of lipoprotein lipase (LPL) plasma activity was inhibited by approximately 20 microM P407. Gynostemma had no effect on LL, however, it reversed the P407 inhibition of LPL activity in a concentration-dependent manner, with a 2-fold increase at approximately 10 microg/ml. CONCLUSIONS: These studies demonstrate efficacy of Gynostemma pentaphyllum in lowering triglyceride, cholesterol and nitrite in acute hyperlipidemia. The results suggest further investigations of Gynostemma gypenosides are warranted to examine the mechanisms of this activity. [Abstract/Link to Full Text]

Trommer H, Neubert RH
Screening for new antioxidative compounds for topical administration using skin lipid model systems.
J Pharm Pharm Sci. 2005;8(3):494-506.
PURPOSE: The effects of forty seven different substances (drugs, plant extracts, plant ingredients and polysaccharides) on UV irradiation induced lipid peroxidation were investigated. METHODS: Two lipid systems of different complexity were used as in vitro screening models. Iron ions were added as transition metal catalysts. A UV irradiation device was used to create high level radiation. The amount of lipid peroxidation secondary products was quantified by the thiobarbituric acid assay detecting malondialdehyde. RESULTS: The screening for antioxidative compounds for topical administration resulted in new, interesting findings. In the drug testings amantadine, bufexamac, tryptophan, melatonin, propranolol and hyaluronic acid were found to act antioxidatively whereas for ascorbic acid pro-oxidative effects were determined. Buckwheat extract significantly reduced the level of irradiation induced lipid peroxidation as well as the extracts of St. John's Wort, melissa and sage. The resistant starch novelose 330 and the samples of locust bean gum from a swing mill grinding series showed lipid protection after UV irradiation in the polysaccharide test rows. CONCLUSIONS: Human skin is constantly exposed to UV light and oxygen. Therefore, the administration of protectors in cosmetic formulations or sunscreens, as found in this study, may be helpful for the protection of the human skin against UV induced damage. In vivo experiments with substances found as protectors should follow to allow in vitro-in vivo correlation and clinical interpretation of the data. [Abstract/Link to Full Text]

Bhadra D, Bhadra S, Jain NK
Pegylated lysine based copolymeric dendritic micelles for solubilization and delivery of artemether.
J Pharm Pharm Sci. 2005;8(3):467-82.
PURPOSE: A newer polymeric amphiphilic micellar system was developed in the present study for solubilization and controlled delivery of an antimalarial drug, Artemether (ART). Methoxy polyethylene glycol (MPEG) 2000 and 5000 were used as hydrophilic terminal. METHODS: The hydrophobic di-Fluorene methoxycarbonyl-l-lysine (di-FMOC-L-lysine) was linked initially to the single reactive end of MPEG, and to the two amino groups of l-lysine by consecutive peptide linkages and deprotection upto 2.5 generations (G). Half-generations are diFMOC-lysine terminated systems and full-generations are deprotected l-lysine terminated systems. The half-generation (0.5 G, 1.5 G and 2.5 G) dendritic micelles of MPEG 2000 and 5000 were used to solubilize artemether. IR, NMR and Mass spectroscopy characterized the synthesis of these micellar systems. The CMC of the systems was determined. Then formulations made were characterized for solubility enhancement (i.e. drug loading) and drug-release profile. RESULTS: There is considerable solubility enhancement of artemether upto three to fifteen times depending on concentration, generation and type of dendritic micelles used. The size and shape were studied using transmission electron microscopy. The stability of the micellar formulation was also determined by storing the micelles at various temperatures for a definite period of time followed by its successive dilutions. The dendritic carriers were found to form stable micelles at 10-30 microg/ml (lower CMCs) depending on generation and type of MPEG used. The formulations increased the stability of the drug and also prolonged the release of artemether upto 1-2 days in vitro. CONCLUSION: From all the studies performed, it can be concluded that these micellar systems can be used for the safe and effective delivery of insoluble bioactive. [Abstract/Link to Full Text]

Sriram D, Yogeeswari P, Kumar TG
Microwave-assisted synthesis and anti-YFV activity of 2,3-diaryl-1,3-thiazolidin-4-ones.
J Pharm Pharm Sci. 2005;8(3):426-9.
PURPOSE: The purpose of this study was to prepare several 1,3-thaizolidin-4-ones bearing variously substituted diaryl ring at C-2 and N-3 positions and evaluate them for their anti-YFV activity. METHODS: Several 1,3-thaizolidin-4-ones were prepared by reacting substituted benzaldehyde with equimolar amount of an appropriate substituted aromatic amine in the presence of an excess of mercaptoacetic acid in toluene utilizing microwave irradiation. The synthesized compounds were also evaluated for their inhibitory effects on the replication of YFV in green monkey kidney (Vero) cells (ATCC CCL81), by means of a cytopathic effect reduction assay. RESULTS: The compound DS1 emerged as the most potent anti-YFV agent with EC50 of 6.9 microM and CC50 more than 100 microM making it more potent than ribavirin. CONCLUSION: 2,3-diaryl-1,3-thiazolidin-4-ones possess anti-YFV potency. [Abstract/Link to Full Text]

Almasirad A, Tajik M, Bakhtiari D, Shafiee A, Abdollahi M, Zamani MJ, Khorasani R, Esmaily H
Synthesis and analgesic activity of N-Arylhydrazone derivatives of mefenamic acid.
J Pharm Pharm Sci. 2005;8(3):419-25.
PURPOSE: A series of N-Arylhydrazone derivatives of mefenamic acid (a known non-steroidal anti-inflammatory drug) were synthesized in order to obtain new compounds with potential analgesic and anti-inflammatory activity. METHODS: The structures of all synthesized compounds were confirmed by means of infrared, proton magnetic resonance and mass spectroscopy. All compounds were evaluated for their analgesic and anti-inflammatory activities by abdominal constriction test (writhing test) and carrageenan-induced rat paw edema test respectively. RESULTS: Most of the synthesized compounds induced significant reduction in the writhing response when compared to control. Among them, compounds 11, 12, 15, 16, 19, 20, and 21 were significantly more potent than mefenamic acid in the writhing test. The anti-inflammatory activity of these 7 compounds were evaluated and compounds 11, 12, 16, 19 and 20 showed significant anti-inflammatory activity in comparison to control but their effect was weaker than mefenamic acid. CONCLUSIONS: The antinociceptive relative activity of some of these newly synthesized compounds is greater than mefenamic acid but they are not potent anti-inflammatory agents. [Abstract/Link to Full Text]

Skiba M, Bounoure F, Barbot C, Arnaud P, Skiba M
Development of cyclodextrin microspheres for pulmonary drug delivery.
J Pharm Pharm Sci. 2005;8(3):409-18.
PURPOSE: Microparticles of diameter < 5 microm were synthesized by interfacial cross-linking of 7.5% (w/v) beta-cyclodextrins (beta-CD) with 4.5% (w/v) terephtaloyle chloride in 1 M NaOH, in order to provide stable vector for drug encapsulation suitable for administration at the alveolar scale. METHODS: Batches were prepared varying different parameters such as amount of monomer (beta-CD) (5-30% w/v), NaOH concentration (0.5-4 M), reaction time (15-240 min), agitation rate (8000-24000 rpm), amount of cross-linking agent (terephtaloyle chloride: 1.25-10% w/v), surfactant percentage (2.5-10% of Span 85), studying the influence of the freeze-drying step. Microparticles were controlled with respect to their size by a laser diffraction technique, pH of the colloidal suspension, IR spectroscopy, Differential Scanning Calorimetry. After optimization of the microparticles size, complexation with amikacin sulfate was investigated comparing encapsulation efficiency and yield at each step of the preparation (solubilization, emulsification, cross-linking, freeze-drying), contact time and influence of the amount of amikacin. RESULTS: An optimized method was obtained with 1 M NaOH, 4.5% (w/v) cross-linking agent and 5% (w/v) surfactant agent, a 30 min reaction time, a 24000 rpm agitation rate, conducting to microparticles whose size is inferior to 5 microm. Amikacin sulfate encapsulation in polycondensed beta-cyclodextrin showed that better incorporation was obtained during the solubilization step or just before freeze-drying. CONCLUSIONS: Amikacin encapsulation in 5 microm diameter microparticles of beta-CD is achievable for pulmonary drug delivery. [Abstract/Link to Full Text]

Looije NA, Risovic V, Stewart DJ, Debeyer D, Kutney J, Wasan KM
Disodium Ascorbyl Phytostanyl Phosphates (FM-VP4) reduces plasma cholesterol concentration, body weight and abdominal fat gain within a dietary-induced obese mouse model.
J Pharm Pharm Sci. 2005;8(3):400-8.
PURPOSE: The purpose of this study was to determine if Disodium Ascorbyl Phytostanol Phosphates (FM-VP4) alters animal body weight and plasma lipid levels in a dietary-induced obese mouse model. METHODS: Twenty-four C57BL6 mice (28 days old) were housed individually and fed a standard mouse diet for 2 weeks upon arrival. After 2 weeks the animals were weighed and divided in 4 groups of similar average weight, and the groups received a low fat (10% kcal from fat) and high fat (45% kcal from fat) diet with or without FM-VP4 (2% w/w) for 12 continuous weeks. Food, water and caloric intake and body weight were recorded on a daily basis throughout the duration of the study. Following the 12th week of the study all animals were humanely sacrificed and blood and abdominal fat pads were harvested for further analysis. Plasma cholesterol, triglyceride, AST/ALT and creatinine levels were measured using enzymatic kits. RESULTS: There is a significant difference in weight gain between the low-fat diet and the low-fat diet + 2% w/w FM-VP4 treatment groups (P<0.05), as well as between the high-fat diet and the high-fat diet + 2% w/w FM-VP4 treatment groups (P<0.05). However, the reduction of weight gain of the high-fat diet + 2% FM-VP4 treatment group compared to the high-fat group was 51%, while the reduction in weight gain between the low-fat diet + 2% w/w FM-VP4 treatment group and the low-fat diet group was 17% over the duration of the study. No significant differences in food and water intakes, serum creatinine and AST/ALT levels were observed between the four groups. No significant differences in caloric intake between the low-fat diet and the low-fat diet + 2% w/w FM-VP4. However, a significant difference in caloric intake between high-fat diet and the high-fat diet + 2% w/w FM-VP4 treatment groups was observed. In addition, significant reductions in plasma cholesterol levels and abdominal fat pad weight between diet alone and diet + FM-VP4 treatment groups were observed. CONCLUSIONS: These findings suggest that FM-VP4 may have potential weight-loss and cholesterol lowering activity in both High Fat and Low Fat Diets treated groups. [Abstract/Link to Full Text]

Hosseinzadeh H, Sadeghnia HR
Safranal, a constituent of Crocus sativus (saffron), attenuated cerebral ischemia induced oxidative damage in rat hippocampus.
J Pharm Pharm Sci. 2005;8(3):394-9.
Increased oxidative stress has been implicated in the mechanisms of delayed neuronal cell death following cerebral ischemic insult. In this study, we investigated whether safranal, an active constituent of Crocus sativus L. stigmas, may ameliorate ischemia-reperfusion injury (IRI)-induced oxidative damage in rat hippocampus. Male NMRI rats were divided into six groups, namely, sham, control, ischemia and ischemia treated with safranal (four groups). The transient global cerebral ischemia was induced using four-vessel-occlusion method for 20 min. Safranal was injected intraperitoneally (727.5 mg/kg, 363.75 mg/kg, 145.5 mg/kg, and 72.75 mg/kg body weight) 5 min. prior to reperfusion and the administration was continued every 24 hours for 72 hours after induction of ischemia. The markers of oxidative stress including thiobarbituric acid reactive substances (TBARS), total sulfhydryl (SH) groups and antioxidant capacity of hippocampus (using FRAP assay) were measured. The transient global cerebral ischemia induced a significant increase in TBARS levels (p<0.001), decrement in both antioxidant power (FRAP value) (p<0.05) and total sulfhydryl (SH) concentrations (p<0.001) in comparison with sham-operated animals. Following safranal administration the total SH contents (3.2 vs. 0.7micromol/g, p<0.001, safranal 727.5 mg/kg) and antioxidant capacity (4.12 vs. 1.16 micromol/g, p<0.001; 727.5 mg/kg) were elevated in hippocampus in comparison with ischemic group. The MDA level was declined significantly in hippocampus (52.31 vs. 159.70 nmol/g, p<0.001; 727.5 mg/kg). It is concluded that safranal have some protective effects on different markers of oxidative damage in hippocampal tissue from ischemic rats. [Abstract/Link to Full Text]

Hosseinzadeh H, Sadeghnia HR, Ziaee T, Danaee A
Protective effect of aqueous saffron extract (Crocus sativus L.) and crocin, its active constituent, on renal ischemia-reperfusion-induced oxidative damage in rats.
J Pharm Pharm Sci. 2005;8(3):387-93.
PURPOSE: The generation of reactive oxygen species and lipid peroxidation are associated with tissue injury following ischemic insult; therefore, the use of antioxidants appears rational in the improvement of kidney diseases therapy. The aim of the present study was to assess the effect of aqueous saffron extract (Crocus sativus L.) and its active constituent, crocin, on oxidative stress following renal ischemia-reperfusion injury (IRI) in rats. METHODS: The cellular redox status (thiobarbituric acid reactive species (TBARS) and total thiol levels) and antioxidant power (using ferric reducing/antioxidant power test) were assessed in control and ischemic groups. The left kidney was exposed to warm ischemia for 60 min followed by reperfusion for 90 min. The macerated aqueous extract of saffron (with doses of 5, 20 and 80 mg/kg, i.p.) and crocin (with doses of 50, 200 and 400 mg/kg, i.p.) were administrated prior to induction of ischemia. Normal saline (10 ml/kg, i.p.) was injected to control group and a sham group that did not have ischemia-reperfusion. RESULTS: Ischemia-reperfusion (IR) caused a significant increase in TBARS levels (p<0.001) and decrement in both antioxidant power (FRAP value) (p<0.05) and total thiol concentration (p<0.001) in kidney homogenate samples. In crocin pretreated groups, a reduction in TBARS levels (from 85.8 +/- 5.4 to 20.9 +/- 1.5 nmol/g tissue, p<0.001; 400 mg/kg) and elevation in antioxidant power (FRAP value) (from 3.05 +/- 0.16 to 4.15 +/- 0.16 micromol/g tissue, p<0.001; 400 mg/kg) and total thiol concentrations (from 0.38 +/- 0.03 to 0.62 +/- 0.03 mM, p<0.001; 200 mg/kg), as compared with control group, were observed. The aqueous extract also reduced lipid peroxidation products (from 85.8 +/- 5.4 to 15.9 +/- 2.6 nmol/g tissue, p<0.001; 80 mg/kg) and increased antioxidant power (from 2.98 +/- 0.11 to 5.97 +/- 0.56 micromol/g tissue, p<0.001; 80 mg/kg) in ischemia-reperfusion injured rat kidneys. CONCLUSION: This study therefore suggests that the aqueous saffron extract (Crocus sativus L.) and its active constituent, crocin, may be useful agents for the prevention of renal ischemia-reperfusion (IR)-induced oxidative injury in rats. [Abstract/Link to Full Text]

Roupe KA, Helms GL, Halls SC, Y��ez JA, Davies NM
Preparative enzymatic synthesis and HPLC analysis of rhapontigenin: applications to metabolism, pharmacokinetics and anti-cancer studies.
J Pharm Pharm Sci. 2005;8(3):374-86.
PURPOSE: A facile method was established to enzymatically synthesize rhapontigenin from the glycosylated parent compound rhaponticin. A novel and simple high-performance liquid chromatographic method was developed for the determination of rhapontigenin. The assay was successfully applied to both the in vitro and in vivo metabolic kinetic study of rhapontigenin. METHODS: Serum, or microsomes (0.1 mL) was precipitated with acetonitrile after addition of the internal standard, daidzein. Separation was achieved on an amylose tris 3,5 dimethylphenylcarbamate column (150 x 4.6 mm, ID, 5m) with UV detection at 324 nm. Hep G2 hepatoma cells were treated with rhapontigenin or rhaponticin (0-250 microg/mL) and cell viability was measured. RESULTS: The calibration curves were linear ranging from 0.5 to 100 micromg/mL. The mean extraction efficiency was > 99%. Precision of the assay (coefficient of variation) was < 5%, including the limit of quantitation (0.5 microg/mL). Bias of the assay was lower than 5%. The limit of detection was 100 ng/mL for a 0.1 mL sample. One glucuronidated metabolite of rhapontigenin has been identified. Preliminary pharmacokinetic data revealed the presence of a glucuronidated metabolite in the serum and a terminal elimination t1/2 of approximately 6 h. Rhapontigenin demonstrated concentration-dependent anti-cancer activity with an IC50 115 microg/mL in HEP G2 cells while rhaponticin showed no activity across the concentrations tested in vitro. CONCLUSIONS: The preparative enzymatic synthesis method has demonstrated utility to provide sufficient rhapontigenin for pharmaceutical studies. Rhapontigenin is an active anti-cancer compound. The developed HPLC assay is sensitive, reproducible and accurate and can be applied to pharmacokinetic and metabolism studies. [Abstract/Link to Full Text]

B�rger C, Fischer DR, Cordenunzzi DA, Batschauer AP, Cechinel Filho V, Soares AR
Acute and subacute toxicity of the hydroalcoholic extract from Wedelia paludosa (Acmela brasiliensis) (Asteraceae) in mice.
J Pharm Pharm Sci. 2005;8(2):370-3.
PURPOSE: The present study was carried out to evaluated acute and subacute toxicity of a hydroalcoholic extract from aerial parts of Wedelia paludosa (Asteraceae). METHODS: Toxicity of W. paludosa was evaluated in Swiss mice after ingestions of the extract during one day (acute model) and during 15 days (subacute model). RESULTS: The results showed that the LD50 of the extract is higher than 4000 mg/kg and the subacute treatment did not shows any change in corporal weight and hematological parameters. However, a change in liver weight but not in hepatic enzymes was observed. This suggests that the liver function is not altered by Wedelia paludosa in this study. Some changes in the creatinine content were observed, but could not be related with the extract dose. CONCLUSIONS: The results suggest that the plant seems to be destituted of toxic effects in mice. [Abstract/Link to Full Text]

Preechagoon D, Sumyai V, Tontisirin K, Aumpon S, Pongjanyakul T
Formulation development and stability testing of oral morphine solution utilizing preformulation approach.
J Pharm Pharm Sci. 2005;8(2):362-9.
PURPOSE: Prefomulation approach utilizing the fractional-ordered randomized blocked design was employed for the formulation development and stability testing of morphine solution. METHODS: Factors expecting to affect the stability of morphine were evaluated, i.e., vehicle, antioxidant, chelating agent, and pH of the solution. Eight formulations of a possible 16 were prepared according to the block design. The stability of the preparations was tested after 35 days of storage. The data of preformulation study were used for formulation development. RESULTS: The presence of glycerin and ethylenediamine-tetraacetic acid in the formulation, and the pH of the solution adjusted to 4, stabilized morphine. The concentration of morphine decreased drastically in the formulations containing sodium metabisulfite, and those pH adjusted to 6. After 35 days, only 65% of morphine was found in the formulation containing sodium metabisulfite and pH adjusted to 6. The results of preformulation study were used for preparing oral morphine preparations. Samples were kept in amber glass bottles and stored at 4 degrees C and 25 degrees C/75% RH for 13 months. No precipitation of the four formulations was detected. Only a decrease of odor and a small increase of pH value of the preparations (< 0.3 units) were observed. More than 97% of morphine remained in all samples. The samples were free from microbial contamination. CONCLUSION: Stable morphine solution formulations can be achieved with the utilization of the preformulation approach. They were stable more than 13 months when stored at 4 degrees C and 25 degrees C/75% RH. [Abstract/Link to Full Text]

Moridani MY, Moore M, Bartsch RA, Yang Y, Heibati-Sadati S
Structural toxicity relationship of 4-alkoxyphenols' cytotoxicity towards murine B16-F0 melanoma cell line.
J Pharm Pharm Sci. 2005;8(2):348-60.
PURPOSE: The aim of this study was to identify phenolic agents that could form quinone reactive intermediate metabolites in melanocytes in order to be effective as anti-melanoma agents; but were not metabolized by liver P450 metabolizing enzymes in order to have minimal toxicity towards the liver. METHODS: Tyrosinase, an enzyme present abundantly in melanocytes was selected as a molecular target for the treatment of malignant melanoma. Ten alkoxyphenols were investigated for their metabolism by tyrosinase/O2, rat liver P450 microsomal/NADPH/O2 metabolizing systems and for their toxicity towards B16-F0 melanoma cells. RESULTS: All the alkoxyphenols showed a dose- and time-dependent toxicity towards B16-F0 cells except 2-iso-propoxyphenol. 4-n-hexyloxyphenol demonstrated the greatest toxicity towards B16-F0 cells while minimally depleting glutathione in microsomal preparations at its calculated LC10 and LC50 lethal concentrations for B16-F0. At 100 microM concentrations, 4-t-butoxyphenol showed the lowest amount of glutathione depletion by microsomal P450 system. Alkoxyphenols with at least two alkyl groups derivatized at alpha carbon of alkoxy group showed minimal rates of metabolism by tyrosinase/O2 metabolizing system. A quantitative structural toxicity relationship equation was also derived, LogLC50(mM)= -0.265(+/-0.064)LogP + 2.482(+/-0.179). CONCLUSIONS: 4-n-hexyloxy-phenol was identified as a potential lead anti-melanoma agent against B16-F0 melanoma cells with minimal metabolism by rat liver P450 microsomal preparation. [Abstract/Link to Full Text]

Massaroti P, Cassiano NM, Duarte LF, Campos DR, Marchioretto MA, Bernasconi G, Calafatti S, Barros FA, Meurer EC, Pedrazzoli J
Validation of a selective method for determination of paroxetine in human plasma by LC-MS/MS.
J Pharm Pharm Sci. 2005;8(2):340-7.
PURPOSE: A sensitive, robust, and selective liquid chromatographic-tandem mass spectrometric method (LC-MS/MS) was developed and validated for paroxetine quantification in human EDTA plasma. METHODS: Sample preparation was based on liquid-liquid extraction using a mixture of ethyl acetate/hexane (50/50; v/v) to extract the drug and internal standard from plasma. Chromatography was performed on a C-18 analytical column and the retention times were 1.6 and 1.7 for paroxetine and fluoxetine (IS), respectively. The ionization was optimized using ESI(+) and selectivity was achieved by tandem mass spectrometric analysis using MRM functions, 330.0 --> 70.0 and 310 --> 43.9 for paroxetine and fluoxetine. RESULTS: Analytical curve ranged from 0.2 to 20.0 ng/mL. Inter-day precision and accuracy of the quality control (QC) samples were < 15% relative standard deviation (RSD). Analyte stability during sampling processing and storage were established. CONCLUSION: Validation results on linearity, specificity, accuracy, precision as well as application to the analysis of samples taken up to 120 h after oral administration of 20 mg of paroxetine in 28 healthy volunteers were found to be of good performance in bioequivalence study. [Abstract/Link to Full Text]

Malheiros A, Cechinel Filho V, Schmitt CB, Yunes RA, Escalante A, Svetaz L, Zacchino S, Delle Monache F
Antifungal activity of drimane sesquiterpenes from Drimys brasiliensis using bioassay-guided fractionation.
J Pharm Pharm Sci. 2005;8(2):335-9.
PURPOSE: This study describes the antifungal effect of extracts and compounds isolated from Drimys brasiliensis acting against dermatophytes. METHODS: The activities were evaluated by using the microbroth dilution method. RESULTS: Bioassay-guided fractionation of the most active extract from the bark (CHCl3) led to the isolation of the sesquiterpene drimanes polygodial, 1-beta-(p-methoxycinnamoyl)-polygodial, drimanial and 1-beta-(p-cumaroyloxy)-polygodial, which were selectively active against Epidermophyton floccosum and Tricophyton rubrum. CONCLUSIONS: The selective antifungal activity reported in this paper for drimanes isolated from D. brasiliensis opens the possibility that they could be helpful for the developing of new antifungal agents for treating the difficult to eradicate dermatomycoses produced by E. floccosum. [Abstract/Link to Full Text]

C�ceres-L�riga FM, P�rez-L�pez H, Santos-Gracia J, Morlans-Hern�ndez K, Marrero-Mirayaga MA
Thrombolytic treatment as first option in recurrent tricuspid prosthetic valve thrombosis and Ebstein's anomaly.
J Pharm Pharm Sci. 2005;8(2):332-4.
Ebstein's anomaly is the most frequent cause of congenital tricuspid regurgitation. The coexistence of a mechanical heart prosthesis in a low-pressure circuit and poor compliance in the anticoagulant therapy contributed decisively to the appearance of recurrent mechanical heart valve thrombosis in these patients. A 49 years old female patient is reported where thrombolytic therapy with recombinant Streptokinase (TT-rSK) was the first treatment choice in seven recurrent episodes of prosthetic valve thrombosis. [Abstract/Link to Full Text]

Awad A, Eltayeb I, Matowe L, Thalib L
Self-medication with antibiotics and antimalarials in the community of Khartoum State, Sudan.
J Pharm Pharm Sci. 2005;8(2):326-31.
PURPOSE: To estimate the prevalence of self medication with antibiotics and antimalarials in Khartoum State, Sudan and evaluate factors associated with self medication. METHODS: A pre-tested questionnaire was used to collect data from a sample of 600 households, (1750 adult persons), selected from three cities in Khartoum State, Sudan, using a multistage stratified clustered sampling. RESULTS: One thousand two hundred and ninety three (73.9%) of the study population had used antibiotics or antimalarials without a prescription within one month prior to the study. Eight hundred and forty one (48.1%) of the respondents agreed that they have used antibiotics, 43.4% used antimalarials, while 17.5% used both. Self medication with either antibiotics/ antimalarials was found to be significantly associated with age, income, gender and level of education. Overall, self medication with any antibiotics or antimalarials was least common among the > or = 60 years compared to youngest age group (OR: 0.07; 0.04 -0.11) and most common among the female gender (OR: 1.8; 1.4 -2.4), the middle income group (OR: 3.7; 2.6-5.3) and the university graduates. Self medication with antibiotic was found to be significantly higher among females (OR: 1.5; 1.16-1.87), middle aged respondents aged 40-59 (OR: 2.1; 1.5-3.0) compared to younger respondents. Lower income and higher level of education was also found to be significantly associated with the increase risk of self medicating with antibiotic. Increase risk for self medication with antimalarials were, however, found to be significantly associated with male gender and younger age group of < 40 years and middle income earners and less educated respondents. The main reason that was indicated for the self-medication was financial constraints. The main source of medicines was the private pharmacies, which were regarded as a cheaper alternative to other primary healthcare sources. CONCLUSION: The prevalence of self-medication with antibiotics/antimalarials in Khartoum State, Sudan is alarmingly high. Self medication behaviour varies significantly with a number of socio-economic characteristics. Given the growing global resistance for antibiotic and documented health issues related to inappropriate use of such drugs, our findings has major public health policy implications for countries like Sudan. [Abstract/Link to Full Text]

Chiadmi F, Lyer A, Cisternino S, Toledano A, Schlatter J, Ratiney R, Fontan JE
Stability of levamisole oral solutions prepared from tablets and powder.
J Pharm Pharm Sci. 2005;8(2):322-5.
PURPOSE: To study the stability of levamisole oral solutions (25 mg/mL) prepared from powder and tablets stored at 4 +/- 3 degrees C and 23 +/- 2 degrees C in amber glass prescription bottles. METHODS: Levamisole 25 mg/mL solutions were prepared from commercially available 50-mg tablets or from pure powder in sterile water. Levamisole concentrations were determined in duplicate by a stability-indicating HPLC method at 0, 1, 2, 3, 4, 7, 14, 30, 60 and 90 days. The initial and final pHs of solutions were measured. RESULTS: The recovery of levamisole from tablets was 100 +/- 2.1%. No color or odour changes were observed during the study period. The oral solutions prepared from powder were stable at least 90 days stored at 4 and 23 degrees C. The oral solutions prepared from tablets were stable at least 90 days at 4 degrees C and 15 days when stored at 23 degrees C. The initial pH of solutions prepared from powder and tablets were 5.30 and 4.55, respectively. Initial and final pH values were significantly different (p<0.001) for the two solutions. CONCLUSIONS: Levamisole 25 mg/mL oral solutions can be prepared from tablets or powder with sterile water for irrigation and stored for 90 days under refrigeration, taking account of the lack of microbiological contamination. [Abstract/Link to Full Text]

Dakshayani KB, Subramanian P, Manivasagam T, Essa MM, Manoharan S
Melatonin modulates the oxidant-antioxidant imbalance during N-nitrosodiethylamine induced hepatocarcinogenesis in rats.
J Pharm Pharm Sci. 2005;8(2):316-21.
PURPOSE: Melatonin, the principle hormone of pineal gland plays an important role in several biological processes. The effects of melatonin on hepatic marker enzymes [aspartate and alanine transaminases (AST and ALT)], lipid peroxides [thiobarbituric acid reactive substances (TBARS)] and antioxidants [reduced glutathione (GSH), glutathione peroxidase (GPx) and glutathione-S-transferase (GST)] during N-nitrosodiethylamine (NDEA)-induced hepatocarcinogenesis in rats were studied. METHODS: Male albino Wistar rats of body weight 150-170 g were divided into four groups of six animals each. Group I animals served as control, Group II animals received single intraperitoneal injection of NDEA at a dose of 200 mg/kg body weight followed by weekly subcutaneous injections of CCl4 at a dose of 3 mL/kg body weight. Group III animals were treated as in Group II and melatonin (5 mg/kg body weight) was administered intraperitoneally. Group IV animals received melatonin alone at the same dose as Group III animals. RESULTS: A significant increase in the activities of serum AST and ALT was observed in NDEA treated rats when compared with control animals. Melatonin administered rats showed a significant decrease in the activities of these enzymes when compared with NDEA treated animals. In the liver of NDEA-treated animals, decreased lipid peroxidation associated with enhanced antioxidant levels was observed. Administration of melatonin positively modulated these changes. CONCLUSION: Our results indicate that melatonin exerts chemopreventive effect by restoring the activities of hepatic marker enzymes and reversing the oxidant-antioxidant imbalance during NDEA-induced hepatocarcinogenesis. [Abstract/Link to Full Text]

Tirnaksiz F, Kalsin O
A topical w/o/w multiple emulsions prepared with Tetronic 908 as a hydrophilic surfactant: formulation, characterization and release study.
J Pharm Pharm Sci. 2005;8(2):299-315.
PURPOSE: The aim of this work was to prepare the stable water/oil/water multiple emulsions (MEs), to investigate the usage of poloxamine 908, to observe the influence of surfactant percentage on the properties of MEs. METHOD: MEs were prepared by liquid paraffin, cetyl dimethicone copolyol and poloxamine 908 by a two-step emulsification procedure. Caffeine was used as a water-soluble model. The viscosity, conductivity and globule size of MEs were followed over time. RESULTS: The formulations containing 1% cetyl dimethicone copolyol and 1 or 2% poloxamine 908 were the most stable systems. The globule size of MEs ranged from 20 to 37 microm and did not change during time. The yield of MEs was between 99.6% and 98.7%. The conductivity increased and the viscosity of systems decreased during time. Increase in poloxamine 908 influenced the viscosity of the system, with the viscosity decreasing as the hydrophilic surfactant concentrations were increased. Caffeine release from the MEs was slow; the release was affected by both surfactant concentrations. CONCLUSION: Poloxamine 908 could be used as a hydrophilic surfactant for formulation of w/o/w MEs. The concentration of poloxamine 908 was a very important parameter in preparing stable MEs. It was concluded that caffeine might be transported out by molecular diffusion and through a reverse micellar mechanism controlled by the viscosity of the system. [Abstract/Link to Full Text]

Breier AR, Paim CS, Steppe M, Schapoval EE
Development and validation of dissolution tests for fexofenadine hydrochloride capsules and coated tablets.
J Pharm Pharm Sci. 2005;8(2):289-98.
PURPOSE: This study describes the development and validation of dissolution tests for fexofenadine hydrochloride capsules and coated tablets using an HPLC method. METHOD: The appropriate conditions were determinate after testing sink conditions, dissolution medium, and agitation intensity. The apparatus, paddle and basket, were applied to tablets and capsules, respectively. Fexofenadine hydrochloride capsules, products A and B, and coated tablets, products A, B and C were evaluated. The best dissolution conditions tested, for the products in each respective pharmaceutical dosage form were applied to evaluate the dissolution profiles. The parameters of difference factor, similar factor, and dissolution efficacy were employed. RESULTS: Optimal conditions to carry out the dissolution tests were 900 ml of 0.01 M hydrochloric acid as dissolution medium, basket at 100 rotation per minute (rpm) stirring speed for capsules and paddle at 75 rpm for tablets. The dissolution profiles for tablets products A, B, and C and for capsules products A and B were not similar. CONCLUSION: The developed and validated dissolution tests satisfactorily describes the time-course of the drug release. The obtained results provided adequate dissolution profiles. The HPLC method was validated to quantify fexofenadine capsules and coated tablets from the dissolution tests. [Abstract/Link to Full Text]

St-Onge L, Archambault JF, Kwong E, Sabsabi M, Vadas EB
Rapid quantitative analysis of magnesium stearate in tablets using laser-induced breakdown spectroscopy.
J Pharm Pharm Sci. 2005;8(2):272-88.
PURPOSE: Laser-induced breakdown spectroscopy (LIBS) was evaluated for its potential as a process analytical tool for the rapid determination of magnesium stearate (MgSt) distribution within and between tablets as well as between batches in a typical manufacturing run, and for the comparison of direct-compression and roller-compaction processes. METHODS: These studies were conducted using a prototype instrument and a commercial PharmaLIBS unit, both based on pulsed Nd:YAG laser radiation at 1064 nm. The intensity of a magnesium spectral line either at 517.27 or 518.36 nm was used, depending on the product, to quantitate the MgSt concentration in the tablets. RESULTS: Using internal standardization, it was possible with the prototype instrument to accurately quantitate MgSt at the 0.5% level in two different products. For eight batches of one product, using 10 tablets from each batch, the intra-tablet, intra-batch, and inter-batch MgSt %RSDs were found to be 13.8%, 5.4% and 7.4%, respectively. Further studies were conducted with the commercial LIBS unit, which showed similar performance as the prototype unit. In particular, it was found that different depth-profile distributions of MgSt were associated with roller-compacted tablets and direct-compressed tablets. CONCLUSION: These findings illustrate the potential of LIBS to be developed as a process analytics tool for the direct and rapid determination of MgSt content and distribution in tablets. [Abstract/Link to Full Text]

Wasan KM, Looije NA
Emerging pharmacological approaches to the treatment of obesity.
J Pharm Pharm Sci. 2005;8(2):259-71.
The obesity epidemic has been recognized by the World Health Organization (WHO) as one of the top 10 global health problems. Worldwide, more than one billion adults are overweight and over 300 million are obese. The majority of developed countries, including the United States, Canada and England are experiencing dramatic increases in obesity. Obesity is a condition associated with the accumulation of excessive body fat resulting from chronic imbalance of energy whereby the intake of energy exceeds expenditure. The excess body fat predisposes an obese individual to chronic diseases, such as coronary heart disease, type 2 diabetes and diseases of the gall bladder and cancer. The high incidence of obesity and the lack of safe pharmaceutical agents have fuelled an increase in anti-obesity drug-related research. Although a number of pharmacological approaches have been investigated in recent years, few safe, therapeutically effective products have been developed. This commentary focuses on emerging pharmacological approaches targeted for the treatment of obesity. [Abstract/Link to Full Text]

Arulsudar N, Subramanian N, Muthy RS
Comparison of artificial neural network and multiple linear regression in the optimization of formulation parameters of leuprolide acetate loaded liposomes.
J Pharm Pharm Sci. 2005;8(2):243-58.
PURPOSE: We planned to optimize the effect of formulation variables on the percent drug entrapment (PDE) of the liposomes encapsulating leuprolide acetate by reverse phase evaporation method using Artificial neural network (ANN) and Multiple linear regression (MLR). METHOD: Twenty seven formulations were prepared based on 3x3 factorial design. The volume of aqueous phase (X(1)), HSPC/DSPG [negative charge] (X(2)), and HSPC/Cholesterol (X(3)) were selected as the causal factors. Potential variables such as concentration of lipid: drug and hydration medium were kept constant in experimental design. The PDE (dependent variable) and the transformed values of independent variables were subjected to multiple regression analysis to establish a second order polynomial equation (full model). A set of PDE and causal factors was used as tutorial data for the ANN and fed into a computer. The feed forward back propagation (bp) method was optimized. The ANN model and MLR were validated for accurate prediction of PDE. RESULTS: To simplify the polynomial equation, F-statistic was applied to reduce polynomial equation (reduced model) by neglecting non-significant (P<0.05) terms. The reduced polynomial equation was used to plot three two-dimensional contour plots at fixed levels of -1, 0 and 1 of the variable X(3) to obtain various combination values of the two other independent variables (X(1) and X(2)) at predetermined PDE. The root mean square value of the trained ANN model by feed forward bp method was 0.0000354, which indicated that the optimal model was reached. The optimization methods developed by both ANN and MLR were validated by preparing another six liposomal formulations. The predetermined PDE (from ANN and MLR) and the experimental data were compared with predicted data by paired "t" test, no statistically significant difference was observed. ANN showed less error compared to MLR. CONCLUSIONS: These findings demonstrate that the ANN model provides more accurate prediction and is quite useful in the optimization of pharmaceutical formulations when compared to multiple regression analysis method. The normalized error (NE) value observed with the optimal ANN model was 0.0211 while it was 0.0658 for the full model in the case of second-order polynomial equation composed of the combination of causal factors (X(1), X(2) and X(3)). Thus the derived equation, contour plots and ANN helps in predicting the values of the independent variables for maximum PDE in the preparation of leuprolide acetate liposomes by reverse phase evaporation technique. [Abstract/Link to Full Text]

Yusuff KB, Balogun O
Physicians' prescribing of anti-hypertensive combinations in a tertiary care setting in southwestern Nigeria.
J Pharm Pharm Sci. 2005;8(2):235-42.
PURPOSE: To evaluate physicians' prescribing of anti-hypertensive drug combinations in a tertiary care setting in southwestern Nigeria, determine the degree of usage of Angiotensin Converting Enzyme (ACE) inhibitor-based combinations and identify specific points of intervention to improve outcomes of anti-hypertensive combination therapy. METHODS: A cross-sectional retrospective drug use review was conducted between June 1st and August 31st 2002 using randomly selected 200 case notes of patients attending the Hypertension Clinic at a 900-bed tertiary care facility in southwestern Nigeria. 11 case notes were not used due to incompleteness. RESULTS: 73% (138) of the patients were on anti-hypertensive drug combinations, comprising 71.7% (99), 24.4% (34) and 3.6% (5) on combinations of two, three and four drugs respectively. Overall, Thiazide diuretic consisting mainly of fixed dose combination of Amiloride and Hydorchlorothiazide (Moduretic(r)) was the most frequently prescribed drug class in anti-hypertensive combination therapy (83.3%). ACE inhibitor, Lisinopril (Zestril(r)), was prescribed in combination with Moduretic(r), Calcium channel blocker and beta-blocker in 6.5%, 8.5% and 0.7% respectively. Blood pressure control was adequate in only 29% (40) of patients, though adherence with therapy was documented as adequate in 77.5% (107). Type-2 diabetes mellitus (32.7%) and osteoarthritis (21.8%) were the most frequent co-morbidities. Potentially harmful drug-drug interactions in the study sample were identified in 17.5% (46) of patients. Physician documentation of adverse drug reactions among patients was done in only 10.9% of cases. There appear to be no institutionalised system in place to monitor, detect and document adverse drug reactions among patients on anti-hypertensive drug therapy. CONCLUSION: Physicians' prescribing of anti-hypertensive drug combinations in a tertiary care setting in southwestern Nigeria is considerable. However, this practice does not appear to have positively impacted on blood pressure control among hypertensive patients nor being modulated by an Institutionalised standard guide. [Abstract/Link to Full Text]

Berruet N, Sentenac S, Auchere D, Gimenez F, Farinotti R, Fernandez C
Effect of efavirenz on intestinal p-glycoprotein and hepatic p450 function in rats.
J Pharm Pharm Sci. 2005;8(2):226-34.
PURPOSE: P-glycoprotein (P-gp) and cytochrome P450 (P450) affect drug disposition. Efavirenz (EFV) is an anti-HIV drug used in combination. Since most anti-HIV medications are substrate and modulators of P-gp and/or P450, we investigated the effects of EFV on intestinal P-gp and hepatic P450 function to predict drug interactions. METHODS: (i) The effect of EFV on rat intestinal P-gp function was studied on everted gut sacs and in situ intestinal perfusion. EFV was orally administered (150 mg/kg) for 6 days. Then, rhodamine 123 was used as a P-gp substrate and verapamil as an inhibitor. P-gp function was evaluated by the difference between rhodamine 123 transport with and without verapamil. (ii) The effect of EFV on rat hepatic P450 metabolism was investigated using hepatic microsomes, prepared from rats pretreated or not with EFV. RESULTS : In everted gut sacs, P-gp function was not modified and in the in situ intestinal perfusion, rhodamine 123 clearance was not affected by EFV. Concentrations of the metabolites, 1-OH midazolam and 4-OH midazolam were higher in EFV pretreated rats than those in the control group. CONCLUSION: EFV should not modify intestinal absorption of co-administered substrates of P-gp, but could decrease plasma concentrations of co-administered drugs metabolized by P450. [Abstract/Link to Full Text]

Emmerton L, Marriott J, Bessell T, Nissen L, Dean L
Pharmacists and prescribing rights: review of international developments.
J Pharm Pharm Sci. 2005;8(2):217-25.
PURPOSE: Continuity of care, equitable access, and quality and safety are major foci in health services management. The introduction of limited prescribing rights to pharmacists has the potential to reduce fragmentation within the health system, optimise medication management, improve continuity of patient care and improve patient access to medication. RESULTS: Eight models for pharmacists' prescribing have been implemented internationally, varying in their dependency on protocols, formularies and collaboration with physicians. These have also been described using terms such as Supplementary Prescribing and Patient Group Directions. CONCLUSION: Issues relating to practical implementation of pharmacists' prescribing include negotiation of national health policy, pharmacists' training and accreditation, liability, reimbursement and documentation. [Abstract/Link to Full Text]

Recent Articles in Journal of Drug Targeting

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Recent Articles in Drug Metabolism and Pharmacokinetics

Imai T
Human carboxylesterase isozymes: catalytic properties and rational drug design.
Drug Metab Pharmacokinet. 2006 Jun;21(3):173-85.
Human carboxylesterase 1 (hCE-1, CES1A1, HU1) and carboxylesterase 2 (hCE-2, hiCE, HU3) are a serine esterase involved in both drug metabolism and activation. Although both hCE-1 and hCE-2 are present in several organs, the hydrolase activity of liver and small intestine is predominantly attributed to hCE-1 and hCE-2, respectively. The substrate specificity of hCE-1 and hCE-2 is significantly different. hCE-1 mainly hydrolyzes a substrate with a small alcohol group and large acyl group, but its wide active pocket sometimes allows it to act on structurally distinct compounds of either large or small alcohol moiety. In contrast, hCE-2 recognizes a substrate with a large alcohol group and small acyl group, and its substrate specificity may be restricted by a capability of acyl-hCE-2 conjugate formation due to the presence of conformational interference in the active pocket. Furthermore, hCE-1 shows high transesterification activity, especially with hydrophobic alcohol, but negligible for hCE-2. Transesterification may be a reason for the substrate specificity of hCE-1 that hardly hydrolyzes a substrate with hydrophobic alcohol group, because transesterification can progress at the same time when a compound is hydrolyzed by hCE-1. From the standpoint of drug absorption, the intestinal hydrolysis by CES during drug absorption is evaluated in rat intestine and Caco2-cell line. The rat in situ single-pass perfusion shows markedly extensive hydrolysis in the intestinal mucosa. Since the hydrolyzed products are present at higher concentration in the epithelial cells rather than blood vessels and intestinal lumen, hydrolysates are transported by a specific efflux transporter and passive diffusion according to pH-partition. The expression pattern of CES in Caco-2 cell monolayer, a useful in vitro model for rapid screening of human intestinal drug absorption, is completely different from that in human small intestine but very similar to human liver that expresses a much higher level of hCE-1 and lower level of hCE-2. Therefore, the prediction of human intestinal absorption using Caco-2 cell monolayers should be carefully monitored in the case of ester and amide-containing drugs such as prodrugs. Further experimentation for an understanding of detailed substrate specificity for CES and development of in vitro evaluation systems for absorption of prodrug and its hydrolysates will help us to design the ideal prodrug. [Abstract/Link to Full Text]

Tsujimoto M, Hirata S, Dan Y, Ohtani H, Sawada Y
Polymorphisms and linkage disequilibrium of the OATP8 (OATP1B3) gene in Japanese subjects.
Drug Metab Pharmacokinet. 2006 Apr;21(2):165-9.
OATP8, a member of the organic anion-transporting polypeptide family, is expressed on the sinusoidal membrane of hepatocytes, and transports endogenous organic anions, such as 17beta-glucuronosyl estradiol, and xenobiotic substances, such as digoxin. The objective of this study is to search for polymorphisms of the OATP8 gene and to assess the allele frequency of the polymorphisms in the Japanese population. Analysis of the OATP8 gene in 79 subjects revealed complete linkage of two deletion polymorphisms in the 5' regulatory region, deletion from position -28 to -11 and deletion from position -7 to -4, with an allele frequency of 0.196 for the deletion allele. The polymorphisms T334G (Ser112Ala) and G699A (Met233Ile) were also shown to be in complete linkage disequilibrium, with an allele frequency of 0.728 for the variant (112Ala/233Ile) allele. Interestingly, linkage disequilibrium was identified between the ins/del polymorphism and SNPs at 112 and 233. The predicted major haplotype was the insert-variant type with a haplotype frequency of 0.60. [Abstract/Link to Full Text]

Taguchi M, Urai M, Taira S, Tanabe H, Hashimoto Y
Endogenous uremic substances are not involved in the reduced hepatic extraction of metoprolol in bilateral ureter-ligated rats.
Drug Metab Pharmacokinet. 2006 Apr;21(2):156-64.
The hepatic extraction of metoprolol is reduced in rats with bilateral ureter ligation (BUL)-induced renal failure. The aim of the present study was to evaluate the effect of uremic substances on the hepatic metabolism of metoprolol in rats with BUL. The metabolic rate in the liver microsomes of BUL rats was similar to that in sham rats, and there was no significant difference between sham and BUL rats in the effect of the supernatant of liver homogenates on the metabolism. The rate of metabolism in the liver microsomes in the presence of the plasma of BUL rats was also similar to that in the presence of the plasma of sham rats. These findings indicated that uremic substances which accumulate in BUL rats do not directly inhibit the activity of CYP2D2, which is responsible for the metabolism of metoprolol in the rat liver. [Abstract/Link to Full Text]

Yamaori S, Fujiyama N, Kushihara M, Funahashi T, Kimura T, Yamamoto I, Sone T, Isobe M, Ohshima T, Matsumura K, Oda M, Watanabe K
Involvement of human blood arylesterases and liver microsomal carboxylesterases in nafamostat hydrolysis.
Drug Metab Pharmacokinet. 2006 Apr;21(2):147-55.
Metabolism of nafamostat, a clinically used serine protease inhibitor, was investigated with human blood and liver enzyme sources. All the enzyme sources examined (whole blood, erythrocytes, plasma and liver microsomes) showed nafamostat hydrolytic activity. V(max) and K(m) values for the nafamostat hydrolysis in erythrocytes were 278 nmol/min/mL blood fraction and 628 microM; those in plasma were 160 nmol/min/mL blood fraction and 8890 microM, respectively. Human liver microsomes exhibited a V(max) value of 26.9 nmol/min/mg protein and a K(m) value of 1790 microM. Hydrolytic activity of the erythrocytes and plasma was inhibited by 5, 5'-dithiobis(2-nitrobenzoic acid), an arylesterase inhibitor, in a concentration-dependent manner. In contrast, little or no suppression of these activities was seen with phenylmethylsulfonyl fluoride (PMSF), diisopropyl fluorophosphate (DFP), bis(p-nitrophenyl)phosphate (BNPP), BW284C51 and ethopropazine. The liver microsomal activity was markedly inhibited by PMSF, DFP and BNPP, indicating that carboxylesterase was involved in the nafamostat hydrolysis. Human carboxylesterase 2 expressed in COS-1 cells was capable of hydrolyzing nafamostat at 10 and 100 microM, whereas recombinant carboxylesterase 1 showed significant activity only at a higher substrate concentration (100 microM). The nafamostat hydrolysis in 18 human liver microsomes correlated with aspirin hydrolytic activity specific for carboxylesterase 2 (r=0.815, p<0.01) but not with imidapril hydrolysis catalyzed by carboxylesterase 1 (r=0.156, p=0.54). These results suggest that human arylesterases and carboxylesterase 2 may be predominantly responsible for the metabolism of nafamostat in the blood and liver, respectively. [Abstract/Link to Full Text]

Iwao Y, Anraku M, Hiraike M, Kawai K, Nakajou K, Kai T, Suenaga A, Otagiri M
The structural and pharmacokinetic properties of oxidized human serum albumin, advanced oxidation protein products (AOPP).
Drug Metab Pharmacokinet. 2006 Apr;21(2):140-6.
To determine the pharmacokinetic properties of advanced oxidation protein products (AOPP), we prepared oxidized human serum albumin (oxi-HSA) using chloramine-T (a hypochlorite analogue) in vitro. The AOPP and dityrosine content of oxi-HSA (AOPP content, 244.3+/-12.3 microM; dityrosine content, 0.7+/-0.11 nmol of dityrosine/mg protein) were similar to those of uremic patients. In structural analysis, the increases in AOPP and dityrosine content of HSA induced slight decreases in its alpha-helical content. In pharmacokinetic analysis, oxi-HSA left the circulation rapidly, and organ distribution of oxi-HSA 30 min after intravenous injection was 51% for the liver, 23% for the spleen, and 9% for the kidney, suggesting that the liver and spleen were the main routes of plasma clearance of oxi-HSA. The liver and spleen uptake clearance of oxi-HSA were significantly greater than those of normal HSA (CLliver, 5058+/-341.6 vs 24+/-4.2 microL/hr [p<0.01]; CLspleen, 2118+/-322.1 vs 32+/-2.7 microL/hr [p<0.01]). However, uptake by other organs was not significantly affected by oxidation. These results suggest that the liver and spleen play important roles in elimination of AOPP. [Abstract/Link to Full Text]

Nakade S, Ueda S, Ohno T, Nakayama K, Miyata Y, Yukawa E, Higuchi S
Population pharmacokinetics of pranlukast hydrate dry syrup in children with allergic rhinitis and bronchial asthma.
Drug Metab Pharmacokinet. 2006 Apr;21(2):133-9.
This study aimed to assess the steady-state pharmacokinetics of pranlukast, a leukotriene receptor antagonist, in children with allergic rhinitis and bronchial asthma, and to clarify factors affecting apparent clearance (CL/F). A total of 192 plasma samples were obtained from 98 children (rhinitis 64, asthma 13, complications 21), aged 3-14 years in 2 clinical trials. Plasma concentration of pranlukast was determined by liquid chromatography connected with a tandem mass spectrometer and analyzed by a population approach using NONMEM program. The plasma concentration-time course of pranlukast was described by using a one-compartment model with the first-order absorption and lag time. The robustness of the population pharmacokinetic model was evaluated by using 200 bootstrap samples. The results of population pharmacokinetic analysis showed that only age was a factor affecting the CL/F per body weight, with CL/F decreasing with increasing age. No significant variation was seen in the CL/F between rhinitis and asthma. The interindividual variability in the CL/F and the residual variability were 19.7% and 48.4%, respectively. All the parameters fell within 10% of the bootstrapped mean. In conclusion, the results show that age is the most influential factor for explaining interindividual variability in CL/F, and the difference in diseases does not affect CL/F. [Abstract/Link to Full Text]

Komoto C, Nakamura T, Sakaeda T, Kroetz DL, Yamada T, Omatsu H, Koyama T, Okamura N, Miki I, Tamura T, Aoyama N, Kasuga M, Okumura K
MDR1 haplotype frequencies in Japanese and Caucasian, and in Japanese patients with colorectal cancer and esophageal cancer.
Drug Metab Pharmacokinet. 2006 Apr;21(2):126-32.
The genotype frequencies of MDR1 T-129C, C1236T, G2677A,T and C3435T SNPs were compared in 154 healthy Japanese and 100 healthy Caucasians to provide basic information on the inter-ethnic differences of pharmacotherapeutic outcome. The variants were found at allelic frequencies of 5.5%, 65.6%, 16.6%, 40.6% and 40.6%, for T-129C, C1236T, G2677A, G2677T and C3435T, respectively, in Japanese, and at 5.1%, 45.9%, 3.6%, 46.4% and 56.6%, respectively, in Caucasians, with a statistically significant difference for C1236T, G2677A,T and C3435T (p<0.001). G2677A was about 5-fold more frequent in Japanese than Caucasians. These genotype frequencies were also investigated in 95 Japanese patients with colorectal cancer (CRC) and esophageal squamous cell carcinoma (ESCC), but no significant difference was detected, when compared with healthy Japanese subjects. The haplotype frequency reached a total of about 85% in Japanese with the following 4 major haplotypes; T(-129)-T1236-T2677-T3435 (36.1%), T(-129)-T1236-G2677-C3435 (22.5%), T(-129)-C1236-G2677-C3435 (14.2%) and T(-129)-C1236-A2677-C3435 (13.3%). The second and fourth haplotypes were hardly inferred in Caucasian, whereas T(-129)-C1236-G2677-T3435 (12.8%) was found to be Caucasian-specific. There was a tendency for higher frequencies of the T(-129)/C-(129)-C1236-A2677-C3435 haplotype in Japanese CRC patients and T(-129)-T1236-T2677-T3435 haplotype in Japanese ESCC patients, compared with that in healthy Japanese subjects. [Abstract/Link to Full Text]

Fukatsu S, Fukudo M, Masuda S, Yano I, Katsura T, Ogura Y, Oike F, Takada Y, Inui K
Delayed effect of grapefruit juice on pharmacokinetics and pharmacodynamics of tacrolimus in a living-donor liver transplant recipient.
Drug Metab Pharmacokinet. 2006 Apr;21(2):122-5.
Tacrolimus is a calcineurin inhibitor that has been widely used to prevent allograft rejection after transplantation. We report a case of a living-donor liver transplant recipient experiencing a considerable increase in the trough blood concentration of tacrolimus after concomitant ingestion of grapefruit juice (250 mL) 4 times for 3 days. The trough blood concentrations of tacrolimus were not changed during or immediate after the repeated intake of grapefruit juice. However, almost 1 week after the final ingestion, the blood concentration of tacrolimus markedly increased to as much as 47.4 ng/mL from 4.7 ng/mL before the ingestion, resulting in a profound reduction of calcineurin phosphatase activity in peripheral blood mononuclear cells. Furthermore, headache and nausea, but not nephrotoxicity or hyperglycemia, took place throughout the period of the elevated blood concentrations. Grapefruit juice may have a clinically significant effect on the pharmacokinetics and pharmacodynamics of tacrolimus. It is recommended to avoid the consumption of grapefruit juice in transplant recipients treated with tacrolimus. [Abstract/Link to Full Text]

Maekawa K, Itoda M, Sai K, Saito Y, Kaniwa N, Shirao K, Hamaguchi T, Kunitoh H, Yamamoto N, Tamura T, Minami H, Kubota K, Ohtsu A, Yoshida T, Saijo N, Kamatani N, Ozawa S, Sawada J
Genetic variation and haplotype structure of the ABC transporter gene ABCG2 in a Japanese population.
Drug Metab Pharmacokinet. 2006 Apr;21(2):109-21.
The ATP-binding cassette transporter, ABCG2, which is expressed at high levels in the intestine and liver, functions as an efflux transporter for many drugs, including clinically used anticancer agents such as topotecan and the active metabolite of irinotecan (SN-38). In this study, to elucidate the linkage disequilibrium (LD) profiles and haplotype structures of ABCG2, we have comprehensively searched for genetic variations in the putative promoter region, all the exons, and their flanking introns of ABCG2 from 177 Japanese cancer patients treated with irinotecan. Forty-three genetic variations, including 11 novel ones, were found: 5 in the 5'-flanking region, 13 in the coding exons, and 25 in the introns. In addition to 9 previously reported nonsynonymous single nucleotide polymorphisms (SNPs), 2 novel nonsynonymous SNPs, 38C>T (Ser13Leu) and 1060G>A (Gly354Arg), were found with minor allele frequencies of 0.3%. Based on the LD profiles between the SNPs and the estimated past recombination events, the region analyzed was divided into three blocks (Block -1, 1, and 2), each of which spans at least 0.2 kb, 46 kb, and 13 kb and contains 2, 24, and 17 variations, respectively. The two, eight, and five common haplotypes detected in 10 or more patients accounted for most (>90%) of the haplotypes inferred in Block -1, Block 1, and Block 2, respectively. The SNP and haplotype distributions in Japanese were different from those reported previously in Caucasians. This study provides fundamental information for the pharmacogenetic studies investigating the relationship between the genetic variations in ABCG2 and pharmacokinetic/pharmacodynamic parameters. [Abstract/Link to Full Text]

Noracharttiyapot W, Nagai Y, Matsubara T, Miyata M, Shimada M, Nagata K, Yamazoe Y
Construction of several human-derived stable cell lines displaying distinct profiles of CYP3A4 induction.
Drug Metab Pharmacokinet. 2006 Apr;21(2):99-108.
Cell lines which stably express reporter proteins through CYP3A4 gene activation have been developed for use in predicting CYP3A4 induction. Twelve clones showing distinct profiles on chemical-induced response were isolated. Among them, two clones showing high response for CYP3A4 inducers, namely clone 3-1-10 and 3-1-20, were further evaluated for their sensitivities, reproducibilities and applicabilities to predict CYP3A4 induction in human. Clone 3-1-10 showed higher response to rifampicin than to clotrimazole, whereas clone 3-1-20 had rather higher response to clotrimazole. Optimal plating density and highly reproducible response were observed at the range of 1.65-5.0 x 10(4) cell/cm2. Clear induction responses of more than ten chemicals were observed in both cell lines. The reporter activity was further dramatically increased after an introduction of human PXR. Induction with rifampicin was, however, not much altered between the absence and presence of hPXR. The luciferase activity remained unaltered and showed little fluctuation during the culture for more than 6 months. Due to the strikingly high sensitivity and reproducibility of this system, as compared to previously published systems, these HepG2-derived cell lines showing distinct response profiles as developed in the present study will offer high advantages for chemical screening of CYP3A4 inducibility. [Abstract/Link to Full Text]

Kitamura S, Sugihara K, Ohta S
Drug-metabolizing ability of molybdenum hydroxylases.
Drug Metab Pharmacokinet. 2006 Apr;21(2):83-98.
Molybdenum hydroxylases, which include aldehyde oxidase and xanthine oxidoreductase, are involved in the metabolism of some medicines in humans. They exhibit oxidase activity towards various heterocyclic compounds and aldehydes. The liver cytosol of various mammals also exhibits a significant reductase activity toward nitro, sulfoxide, N-oxide and other moieties, catalyzed by aldehyde oxidase. There is considerable variability of aldehyde oxidase activity in liver cytosol of mammals: humans show the highest activity, rats and mice show low activity, and dogs have no detectable activity. On the other hand, xanthine oxidoreductase activity is present widely among species. Interindividual variation of aldehyde oxidase activity is present in humans. Drug-drug interactions associated with aldehyde oxidase and xanthine oxidoreductase are of potential clinical significance. Drug metabolizing ability of molybdenum hydroxylases and the variation of the activity are described in this review. [Abstract/Link to Full Text]

Fujita K, Ando Y, Nagashima F, Yamamoto W, Endo H, Kodama K, Araki K, Miya T, Narabayashi M, Sasaki Y
Novel single nucleotide polymorphism of UGT1A9 gene in Japanese.
Drug Metab Pharmacokinet. 2006 Feb;21(1):79-81.
We sequenced from 5'-franking region to intron 1 (to 337 bp downstream from exon 1) of the UDP-glucuronosyltransferase (UGT) 1A9 gene prepared from 55 Japanese cancer patients. Seven single nucleotide polymorphisms (SNPs) were found. Two of them were UGT1A9 -118(T)n (n=10) and UGT1A9*5, and four were reported SNPs in intron 1 of UGT1A9 gene (89540C>T, 89549G>A, 89616T>A and 89710A>C). A novel SNP (89587T>C) was found. The sequence is as follows: SNP, 050824FujitaK001; Gene Name, UGT1A9; Accession Number, AF297093; Length, 25 bases; 5'-CCTTCTTGAAGAT/CATGTATTTATAA-3'. Two patients were heterozygous for the mutant allele, resulting in the allele frequency of 1.82%. [Abstract/Link to Full Text]

Fujita K, Ando Y, Nagashima F, Yamamoto W, Endo H, Kodama K, Araki K, Miya T, Narabayashi M, Sasaki Y
Novel single nucleotide polymorphism of UGT1A7 gene in Japanese.
Drug Metab Pharmacokinet. 2006 Feb;21(1):75-8.
We sequenced exon 1 of the UDP-glucuronosyltransferase (UGT) 1A7 gene from 52 Japanese cancer patients. Four single nucleotide polymorphisms (SNPs) were found. Three of them caused UGT1A7*2 and UGT1A7*3. A novel SNP (98973G>C) causing amino acid substitution (Ser141Cys) was found. The sequence is as follows: SNP, 050824FujitaK002; Gene Name, UGT1A7; Accession Number, AF297093; Length, 25 bases; 5'-TAAAGGAGAGTTG/CTTTTGATGCAGT-3'. One out of 52 cancer patients was heterozygous for the variant allele, resulting in the allele frequency of 0.96%. The patient did not possess UGT1A7*2 or UGT1A7*3. [Abstract/Link to Full Text]

Ikushiro S, Emi Y, Kato Y, Yamada S, Sakaki T
Monospecific antipeptide antibodies against human hepatic UDP-glucuronosyltransferase 1A subfamily (UGT1A) isoforms.
Drug Metab Pharmacokinet. 2006 Feb;21(1):70-4.
Expression of UDP-glucuronosyltransferases (UGT) in mammals is thought to be regulated in both a tissue- and developmental-specific manner. Furthermore, induction of genes encoding UGT occurs after exposure to xenobiotics including drugs, environmental pollutants and dietary compounds. In human, isoforms of UGT 1A subfamily catalyze the glucuronidation of a greater proportion of drugs, suggesting that the expression of UGT1A isoforms is responsible for the clearance of a diverse range of drugs. To analyze the expression of human UGT1A isoforms, we have developed polyclonal antibodies against specific peptide regions within the isoforms (UGT1A1, 1A3, 1A4, 1A6 and 1A9). The prepared antipeptide antibodies were found to be highly monospecific for each UGT1A isoform and no cross-reactivity with UGT2B isoforms was detected. Analysis of UGT1A protein levels in hepatic microsomes using these antibodies demonstrated interindividual differential expression of each isoform. These highly specific antipeptide antibodies provide an important tool to analyze tissue distribution and interindividual expression levels of human UGT1As. [Abstract/Link to Full Text]

Tanigawa T, Heinig R, Kuroki Y, Higuchi S
Evaluation of interethnic differences in repinotan pharmacokinetics by using population approach.
Drug Metab Pharmacokinet. 2006 Feb;21(1):61-9.
Repinotan is a selective full serotonin receptor agonist at the 5-HT1A subtype which has been studied in phase I and II studies involving over 500 healthy subjects and patients. Repinotan is primarily metabolized by CYP2D6 which is known to be subject to polymorphism and ethnic differences in its quantitative and qualitative expression pattern. In order to investigate the effect of ethnicity on repinotan pharmacokinetics (PK) between a Caucasian and Japanese population and to explain PK variability, this population PK evaluation was conducted. A population PK model was established based on the data of 1314 blood samples from 241 patients from 3 Phase II studies. This analysis has characterized the repinotan PK, with particular attention to ethnicity. Using the MIXTURE subroutine of NONMEM, evidence was provided for different CL groups. Repinotan plasma levels in the 'High CL' subgroup, which comprised the majority of patients, did not show relevant differences between a Japanese and Caucasian population. In the 'Low CL' subgroup, Japanese and Caucasian populations were different. These findings are consistent with the published literature, which reports ethnic differences in the distribution of CYP2D6 activity. The finding of a greater percentage of patients with intermediate CL in the Japanese population falling between poor and extensive metabolizers is consistent with the distribution pattern of CYP2D6 in the Japanese population. The results of this evaluation can be used to assist in designing future trials. [Abstract/Link to Full Text]

Maeda Y, Omoda K, Fukuhara S, Ohta M, Ishii Y, Murakami T
Evaluation of clinical efficacy of Maeda's nomogram for vancomycin dosage adjustment in adult Japanese MRSA pneumonia patients.
Drug Metab Pharmacokinet. 2006 Feb;21(1):54-60.
The clinical efficacy of Maeda's nomogram for vancomycin dosage adjustment was evaluated by comparison with a standard dosage regimen (package insert information: vancomycin dose reduced in elderly patients and patients with renal dysfunction, with Moellering's nomogram used for renal-dysfunction patients) in adult Japanese MRSA pneumonia patients. Using Maeda's nomogram, the vancomycin dose is fixed at 1,000 mg while the dosing interval is varied in accordance with individual creatinine clearance. Using a standard dosage regimen, 5 patients out of 27 (18.5%) achieved target plasma levels of vancomycin (25-40 microg/mL for peak and 5-15 microg/mL for trough) within 2-6 days. Using Maeda's nomogram, 38 patients out of 53 (71.7%) achieved target levels in that time. A higher clinical response (complete resolution of all signs and symptoms of MRSA infection) to vancomycin therapy was also obtained with Maeda's nomogram when evaluated approximately 2-weeks after discontinuation of vancomycin therapy (43.4% versus 18.5% for the standard regimen). In conclusion, the Maeda's nomogram regimen with a 1,000 mg vancomycin dose was shown to achieve target plasma levels of vancomycin at a higher rate and provide higher clinical efficacy in vancomycin therapy of MRSA pneumonia patients, as compared with the currently available standard dosage regimen. [Abstract/Link to Full Text]

Araya H, Tomita M, Hayashi M
The novel formulation design of self-emulsifying drug delivery systems (SEDDS) type O/W microemulsion III: the permeation mechanism of a poorly water soluble drug entrapped O/W microemulsion in rat isolated intestinal membrane by the Ussing chamber method.
Drug Metab Pharmacokinet. 2006 Feb;21(1):45-53.
We used ibuprofen as a poorly water soluble model drug, to examine the influence of bile salts and mucin layers on the permeability of that entrapped in an O/W microemulsion, in a rat isolated intestinal membrane by the Ussing chamber method. Under the presence of 3 kinds of the primary bile salts such a sodium taurocholate, etc., or a secondary bile salt such a sodium taurochenodeoxycholate at 0.01 mmol/L concentration, a significant difference was not demonstrated in the permeation clearance of the ibuprofen entrapped O/W microemulsion, as compared with the case without the bile salts. Thus, the bile salts did not have a remarkable influence on the permeability of the drug entrapped in the O/W microemulsion, and it was verified that this O/W microemulsion was hardly influenced by the flow of the bile secretion. On the other hand, when N-acetyl-L-cysteine (NAC) with the removal ability of a mucin layer was combined with the ibuprofen entrapped O/W microemulsion at the concentration of 3 and 10 mmol/L, it was shown that the permeation clearance of free ibuprofen did not decrease, but that of ibuprofen entrapped in the O/W microemulsion decreased with the increase of the NAC concentration. Therefore, it is confirmed that the mucin layer participates in the permeability of the drug entrapped in the O/W microemulsion. From these results, the mechanism in which the drug entrapped in the O/W microemulsion is released in a mucin layer, without passing through the route of the mixed micelle formation by bile, thereafter the drug permeates an intestinal membrane, is supposed. [Abstract/Link to Full Text]

Chono S, Tauchi Y, Morimoto K
Pharmacokinetic analysis of the uptake of liposomes by macrophages and foam cells in vitro and their distribution to atherosclerotic lesions in mice.
Drug Metab Pharmacokinet. 2006 Feb;21(1):37-44.
In order to evaluate the efficacy of liposomes as a drug carrier for atherosclerotic therapy, a pharmacokinetic analysis of the uptake of liposomes by macrophages and foam cells in vitro and their distribution to atherosclerotic lesions in mice was carried out. In brief, liposomes of three particle sizes (500, 200 and 70 nm) were prepared, and the uptake of liposomes by these cells in vitro and the aortic distribution following intravenous administration to atherogenic mice were examined. The internalization rate constant calculated by measuring uptake and binding was size-dependent in both types of cells in vitro. The aortic clearance (CL(a)) was size-independent in atherogenic mice and the CL(a) of 200 nm particles was the highest. Surprisingly, the aortic distribution in vivo did not correspond with the internalization to macrophages and foam cells in vitro. These results suggest that there is an optimal size for the distribution of liposomes to atherosclerotic lesions. [Abstract/Link to Full Text]

Jarukamjorn K, Sakuma T, Jaruchotikamol A, Oguro M, Nemoto N
Regulation of mouse hepatic CYP2D9 mRNA expression by growth and adrenal hormones.
Drug Metab Pharmacokinet. 2006 Feb;21(1):29-36.
The constitutive expression of CYP2D9 is sexually dimorphic, namely, strong in males, but diminutive in females. Repetition of mimic growth hormone (GH) secretion pattern impressively returned the mRNA expression level to that in intact mice: the GH secretion pattern's regulation of CYP2D9 mRNA expression has been predominantly disrupted by exogenous GH-administration. The extensive decline of CYP2D9 mRNA expression becoming a sexually non-specific P450 in 9-week-old male mice exposed as neonates to monosodium L-glutamate (MSG) suggested that the male GH secretion pattern is a key to the regulation of male-specific CYP2D9 mRNA expression in adult mice. Dexamethasone (Dex) showed possibility to induce CYP2D9 mRNA expression in adult MSG-neonatally treated mice of either sex. However, the antagonism was observed by co-administration of Dex and GH in the males. Dex-administration in adrenalectomized mice significantly elevated CYP2D9 mRNA expression levels. These findings suggest that an adrenal hormone participates in the regulatory mechanism of CYP2D9 mRNA expression in association with GH. [Abstract/Link to Full Text]

Morioka Y, Nishimura M, Imai T, Suzuki S, Harada M, Satoh T, Naito S
Assessment of induction of cytochrome P450 by NO-1886 (ibrolipim), a lipoprotein lipase-promoting agent, in primary cultures of human hepatocytes and in female rat liver.
Drug Metab Pharmacokinet. 2006 Feb;21(1):19-28.
The mRNA levels of human cytochrome P450 (CYP)2Cs and CYP3As in primary cultures of freshly isolated human hepatocytes were assessed after exposure to NO-1886 and rifampicin, a typical inducer of CYP3As. mRNA levels were analyzed by real-time RT-PCR using an ABI PRISM 7700 Sequence Detector system. Exposure to NO-1886 for 24 hr at a concentration of less than 10 microM showed only a tendency to reduce or increase the expression levels of CYP2C8, CYP2C9, CYP2C19, CYP3A4, or CYP3A5 mRNA. A higher concentration (50 microM) of NO-1886 induced an increase in CYP2C8 mRNA and a decrease in CYP2C19 mRNA, and these changes continued after additional culture for 24 hr in fresh medium without NO-1886. The expression level of CYP3A4 mRNA after exposure to NO-1886 for 24 hr at 50 microM was about twice that in controls. Following additional culture for 24 hr in fresh medium without NO-1886, the expression of CYP3A4 mRNA was comparable to that in controls. On the other hand, the expression levels of CYP2C9 and CYP3A5 mRNA showed small and variable changes in each donor even at a high concentration (50 microM) of NO-1886. Furthermore, the pharmacokinetics of NO-1886 during repeated oral administration for 14 days was studied in female rats. The pharmacokinetic parameters of NO-1886 were nearly the same on days 1, 7, and 14 of repeated administration. The hepatic microsomal content of CYP isoforms was not affected by repeated administration for 7 days at a dose of 1 to 30 mg/kg in female rats, although the total CYP content was increased at a dose of 30 mg/kg. The expression levels of CYP1A2, CYP2B2, CYP2C12, and CYP2E1 mRNA in primary cultures of rat hepatocytes were not affected by exposure to NO-1886 at 2, 10, or 50 microM. The expression levels of CYP3A1 mRNA in primary cultures of rat hepatocytes were not affected by exposure to NO-1886 at 2 or 10 microM, but were increased, with large individual variation, by exposure at 50 microM. The mRNA expression levels in rat hepatocytes exposed to concentrations comparable to free plasma levels did not change significantly, which was consistent with the equivalence in the in vivo plasma concentrations observed on days 1 and 14 of repeated administration. These results suggest that repeated administration of NO-1886 at clinical doses does not significantly affect the expression levels of CYP isoforms in human liver, although the mRNA levels of the CYP isoforms involved in the metabolism of NO-1886 were increased by exposure to higher concentrations of NO-1886 in human hepatocytes in vitro. [Abstract/Link to Full Text]

Matsunaga T, Shintani S, Hara A
Multiplicity of mammalian reductases for xenobiotic carbonyl compounds.
Drug Metab Pharmacokinet. 2006 Feb;21(1):1-18.
A variety of carbonyl compounds are present in foods, environmental pollutants, and drugs. These xenobiotic carbonyl compounds are metabolized into the corresponding alcohols by many mammalian NAD(P)H-dependent reductases, which belong to the short-chain dehydrogenase/reductase (SDR) and aldo-keto reductase superfamilies. Recent genomic analysis, cDNA isolation and characterization of the recombinant enzymes suggested that, in humans, the six members of each of the two superfamilies, i.e., total of 12 enzymes, are involved in the reductive metabolism of xenobiotic carbonyl compounds. They comprise three types of carbonyl reductase, dehydrogenase/reductase (SDR family) member 4, 11beta-hydroxysteroid dehydrogenase type 1, L-xylulose reductase, two types of aflatoxin B1 aldehyde reductase, 20alpha-hydroxysteroid dehydrogenase, and three types of 3alpha-hydroxysteroid dehydrogenase. Accumulating data on the human enzymes provide new insights into their roles in cellular and molecular reactions including xenobiotic metabolism. On the other hand, mice and rats lack the gene for a protein corresponding to human 3alpha-hydroxysteroid dehydrogenase type 3, but instead possess additional five or six genes encoding proteins that are structurally related to human hydroxysteroid dehydrogenases. Characterization of the additional enzymes suggested their involvement in species-specific biological events and species differences in the metabolism of xenobiotic carbonyl compounds. [Abstract/Link to Full Text]

Sasabe H, Kato Y, Suzuki T, Itose M, Miyamoto G, Sugiyama Y
Carrier-mediated uptake of grepafloxacin, a fluoroquinolone antibiotic, by the isolated rat lung cells.
Drug Metab Pharmacokinet. 2005 Dec;20(6):491-5.
Grepafloxacin (GPFX) is a new quinolone antibiotic (NQ) which is highly distributed to the lung and other tissues. In the present study, to characterize the distribution mechanism of GPFX to the lung, the uptake of GPFX by isolated rat lung cells was examined in vitro. GPFX was rapidly taken up by the cells, and the uptake reached a steady-state within 5 min. The cell-to-medium concentration ratio at equilibrium was 56.8+/-1.9 microL/mg protein, which was much higher than the cellular volume. GPFX uptake consisted of a saturable component (Km: 264+/-181 microM, Vmax: 2.94+/-2.33 nmol/min/mg protein) and a nonsaturable component (Pdif: 7.04+/-2.17 microL/min/mg protein). The uptake of GPFX was reduced in the presence of ATP-depletors (FCCP and Rotenone) and by the replacement of sodium with choline in the medium, suggesting that GPFX uptake is at least partially mediated by an Na+- and energy-dependent process. GPFX uptake tended to be reduced in the presence of other NQs such as levofloxacin, lomefloxacin and sparfloxacin, but was only minimally affected by the substrates of several uptake mechanisms already identified in the liver and kidney such as taurocholate, p-aminohippurate, L-carnitine and tetraethylammonium. These results suggested that GPFX is taken up by the lung partially via carrier-mediated transport system(s), distinct from the identified transporters, and such active transport systems may at least partially account for the efficient distribution of GPFX to the lung. [Abstract/Link to Full Text]

Aoyama T, Ogata K, Shimizu M, Hatta S, Masuhara K, Shima Y, Kimura K, Matsumoto Y
Pharmacokinetics of fluconazole and fosfluconazole after intraperitoneal administration to peritoneal dialysis rats.
Drug Metab Pharmacokinet. 2005 Dec;20(6):485-90.
Fluconazole (FLCZ) is an antifungal agent that is efficacious in the treatment of fungal peritonitis. Fosfluconazole (F-FLCZ) is the phosphate prodrug of FLCZ, which is highly soluble compared with FLCZ. F-FLCZ is useful against fungal peritonitis in continuous ambulatory peritoneal dialysis (CAPD) patients because it has a high water solubility. The aims of the present study were to characterize the peritoneal permeability of FLCZ and the pharmacokinetics of FLCZ and F-FLCZ after intraperitoneal (i.p.) administration to peritoneal dialysis rats. FLCZ or F-FLCZ was administered intravenously and intraperitoneally. After the i.p. administration of F-FLCZ, FLCZ was detected in circulating blood and the dialyzing fluid in peritoneal dialysis rats. The concentration of plasma FLCZ after the i.p. F-FLCZ administration was lower than that after the intravenous (i.v.) F-FLCZ administration. It is considered that the dose should be increased appropriately when F-FLCZ is administered intraperitoneally. The profiles of plasma FLCZ after i.v. and i.p. administrations were analyzed using a two-compartment model in which the distribution volume of the peripheral compartment was fixed at a volume of the dialyzing fluid (peritoneal dialysis PK model). The peritoneal dialysis PK model could describe the profiles of plasma and dialyzing fluid FLCZ. These results suggest that FLCZ and F-FLCZ could be administered intraperitoneally for the treatment of fungal peritonitis in CAPD patients. [Abstract/Link to Full Text]

Imaoka S, Hashizume T, Funae Y
Localization of rat cytochrome P450 in various tissues and comparison of arachidonic acid metabolism by rat P450 with that by human P450 orthologs.
Drug Metab Pharmacokinet. 2005 Dec;20(6):478-84.
Metabolites of arachidonic acid produced by P450 are interesting substances with prominent physiological functions. To elucidate the physiological function of P450, it is necessary to identify a specific P450 in a particular tissue or organ and to characterize its catalytic activities. In this study, the expression of CYP2A1, 2B1, 2C23, 2J3, and 4F1 was investigated in liver, lung, kidney, spleen, heart, brain, and testis of rats by RT-PCR. Furthermore, arachidonic acid metabolism was investigated using the rat P450s described above and human CYP2A6, 2B6, 2C9, 2C18, 2C19, 2J2, and 4F2. Among the rat P450s, CYP2B1 and 2C23 efficiently produced EETs and CYP4F1 produced 19/20-HETE in abundace. CYP2B1 was specifically expressed in the lung. CYP2C23 was detected in all tissues used in this study. CYP4F1 was expressed in the kidney as well as in the liver. Among the human P450s, CYP2C9 and 2C19 efficiently produced EETs. CYP4F2 produced 19/20-HETE. The catalytic properties of rat CYP2C23 were similar to those of human CYP2C9 and 2C19. The catalytic properties of CYP4F isoforms were also similar between humans and rats. A systematic analysis of P450 expression in various tissues and of its catalytic property may provide valuable information on the physiological roles of P450s in each tissue. [Abstract/Link to Full Text]

Nishimura M, Naito S
Tissue-specific mRNA expression profiles of human ATP-binding cassette and solute carrier transporter superfamilies.
Drug Metab Pharmacokinet. 2005 Dec;20(6):452-77.
Pairs of forward and reverse primers and TaqMan probes specific to each of 46 human ATP-binding cassette (ABC) transporters and 108 human solute carrier (SLC) transporters were prepared. The mRNA expression level of each target transporter was analyzed in total RNA from single and pooled specimens of various human tissues (adrenal gland, bone marrow, brain, colon, heart, kidney, liver, lung, pancreas, peripheral leukocytes, placenta, prostate, salivary gland, skeletal muscle, small intestine, spinal cord, spleen, stomach, testis, thymus, thyroid gland, trachea, and uterus) by real-time reverse transcription PCR using an ABI PRISM 7700 sequence detector system. In contrast to previous methods for analyzing the mRNA expression of single ABC and SLC genes such as Northern blotting, our method allowed us to perform sensitive, semiautomatic, rapid, and complete analysis of ABC and SLC transporters in total RNA samples. Our newly determined expression profiles were then used to study the gene expression in 23 different human tissues, and tissues with high transcriptional activity for human ABC and SLC transporters were identified. These results are expected to be valuable for establishing drug transport-mediated screening systems for new chemical entities in new drug development and for research concerning the clinical diagnosis of disease. [Abstract/Link to Full Text]

Watanabe C, Kato Y, Ito S, Kubo Y, Sai Y, Tsuji A
Na+/H+ exchanger 3 affects transport property of H+/oligopeptide transporter 1.
Drug Metab Pharmacokinet. 2005 Dec;20(6):443-51.
Oligopeptide transporter PEPT1 is thought to be involved in the intestinal absorption and renal reabsorption of peptides and therapeutic agents. The driving force of PEPT1 is H+ gradient, a part of which is supplied by Na+/H+ exchanger (NHE) expressed on the apical surface of the epithelium although molecular identification of NHE has not yet been fully clarified. Here we examined the effect of NHE3 coexpression on the function of PEPT1 to support the hypothesis that NHE3 regulates PEPT1 function by supplying its driving force. HEK293 cells expressing PEPT1 alone exhibited Na+-independent but pH-dependent uptake of glycylsarcosine (GlySar), whereas those coexpress PEPT1 and NHE3 showed an increase in GlySar uptake and conferred Na+-dependence on the uptake of GlySar. The increase in GlySar transport by PEPT1 depended on the expression level of NHE3 and was found at various levels of PEPT1 expression. Kinetic analysis of GlySar uptake in HEK293 cells expressing both PEPT1 and NHE3 or those expressing PEPT1 alone revealed an approximately 3 times increase in the transport capacity in the presence of NHE3, as normalized by PEPT1 mRNA expression. Confocal microscopy indicated that both PEPT1 and NHE3 are colocalized on the cell-surface of HEK293 cells. Thus, the present findings are the first to specify that NHE3 exerts post-transcriptional stimulation of PEPT1-mediated transport and can affect cellular uptake of the substrates by PEPT1 expressed on apical membranes in the body. [Abstract/Link to Full Text]

Kaneo Y, Hashihama S, Kakinoki A, Tanaka T, Nakano T, Ikeda Y
Pharmacokinetics and biodisposition of poly(vinyl alcohol) in rats and mice.
Drug Metab Pharmacokinet. 2005 Dec;20(6):435-42.
Poly(vinyl alcohol) (PVA) of various molecular weight (MW=10,560-116,600) was successfully labeled with fluorescein isothiocyanate isomer I (FITC) according to the method of de Belder and Granath. A high-performance size-exclusion chromatographic procedure was developed for the quantitative analysis of FITC-labeled poly(vinyl alcohol) (F-PVA) in biological samples. F-PVA (80 K) disappeared slowly from the blood circulation according to the first-order kinetics (t1/2=7 h) after intravenous injection to rats. A dose-independent behavior of F-PVA (80 K) was observed in the blood circulation, in the tissue distribution and in the urinary and fecal excretions. This suggested that PVAs are eliminated exclusively by the mechanisms that do not involve saturable transport processes. Furthermore, it was found that PVAs are very stable in the body because no degradation product was detected in the urine and feces. 125I-labeled poly(vinyl alcohol) (125I-PVA) was prepared by introducing tyramine residues to the hydroxyl groups of PVA molecules by the 1,1'-cabonyldiimidazole (CDI) activation method. 125I-PVA (80 K) was retained in the blood circulation for several days after intravenous injection to mice. Although the tissue distribution of PVAs was small, a significant accumulation into the liver and the spleen was observed. Fluorescence microscopic examination of paraffin section of the liver revealed that F-PVA (80 K) was endocytosed by the liver parenchymal cells. 125I-PVA (80 K) captured by liver was slowly transported via the bile canaliculi and gall bladder to the intestine and excreted in the feces. It was suggested, therefore, a long time is necessary for 125I-PVA (80 K) to be excreted perfectly from the body. [Abstract/Link to Full Text]

Watanabe E, Kinoshita N, Takahashi M, Hayashi M
Prediction of absorbability of poorly water-soluble drugs based on permeability obtained through modified in vitro chamber method.
Drug Metab Pharmacokinet. 2005 Dec;20(6):428-34.
Permeability is an underlying parameter to control drug absorption. For highly water-soluble drugs, the high correlation between their permeability and fraction absorbed in humans is reported. In the present study, to predict the absorbability of poorly water-soluble drugs in humans, a new experimental method of the permeation study was proposed and subjected to examination. Firstly, using the in vitro chamber method modified to contain 5% (final concentration) dimethyl sulufoxide (DMSO) in both compartments of the chamber (DMSO-MS), the effect of DMSO on membrane integrity was evaluated. Secondly, the correlation between the apparent permeability coefficients (Papp) obtained through DMSO-M or DMSO-MS and fractions absorbed in humans were investigated using 7 poorly water-soluble drugs. Membrane integrity of the rat intestinal tissues was maintained after using DMSO-MS, as with that after using the conventional in vitro chamber method. Papp of two paracellular markers obtained through DMSO-MS was not different from that obtained through the conventional chamber method. In the permeation study of the P-glycoprotein substrate, Papp from both mucosal to serosal and serosal to mucosal sides obtained through DMSO-MS was not significantly different from that obtained through the conventional chamber method. The correlation between Papp obtained through DMSO-MS and Fa which was expressed by the equation of Fa=1-exp (-Pappx1.38x10(5)) (r2=0.980), was more favorable than the correlation between Papp obtained through DMSO-M and Fa which was expressed by the equation of Fa=1-exp (-Pappx2.12x10(5)) (r2=0.875). These results showed that DMSO-MS was a useful method for predicting the absorbability of poorly water-soluble drugs. [Abstract/Link to Full Text]

Fukumoto K, Kobayashi T, Komamura K, Kamakura S, Kitakaze M, Ueno K
Stereoselective effect of amiodarone on the pharmacokinetics of racemic carvedilol.
Drug Metab Pharmacokinet. 2005 Dec;20(6):423-7.
We investigated whether there was a stereoselective effect of amiodarone on the pharmacokinetics of carvedilol. Among a series of 106 inpatients with heart failure, 52 received carvedilol monotherapy (carvedilol group) and 54 received carvedilol plus amiodarone (carvedilol+amiodarone group). The serum carvedilol concentration administered/dose ratio was compared between the two groups based on HPLC measurement of the serum levels of carvedilol, amiodarone, and desethylamiodarone. In 6 patients from the carvedilol group, serum carvedilol levels were compared before and after coadministration of amiodarone. There was no significant between-group difference of the serum concentration to dose (C/D ratio) for the R-enantiomer carvedilol, however, the C/D ratio for the S-enantiomer and the serum S-carvedilol to R-carvedilol (S/R) ratio were both significantly lower in the carvedilol group than in the carvedilol+amiodarone group(47.8+/-56.7 versus 95.3+/-105 ng/mg/kg, P=0.0048 and 0.460+/-0.207 versus 0.879+/-0.377 ng/mg/kg, P<0.001), respectively. Furthermore, the mean S-carvedilol concentration over 14 days of coadministration with amiodarone was higher than that before coadministration (6.54+/-1.73 ng/mL versus 3.03+/-0.670 ng/mL, P<0.001). These results suggest that metabolism of S-carvedilol was markedly inhibited by coadministration of amiodarone. [Abstract/Link to Full Text]

Ohnishi A, Yano Y, Ishibashi T, Katsube T, Oguma T
Evaluation of Bayesian predictability of vancomycin concentration using population pharmacokinetic parameters in pediatric patients.
Drug Metab Pharmacokinet. 2005 Dec;20(6):415-22.
The objective of this study was to evaluate the Bayesian predictability of vancomycin (VCM) pharmacokinetics in Japanese pediatric patients using one-compartment population pharmacokinetic (PPK) parameters, which we reported previously. The validity of the PPK model was evaluated by bootstrap method and cross validation method, and the Bayesian predictive performance was examined. The predictive performance of the PPK model for premature patients was also examined. The cross validation method showed the predictability to be acceptable for practical use, especially for predicting trough concentration using other trough data. However, for the external premature patient data, this PPK model did not seem to be adequate. A theoretical approach using a simulation technique was also examined to evaluate the predictive performance. The results suggested that the predictability at the peak was not necessarily good at all sampling times and the predictability at the trough was better when a later time point was used. The optimal sampling time for prediction of VCM concentration in pediatric patients is discussed. [Abstract/Link to Full Text]

Recent Articles in Journal of Pharmacological Sciences

Lee CW, Lee SH, Lee JW, Ban JO, Lee SY, Yoo HS, Jung JK, Moon DC, Oh KW, Hong JT
2-hydroxycinnamaldehyde inhibits SW620 colon cancer cell growth through AP-1 inactivation.
J Pharmacol Sci. 2007 May;104(1):19-28.
Cinnamaldehyde derivatives isolated from Cinnamomum cassia have been widely used for treating dyspepsia, gastritis, and inflammatory disease as well as cancer. To investigate the anti-tumor activities of several cinnamaldehyde derivatives, we compared the inhibitory effect of cinnamaldehyde derivatives on cell growth and AP-1 transcriptional activity in SW620 human colon cancer cells since AP-1 is a transcriptional factor implicated to control cancer cell growth. Among the derivatives, 2'-hydroxycinnamaldehyde (HCA) most significantly inhibited cancer cell growth and AP-1 transcriptional activity in a dose-dependent manner with an IC50 value of 12.5 and 9 microg/ml, respectively. In further studies on the mechanism, we found that consistent with the inhibitory effect on cell growth, HCA dose-dependently (0-20 microg/ml) inhibited DNA binding activity of AP-1 accompanied with down regulation of c-Jun and c-Fos expressions. HCA also induced apoptotic cell death as well as expression of the apoptosis-regulating gene caspase-3, but inhibited the anti-apoptosis regulating gene bcl-2 in a dose-dependent manner. These results suggested that HCA has the most potent inhibitory effect against human colon cancer cell growth, and AP-1 may be an important target of HCA. [Abstract/Link to Full Text]

Yamazaki K, Inoue T, Yasuda N, Sato Y, Nagakura T, Takenaka O, Clark R, Saeki T, Tanaka I
Comparison of efficacies of a dipeptidyl peptidase IV inhibitor and alpha-glucosidase inhibitors in oral carbohydrate and meal tolerance tests and the effects of their combination in mice.
J Pharmacol Sci. 2007 May;104(1):29-38.
E3024 (3-but-2-ynyl-5-methyl-2-piperazin-1-yl-3,5-dihydro-4H-imidazo[4,5-d]pyridazin-4-one tosylate) is a dipeptidyl peptidase IV (DPP-IV) inhibitor. Since the target of both DPP-IV inhibitors and alpha-glucosidase inhibitors is the lowering of postprandial hyperglycemia, we compared antihyperglycemic effects for E3024 and alpha-glucosidase inhibitors in various oral carbohydrate and meal tolerance tests using normal mice. In addition, we investigated the combination effects of E3024 and voglibose on blood glucose levels in a meal tolerance test using mice fed a high-fat diet. ER-235516-15 (the trifluoroacetate salt form of E3024, 1 mg/kg) lowered glucose excursions consistently, regardless of the kind of carbohydrate loaded. However, the efficacy of acarbose (10 mg/kg) and of voglibose (0.1 mg/kg) varied with the type of carbohydrate administered. The combination of E3024 (3 mg/kg) and voglibose (0.3 mg/kg) improved glucose tolerance additively, with the highest plasma active glucagon-like peptide-1 levels. This study shows that compared to alpha-glucosidase inhibitors, DPP-IV inhibitors may have more consistent efficacy to reduce postprandial hyperglycemia, independent of the types of carbohydrate contained in a meal, and that the combination of a DPP-IV inhibitor and an alpha-glucosidase inhibitor is expected to be a promising option for lowering postprandial hyperglycemia. [Abstract/Link to Full Text]

Nishida H, Kushida M, Nakajima Y, Ogawa Y, Tatewaki N, Sato S, Konishi T
Amyloid-beta-induced cytotoxicity of PC-12 cell was attenuated by Shengmai-san through redox regulation and outgrowth induction.
J Pharmacol Sci. 2007 May;104(1):73-81.
Neurodegenerative brain disorders such as Alzheimer's disease (AD) have been well investigated. However, significant methods for the treatment of the promotion and progression of AD are unavailable to date. Recent studies suggested that the redox imbalance and the accumulation of amyloid-beta (Abeta) peptide occurring in the brain of AD patients lead to oxidatively-induced apoptotic cell death. Here, we show the effects of Shengmai-san (SMS) on Abeta-induced cytotoxicity in PC-12 cells. SMS dose-dependently attenuated the cytotoxicity by Abeta incubation and also prevented the morphological damage in neurites of the PC-12 cells. Hemeoxygenase-1 and glutathione peroxidase-1 expressions were increased by SMS pretreatment. SMS decreased the phosphorylation level of c-jun amino-terminal kinase (JNK) and the activity of caspase-3, which were enhanced by Abeta incubation. Of importance, SMS treatment promoted neurite outgrowth. These data demonstrated dual roles of SMS in PC-12 cells. SMS prevents the apoptosis through the enhancement of anti-oxidant enzymes and inhibition of the JNK signaling pathway with the promotion of nerve cell maturation, thus suggesting benefits of SMS for the treating of neurodegenerative diseases. It may also be beneficial not only for the treatment of brain disorders but also for other diseases caused by oxidative stress. [Abstract/Link to Full Text]

Yamazaki D, Ohya S, Asai K, Imaizumi Y
Characteristics of the ATP-induced Ca2+-entry pathway in the t-BBEC 117 cell line derived from bovine brain endothelial cells.
J Pharmacol Sci. 2007 May;104(1):103-7.
ATP-receptor (P2Y) stimulation induced sustained Ca2+-entry, which was essential for the enhanced cell-proliferation in t-BBEC117, an immortalized cell-line derived from bovine brain endothelial cells. Application of Ca2+ following store-depletion with thapsigargin in Ca2+-free solution induced Ca2+-entry through store-operated channels (SOCs). Ca2+-entry induced by ATP or 1-oleoyl-2-acetyl-sn-glycerol (OAG) together with Ca2+ was significantly larger than that by Ca2+ alone, suggesting the involvement of receptor-operated channels (ROCs) in the Ca2+-entry. Results obtained using pharmacological tools suggest that the contribution of Ca2+ sources to ATP-induced [Ca2+]i rise in t-BBEC117 is estimated as approximately 2:1:2 for Ca2+-release and Ca2+-entry though SOCs and ROCs, respectively. [Abstract/Link to Full Text]

Yildiz O
Vasodilating mechanisms of levosimendan: involvement of K+ channels.
J Pharmacol Sci. 2007 May;104(1):1-5.
Levosimendan, a novel agent developed for the treatment of acute and decompensated heart failure, exerts potent positive inotropic action and peripheral vasodilatory effects. The mechanism of vasodilation by levosimendan may involve reduction of Ca2+ sensitivity of contractile proteins in vascular smooth muscle, the lowering of intracellular free Ca2+, the potential inhibition of phosphodiesterase (PDE) III, and an opening of K+ channels. Although the importance and relative contribution of each of these mechanisms of vasorelaxation is unclear and may be different in various vessels and dependent on the dose of levosimendan, the important roles of K+-channel opening and Ca2+ desensitization in vascular smooth muscle are obvious, whereas the role of PDE inhibition remains to be defined. This review article briefly discusses the current research data on the mechanism of levosimendan-induced vasodilation with an emphasis on the types of the blood vessels and the K+ channels. It also summarizes the current experimental and clinical knowledge of the use of levosimedan in the treatment of heart failure. [Abstract/Link to Full Text]

Dobric I, Drvis P, Petrovic I, Shejbal D, Brcic L, Blagaic AB, Batelja L, Sever M, Kokic N, Tonkic A, Zoricic I, Mise S, Staresinic M, Radic B, Jakir A, Babel J, Ilic S, Vuksic T, Jelic I, Anic T, Seiwerth S, Sikiric P
Prolonged esophagitis after primary dysfunction of the pyloric sphincter in the rat and therapeutic potential of the gastric pentadecapeptide BPC 157.
J Pharmacol Sci. 2007 May;104(1):7-18.
Seven or fourteen days or twelve months after suturing one tube into the pyloric sphincter (removed by peristalsis by the seventh day), rats exhibit prolonged esophagitis with a constantly lowered pressure not only in the pyloric, but also in the lower esophageal sphincter and a failure of both sphincters. Throughout the esophagitis experiment, gastric pentadecapeptide BPC 157 (PL 14736) is given intraperitoneally once a day (10 microg/kg, 10 ng/kg, last application 24 h before assessment), or continuously in drinking water at 0.16 microg/ml, 0.16 ng/ml (12 ml/rat per day), or directly into the stomach 5 min before pressure assessment (a water manometer connected to the drainage port of a Foley catheter implanted into the stomach either through an esophageal or duodenal incision). This treatment alleviates i) the esophagitis (macroscopically and microscopically, at either region or interval), ii) the pressure in the pyloric sphincter, and iii) the pressure in the lower esophageal sphincter (cmH2O). In the normal rats it increases lower esophageal sphincter pressure, but decreases the pyloric sphincter pressure. Ranitidine, given using the same protocol (50 mg/kg, intraperitoneally, once daily; 0.83 mg/ml in drinking water; 50 mg/kg directly into the stomach) does not have an effect in either rats with esophagitis or in normal rats. [Abstract/Link to Full Text]

Tan-No K, Esashi A, Nakagawasai O, Niijima F, Furuta S, Sato T, Satoh S, Yasuhara H, Tadano T
Intrathecally administered D-cycloserine produces nociceptive behavior through the activation of N-methyl-D-aspartate receptor ion-channel complex acting on the glycine recognition site.
J Pharmacol Sci. 2007 May;104(1):39-45.
Intrathecal (i.t.) administration of D-cycloserine (100 and 300 fmol), a partial agonist of the glycine recognition site on the N-methyl-D-aspartate (NMDA) receptor ion-channel complex, produced a behavioral response mainly consisting of biting and/or licking of the hindpaw and the tail along with slight hindlimb scratching directed toward the flank in mice, which peaked at 5 - 10 min and almost disappeared at 15 min after the injection. The behavior induced by D-cycloserine (300 fmol) was dose-dependently inhibited by an intraperitoneal injection of morphine (0.5-2 mg/kg), suggesting that the behavioral response is related to nociception. The nociceptive behavior was also dose-dependently inhibited by i.t. co-administration of 7-chlorokynurenic acid (0.25-4 nmol), a competitive antagonist of the glycine recognition site on the NMDA receptor ion-channel complex; D-(-)-2-amino-5-phosphonovaleric acid (62.5-500 pmol), a competitive NMDA receptor antagonist; MK-801 (62.5-500 pmol), an NMDA ion-channel blocker; ifenprodil (0.5-8 nmol); arcaine (31-125 pmol); and agmatine (0.1-10 pmol), all being antagonists of the polyamine recognition site on the NMDA receptor ion-channel complex. However, [D-Phe7,D-His9]-substance P(6-11), a specific antagonist for substance P (NK1) receptors, and MEN-10,376, a tachykinin NK2-receptor antagonist, had no effect on D-cycloserine-induced nociceptive behavior. These results in the mouse spinal cord suggest that D-cycloserine-induced nociceptive behavior is mediated through the activation of the NMDA receptor ion-channel complex by acting on the glycine recognition site and that it does not involve the tachykinin receptor mechanism. [Abstract/Link to Full Text]

Ishige K, Takagi N, Imai T, Rausch WD, Kosuge Y, Kihara T, Kusama-Eguchi K, Ikeda H, Cools AR, Waddington JL, Koshikawa N, Ito Y
Role of caspase-12 in amyloid beta-peptide-induced toxicity in organotypic hippocampal slices cultured for long periods.
J Pharmacol Sci. 2007 May;104(1):46-55.
Amyloid beta (Abeta) toxicity has been implicated in cell death in the hippocampus, but its specific mechanisms are poorly understood. In this study, Abeta-induced cell death was investigated in organotypic hippocampal slice cultures (OHCs) that were cultured for various periods in vitro. There were no obvious histological differences among slices cultured for 3 to 7 weeks in vitro. Although there was little neurotoxicity after treatment with Abeta25-35 in OHCs cultured for relatively shorter periods (3-5 weeks), age-dependent cell death was evident in OHCs cultured for relatively longer periods (6-7 weeks) after exposure to Abeta25-35. In OHCs cultured for 7 weeks, S-allyl-L-cysteine (SAC), a component of aged garlic extract, protected the cells in areas CA1 and CA3 and the dentate gyrus from Abeta25-35-induced toxicity. The immunoreactivity of cleaved caspase-12 was increased whereas that of glucose-regulated protein 78 was not altered after exposure to Abeta25-35. The increases in the cleaved caspase-12 were also reversed by simultaneously applied SAC. These results suggest that OHCs cultured for relatively longer periods are more susceptible to Abeta-induced toxicity and that the Abeta-induced cell death involves caspase-12-dependent pathways. It is also suggested that SAC is able to protect against the Abeta-induced neuronal cell death through the inhibition of the caspase-12-dependent pathway. [Abstract/Link to Full Text]

Murakami A, Ishikawa T, Takechi K, Ago J, Matsumoto N, Kamei C
Effects of certain antiepileptics on behavioral and electromyographic seizure patterns induced by maximal electroshock in mice.
J Pharmacol Sci. 2007 May;104(1):56-60.
The changes of electromyographic activity (EMG seizure) induced by maximal electroshock were studied in comparison with those of behavioral seizures in mice. In addition, the effects of certain antiepileptics on behavioral seizures and EMG seizure induced by maximal electroshock were also studied. High amplitude with high frequency EMG seizure was observed in parallel with the appearance of tonic extensor (TE) seizure and an intimate relationship was observed between the two parameters. On the other hand, to investigate the intensity of TE seizure, the product of the amplitude and the duration in EMG seizure was calculated, and the effects of antiepileptics on the magnitude of EMG seizure were investigated. As a result, a significant difference was observed at the doses of antiepileptics that showed no significant effects on the durations of TE and EMG seizures; that is, phenytoin, phenobarbital, topiramate, and carbamazepine showed significant effects on the magnitude of EMG seizure at doses of 5, 2, 10, and 5 mg/kg, respectively. From these findings, it may be concluded that this index, that is, the magnitude of EMG seizure induced by maximal electroshock, is a more reliable and highly sensitive method for the assessment of the potential activity of antiepileptics. [Abstract/Link to Full Text]

Hara S, Mukai T, Mizukami H, Kuriiwa F, Watanabe T, Endo T
Nitric oxide-independent cGMP efflux in the striatum of rats exposed to carbon monoxide as determined by microdialysis.
J Pharmacol Sci. 2007 May;104(1):90-3.
Extracellular cGMP in the striatum of rats exposed to 3000 ppm carbon monoxide (CO) or 8% O2 was decreased during the early period of exposure. Thereafter, extracellular cGMP in rats exposed to CO, but not 8% O2, was transiently increased. A nitric oxide (NO) synthase inhibitor, NG-nitro-L-arginine, strongly reduced the steady-state level of extracellular cGMP in the striatum, indicating a primary role of NO in cGMP production. However, it failed to suppress the CO-induced increase in extracellular cGMP in the striatum. These findings suggest that CO may stimulate cGMP production in rat striatum independently of NO and hypoxia. [Abstract/Link to Full Text]

Morimoto T, Ohya S, Hayashi H, Onozaki K, Imaizumi Y
Cell-cycle-dependent regulation of Ca2+-activated K+ channel in Jurkat T-lymphocyte.
J Pharmacol Sci. 2007 May;104(1):94-8.
Small-conductance Ca2+-activated K+ (SK2) channel plays an important role in the activation of Jurkat T-lymphocytes by maintaining electrical gradients for the sustained Ca2+ influx. Apamin-sensitive K+ current was significantly decreased with cell-cycle progression from G0/G1 into G2/M phases, and protein expression of SK2 channels showed parallel down-regulation, with its highest expression at early G0/G1 phase. In the G0/G1 phase, the apamin-sensitive component of thapsigargin-induced Ca2+ influx was significantly larger than that in the G2/M phase. These observations suggest that SK2-channel activation may largely contribute to the sustained Ca2+ influx in the G0/G1 phase in comparison of that in the G2/M phase in Jurkat T-lymphocytes. [Abstract/Link to Full Text]

Akiyama Y, Miwa S
Improvement of postischemic dopaminergic dysfunction by edaravone, a free radical scavenger.
J Pharmacol Sci. 2007 May;104(1):99-102.
Effects of a free radical scavenger, edaravone (3-methyl-1-phenyl-2-pyrazolin-5-one), on ischemia/reperfusion-induced dysfunctions of rat striatal dopaminergic neurons were examined using in vivo brain microdialysis. During transient forebrain ischemia, dopamine levels in dialysates were elevated 140-fold above controls but rapidly recovered after reperfusion. The increase in dopamine levels induced by high K+ stimulation after reperfusion was far smaller than that of the controls. Pretreatment with edaravone but not post-treatment dose-dependently improved the response to high K+ but not the massive dopamine increase during ischemia. These results suggest that free radicals produced during ischemia play more important roles in ischemia/reperfusion-induced dysfunctions of dopaminergic neurons. [Abstract/Link to Full Text]

Dongre SH, Badami S, Natesan S, H RC
Antitumor Activity of the Methanol Extract of Hypericum hookerianum Stem Against Ehrlich Ascites Carcinoma in Swiss Albino Mice.
J Pharmacol Sci. 2007 Apr;103(4):354-9.
A large number of plants belonging to the Hypericum family are known to possess strong antitumor properties. The methanol extract of H. hookerianum Wight and Arnott stem (MEHH) exhibited potent in vitro cytotoxic activity against various cancerous cell lines. In the present study, the high performance liquid chromatography (HPLC) standardized MEHH was tested for in vivo antitumor properties against Ehrlich ascites carcinoma (EAC) tumor bearing mice at 100, 200, and 400 mg/kg body weight doses given orally once daily for 14 days. The results indicate that administration of the extract not only increased the survival of animals with ascites tumor, decreased the body weight induced by the tumor burden, and reduced packed cell volume and viable tissue cell count, but also altered many hematological parameters changed during tumor progression, indicating the potent antitumor nature of the extract. Among the three doses tested, the 200 mg/kg body weight dose was found to be the most potent. [Abstract/Link to Full Text]

Morita H, Honda A, Inoue R, Ito Y, Abe K, Nelson MT, Brayden JE
Membrane stretch-induced activation of a TRPM4-like nonselective cation channel in cerebral artery myocytes.
J Pharmacol Sci. 2007 Apr;103(4):417-26.
Stretch-activated cation channels (SACs) have been observed in many types of smooth muscle cells. However, the molecular identity and activation mechanisms of SACs remain poorly understood. We report that TRPM4-like cation channels are activated by membrane stretch in rat cerebral artery myocytes (CAMs). Negative pressure (> or =20 mmHg, cell-attached mode) activated single channels (approximately 20 pS) in isolated CAMs. These channels were permeable to Na(+) and Cs(+) and inhibited by Gd(3+) (30 microM) and DIDS (100 microM). The effect of negative pressure was abolished by membrane excision, but subsequent application of Ca(2+) (>100 nM) to the intracellular side of the membrane restored single channel activity that was indistinguishable from SACs. Caffeine (5 mM), which depletes SR Ca(2+)-stores, first activated and then abolished SACs. Tetracaine (100 microM), a ryanodine receptor antagonist, inhibited SACs. Overexpression of hTRPM4B in HEK293 cells resulted in the appearance of cation channels that were activated by both negative pressure and Ca(2+) and which had very similar biophysical and pharmacological properties as compared with SACs in CAMs. These studies indicate that TRPM4-like channels in CAMs can be activated by membrane stretch, possibly through ryanodine receptor activation, and this may contribute to the depolarization and concomitant vasoconstriction of intact cerebral arteries following mechanical stimulation. [Abstract/Link to Full Text]

Zhao Q, Murakami Y, Tohda M, Obi R, Shimada Y, Matsumoto K
Chotosan, a kampo formula, ameliorates chronic cerebral hypoperfusion-induced deficits in object recognition behaviors and central cholinergic systems in mice.
J Pharmacol Sci. 2007 Apr;103(4):360-73.
We previously demonstrated that the Kampo formula chotosan (CTS) ameliorated spatial cognitive impairment via central cholinergic systems in a chronic cerebral hypoperfusion (P2VO) mouse model. In this study, the object discrimination tasks were used to determine if the ameliorative effects of CTS on P2VO-induced cognitive deficits are a characteristic pharmacological profile of this formula, with the aim of clarifying the mechanisms by which CTS enhances central cholinergic function in P2VO mice. The cholinesterase inhibitor tacrine (THA) and Kampo formula saikokeishito (SKT) were used as controls. P2VO impaired object discrimination performance in the object recognition, location, and context tests. Daily administration of CTS (750 mg/kg, p.o.) and THA (2.5 mg/kg, i.p.) improved the object discrimination deficits, whereas SKT (750 mg/kg, p.o.) did not. In ex vivo assays, tacrine but not CTS or SKT inhibited cortical cholinesterase activity. P2VO reduced the mRNA expression of m(3) and m(5) muscarinic receptors and choline acetyltransferase but not that of other muscarinic receptor subtypes in the cerebral cortex. Daily administration of CTS and THA but not SKT reversed these expression changes. These results suggest that CTS and THA improve P2VO-induced cognitive impairment by normalizing the deficit of central cholinergic systems and that the beneficial effect on P2VO-induced cognitive deficits is a distinctive pharmacological characteristic of CTS. [Abstract/Link to Full Text]

Das AK, Yoshimura S, Mishima R, Fujimoto K, Mizuguchi H, Dev S, Wakayama Y, Kitamura Y, Horio S, Takeda N, Fukui H
Stimulation of histamine H1 receptor up-regulates histamine H1 receptor itself through activation of receptor gene transcription.
J Pharmacol Sci. 2007 Apr;103(4):374-82.
Histamine is a major mediator in allergy acting mainly through the histamine H(1) receptor (H1R). Although H1R up-regulation has been suggested as an important step for induction of allergic symptoms, little is known about the regulation of H1R level. Here we report that the activation of H1R up-regulates H1R through augmentation of H1R mRNA expression in HeLa cells. Histamine stimulation significantly increased both H1R promoter activity and mRNA level without alteration in mRNA stability. H1R protein was also up-regulated by histamine. An H1R antagonist but not histamine H(2) receptor antagonist blocked histamine-induced up-regulation of both promoter activity and mRNA expression. A protein kinase C (PKC) activator, phorbol-12-myristate-13-acetate, increased H1R mRNA expression, whereas an activator of PKA or PKG (8-Br-cAMP or 8-Br-cGMP, respectively) did not. Furthermore, histamine-induced up-regulation of both promoter activity and mRNA level were completely suppressed by the PKC inhibitor Ro-31-8220. H1R antagonists have long been thought to block H1R and inhibit immediate allergy symptoms. In addition to this short-term effect, our data propose their long-term inhibitory effect against allergic diseases by suppressing PKC-mediated H1R gene transcription. This finding provides new insights into the therapeutic target of H1R antagonist in allergic diseases. [Abstract/Link to Full Text]

Toyama T, Kudo N, Mitsumoto A, Hibino Y, Tsuda T, Kawashima Y
Stearoyl-CoA desaturase activity is elevated by the suppression of its degradation by clofibric acid in the liver of rats.
J Pharmacol Sci. 2007 Apr;103(4):383-90.
A mechanism by which fibrates control stearoyl-CoA desaturase (SCD) in the liver was studied. Treatment of rats with 2-(4-chlorophenoxy)-2-methylpropionic acid (clofibric acid) or feeding of a fat-free diet markedly elevated hepatic activity of SCD. Both the treatment with clofibric acid and the feeding of the fat-free diet caused an increase in the steady-state level of SCD1 mRNA and enhanced transcriptional rate. The half-lives of SCD for control rats, rats treated with clofibric acid rats, and rats fed the fat-free diet were estimated to be 2.0, 3.9, and 1.9 h, respectively. Activity of palmitoyl-CoA chain elongase (PCE) was increased by both clofibric acid treatment and feeding of the fat-free diet as was observed with SCD. Steady-state level of rat fatty acid elongase 2 mRNA was increased by the treatment with clofibric acid or feeding of fat-free diet, although the transcriptional rate was not altered. Different from SCD, PCE was highly stable and its half-life was not changed by either clofibric acid or fat-free diet. These results strongly suggest that the decreased degradation of SCD is responsible for the increase in its activity in addition to increased transcription of SCD1 in the rats treated with clofibric acid. [Abstract/Link to Full Text]

Nishiya D, Enomoto S, Omura T, Matsumoto R, Kusuyama T, Izumi Y, Iwao H, Takeuchi K, Yoshiyama M
The long-acting Ca2+-channel blocker azelnidipine prevents left ventricular remodeling after myocardial infarction.
J Pharmacol Sci. 2007 Apr;103(4):391-7.
Long-acting Ca(2+)-channel blockers have been reported to be effective in treating ischemic heart disease. However, their effects on cardiac remodeling after myocardial infarction (MI) are still unclear. We performed this study to examine the effect of azelnidipine on left ventricular (LV) remodeling, including systolic and diastolic dysfunction, in rats with MI. MI was induced by ligation of the left anterior descending artery. The rats were then separated into 3 groups: a sham-operated group (n = 9), untreated MI group (n = 10), and azelnidipine-treated MI group (n = 10). Four weeks after MI, hemodynamic measurements and Doppler echocardiographic assessment were performed. LV weight and LV end-diastolic dimension were significantly higher in the untreated MI group than in the sham-operated group. Azelnidipine significantly prevented the increases in these parameters. Azelnidipine also improved the ejection fraction (42 +/- 3%, P<0.05) and the E wave to A wave ratio (3.2 +/- 0.5, P<0.05), compared with the untreated MI group (31 +/- 3% and 5.3 +/- 0.8, respectively). In conclusion, azelnidipine can prevent LV remodeling and improve systolic and diastolic function after MI. Administration of long-acting Ca(2+)-channel blockers after MI is an effective strategy for treating MI. [Abstract/Link to Full Text]

Lee HM, Won KJ, Kim J, Park HJ, Kim HJ, Roh HY, Lee SH, Lee CK, Kim B
Endothelin-1 induces contraction via a Syk-mediated p38 mitogen-activated protein kinase pathway in rat aortic smooth muscle.
J Pharmacol Sci. 2007 Apr;103(4):427-33.
Although spleen tyrosine kinase (Syk) has crucial roles in various cells, its function on vascular smooth muscle contraction has not been determined. In the present study, we performed experiments to determine if Syk contributes to the endothelin-1 (ET-1)-mediated contraction in rat aortic smooth muscle. ET-1-induced contraction of aortic strips was inhibited by piceatannol, PD98059, and SB203580, inhibitors of Syk, extracellular signal-regulated kinase 1/2 (ERK1/2), and p38 mitogen-activated protein kinase (MAPK), respectively. Piceatannol also attenuated high K(+)-induced contraction. ET-1 dose-dependently enhanced the activity of Syk and this was inhibited by piceatannol in both rat aortic strip and rat aortic smooth muscle cells. The phosphorylation of p38 MAPK and heat shock protein 27 (HSP27), but not that of ERK1/2, in response to ET-1 was inhibited by both piceatannol and SB203580. These results suggest that Syk may play an important role in the regulation of aortic smooth muscle contraction induced by ET-1, which may be mediated by the p38 MAPK/HSP27 signaling pathway. [Abstract/Link to Full Text]

Hashimoto K
Arrhythmia models for drug research: classification of antiarrhythmic drugs.
J Pharmacol Sci. 2007 Apr;103(4):333-46.
The aim of this study was to classify antiarrhythmic drugs based on their effectiveness on 6 in vivo arrhythmia models, mainly using dogs. The models were produced by two-stage coronary ligation, digitalis, halothane-adrenaline, programmed electrical stimulation in old myocardial infarction dogs, coronary artery occlusion/reperfusion, or chronic atrioventricular block. Na(+)-channel-blocking drugs suppressed two-stage coronary ligation and digitalis arrhythmias. Ca(2+)-channel blockers and beta-blockers suppressed halothane-adrenaline arrhythmia. Positive inotropic drugs aggravated halothane-adrenaline arrhythmia, but did not aggravate digitalis arrhythmia. K(+)-channel blockers suppressed programmed electrical stimulation induced arrhythmia, but induced torsades de pointes type arrhythmia in chronic atrioventricular block dogs and aggravated halothane-adrenaline arrhythmia. Na(+)/H(+)-exchange blockers suppressed coronary artery occlusion/reperfusion arrhythmias. This classification may be useful for predicting the clinical effectiveness in the preclinical stage of drug development. [Abstract/Link to Full Text]

Kobayashi T, Yamada Y, Fukao M, Tsutsuura M, Tohse N
Regulation of Cav1.2 current: interaction with intracellular molecules.
J Pharmacol Sci. 2007 Apr;103(4):347-53.
Ca(V)1.2 (alpha(1c)) is a pore-forming subunit of the voltage-dependent L-type calcium channel and is expressed in many tissues. The beta and alpha(2)/delta subunits are auxiliary subunits that affect the kinetics and the expression of Ca(V)1.2. In addition to the beta and alpha(2)/delta subunits, several molecules have been reported to be involved in the regulation of Ca(V)1.2 current. Calmodulin, CaBP1 (calcium-binding protein-1), CaMKII (calcium/calmodulin-dependent protein kinase II), AKAPs (A-kinase anchoring proteins), phosphatases, Caveolin-3, beta(2)-adrenergic receptor, PDZ domain proteins, sorcin, SNARE proteins, synaptotagmin, CSN5, RGK family, and AHNAK1 have all been reported to interact with Ca(V)1.2 and the beta subunit. This review focuses on the effect of these molecules on Ca(V)1.2 current. [Abstract/Link to Full Text]

Kimura M, Okamoto H, Ogihara M
Activation of mitogen-activated protein kinase by hepatocyte growth factor is stimulated by both alpha1- and beta2-adrenergic agonists in primary cultures of adult rat hepatocytes.
J Pharmacol Sci. 2007 Apr;103(4):398-407.
We investigated the effects of alpha(1)- and beta(2)-adrenergic agonists on hepatocyte growth factor (HGF)-stimulated mitogen-activated protein kinase (MAPK) isoforms in primary cultures of adult rat hepatocytes. Hepatocytes were isolated and cultured with HGF (5 ng/ml) and/or alpha- and beta-adrenergic agonists. Phosphorylated MAPK isoforms (p42 and p44 MAPK) were detected by Western blotting analysis using anti-phospho-MAPK antibody. The results show that HGF increased phosphorylation of p42 MAPK by 2.2-fold within 3 min. The HGF-induced MAPK activation was abolished by AG1478 treatment (10(-7) M). The MEK (MAPK kinase) inhibitor PD98059 (10(-6) M) completely inhibited the HGF-dependent increase in MAPK activity. Phenylephrine (10(-6) M) and metaproterenol (10(-6) M) alone had no effect in the absence of HGF, but significantly increased p42 MAPK induction by HGF. Moreover, the cell-permeable cAMP analog, 8-bromo cAMP (10(-7) M), and phorbol 12-myristate 13 acetate (10(-7) M) potentiated HGF-induced MAPK phosphorylation. The effects of these analogs were antagonized by the protein kinase A (PKA) inhibitor H-89 (10(-7) M) and the protein kinase C (PKC) inhibitor sphingosine (10(-6) M), respectively. These results suggest that direct or indirect activation of both PKA and PKC represent a positive regulatory mechanism for stimulating MAPK induction by HGF. [Abstract/Link to Full Text]

Takayasu Y, Nakaki J, Kawasaki T, Koda K, Ago Y, Baba A, Matsuda T
Edaravone, a radical scavenger, inhibits mitochondrial permeability transition pore in rat brain.
J Pharmacol Sci. 2007 Apr;103(4):434-7.
Edaravone, a radical scavenger, prevents ischemia/reperfusion injury in the brain, but the detailed mechanism is not known. This study examines the effect of edaravone on mitochondrial permeability transition pore (PTP) in rat brain. Edaravone at 10 - 100 microM inhibited Ca(2+)- and H(2)O(2)-induced swelling of mitochondria isolated from rat brain. Addition of Ca(2+) generated reactive oxygen species (ROS) in isolated mitochondria. Edaravone (10 - 100 microM) inhibited Ca(2+)-induced generation of ROS. These results suggest that edaravone inhibits opening of mitochondrial PTP in the brain, and they imply that inhibition of mitochondrial PTP may account for the neuroprotective effect of edaravone. [Abstract/Link to Full Text]

Iwata K, Nishinaka T, Matsuno K, Kakehi T, Katsuyama M, Ibi M, Yabe-Nishimura C
The activity of aldose reductase is elevated in diabetic mouse heart.
J Pharmacol Sci. 2007 Apr;103(4):408-16.
The importance of aldose reductase (AR) has been implicated in the pathogenesis of diabetic complications, although the alterations in the expression and activity of AR during hyperglycemia in the heart have not been well characterized. We investigated the expression and enzyme activity of AR in a murine diabetic model. Three weeks after the induction of hyperglycemia with streptozotocin, the level of AR mRNA was significantly reduced in the cardiac ventricles of BDF-1 mice. In contrast, the activity of AR was significantly elevated in the heart without any significant change in the protein level. In these mice, the level of cardiac thiobarbituric acid-reactive substances was unaltered, whereas the level of reduced glutathione (GSH) was significantly increased. Daily administration of insulin for 3 weeks completely normalized the level of AR mRNA and the enzyme activity. On the other hand, daily administration of an antioxidant, N-acetylcysteine significantly reduced the level of AR mRNA in the heart with a concomitant elevation in the enzyme activity. These results suggest that the activity of AR in the heart is affected by GSH dynamics. Augmented AR activity at the early stage of hyperglycemia may perturb glycolysis and affect cardiac performance. [Abstract/Link to Full Text]

Seyrek M, Yildiz O, Ulusoy HB, Yildirim V
Testosterone relaxes isolated human radial artery by potassium channel opening action.
J Pharmacol Sci. 2007 Mar;103(3):309-16.
Preliminary clinical studies of testosterone therapy in male patients with coronary artery disease obtained promising results. However, little is known about the in vitro effects of testosterone in human isolated arteries. We investigated the effect of testosterone on contractile tone of human isolated radial artery (RA). Testosterone was added (0.1 - 300 microM ) cumulatively to organ baths after precontraction with KCl (45 mM) and phenylephrine (PE, 10 microM). Testosterone-induced relaxations were tested in the presence of the cyclooxygenase inhibitor indomethacin (10 microM), nitric oxide synthase inhibitor N(omega)-nitro-L-arginine methyl ester (L-NAME, 100 microM), non-selective large conductance Ca(2+)-activated and voltage-sensitive K(+) channel inhibitor tetraethylammonium (TEA, 1 mM), ATP-sensitive K(+) channel inhibitor glibenclamide (GLI, 10 microM), and voltage-sensitive K(+) channel inhibitor 4-aminopyridine (4-AP, 1 mM). Testosterone produced relaxation in human RA (E(max): 53.03 +/- 2.76% and 66.83 +/- 1.97% of KCl and PE-induced contraction, respectively). Except for GLI, the relaxation to testosterone is affected by neither K(+) channel inhibitors (TEA, BaCl(2), and 4-AP), L-NAME, nor indomethacin. We report for the first time that supraphysiological concentrations of testosterone induces relaxation in RA. This response may occur in part via ATP-sensitive K(+) channel opening action. [Abstract/Link to Full Text]

Xie W, Wang W, Su H, Xing D, Cai G, Du L
Hypolipidemic mechanisms of Ananas comosus L. leaves in mice: different from fibrates but similar to statins.
J Pharmacol Sci. 2007 Mar;103(3):267-74.
In this study, we investigated hypolipidemic mechanisms of the ethanolic extract of Ananas comosus L. leaves (AC) in mice and then determined its activities in related enzymes. The results showed that AC (0.40 g/kg) significantly inhibited the increase in serum triglycerides by 40% in fructose-fed mice. In mice induced by alloxan and high-fat diets, serum total cholesterol remained at a high level (180 - 220 mg/dl) within 7 days of removing high-fat diets but reached normal level (120 - 140 mg/dl) after AC (0.40 g/kg per day) treatment. Also, AC (0.40 and 0.80 g/kg) significantly inhibited serum lipids from the increase in Triton WR-1339-induced hyperlipidemic mice. AC (0.01 - 100 microg/ml) selectively activated lipoprotein lipase (LPL) activity by 200% - 400% and significantly inhibited 3-hydroxyl-methyl glutaryl coenzyme A (HMGCoA) reductase activity by 20% - 49% in vitro. Furthermore, 2 months of fenofibrate (0.20 g/kg) administration particularly increased mice liver weights (0.0760 +/- 0.0110 g/g) while AC (0.40 g/kg) had no effect (0.0403 +/- 0.0047). Taken together, these results suggest that AC will be a new potential natural product for the treatment of hyperlipidemia that exerts its actions through mechanisms of inhibiting HMGCoA reductase and activating LPL activities. Its action mechanisms differentiate from those with fibrates but may be partly similar to those with statins. It is hopeful that AC may serve as the adjuvant for fibrates. [Abstract/Link to Full Text]

Minami K, Uezono Y, Ueta Y
Pharmacological aspects of the effects of tramadol on G-protein coupled receptors.
J Pharmacol Sci. 2007 Mar;103(3):253-60.
Tramadol is an analgesic that is used worldwide, but its mechanisms of action have not been elucidated. It has been speculated that tramadol acts primarily through the activation of micro-opioid receptors and the inhibition of monoamine reuptake. The majority of studies to date have focused on ion channels in the central nervous system as targets of anesthetics and analgesics. During the past decade, major advances have been made in our understanding of the physiology and pharmacology of G-protein coupled receptor (GPCR) signaling. Several studies have shown that GPCRs and ion channels are targets for analgesics and anesthetics. In particular, tramadol has been shown to affect GPCRs, including muscarinic acetylcholine receptors and 5-hydroxytryptamine receptors. Here, the effects of tramadol on monoamine transporters, GPCRs, and ion channels are presented, and recent research on the pharmacology of tramadol is discussed. [Abstract/Link to Full Text]

Ma JL, Yang PY, Rui YC, Lu L, Kang H, Zhang J
Hemin modulates cytokine expressions in macrophage-derived foam cells via heme oxygenase-1 induction.
J Pharmacol Sci. 2007 Mar;103(3):261-6.
Lipid-laden foam cells were considered to be targets for therapeutic intervention in atherosclerosis. Several studies proposed new approaches to alter both lipid accumulation and inflammatory responses in macrophages. Finding anti-inflammatory signals during foam cell formation would provide new valid targets for anti-atherosclerotic treatment. The aim of the present study was to see whether oxidized low-density lipoprotein (ox-LDL) can active heme oxygenase (HO)-1 expression level in a human monocyte line, U937 cells, associated with the increase of cytokine secretion. We used hemin (HO-1 activator) and zinc protoporphyrin IX (ZnPP IX, HO-1 inhibitor) to determine the effect of HO-1 on the regulation of cytokine expressions. The results showed that hemin can significantly decrease pro-inflammatory cytokines interleukin (IL)-1beta and tumor necrosis factor (TNF)-alpha levels, while enhancing IL-10 production in a dose-dependent manner in U937 foam cells. ZnPP IX did not significantly affect cytokine levels in foam cells. Our present results suggested that HO-1 is an important anti-inflammatory therapeutic target through inhibiting pro-inflammatory cytokines and enhancing anti-inflammatory cytokines for the management of atherogenesis. [Abstract/Link to Full Text]

Kao TY, Huang WT, Chang CP, Lin MT
Aspirin may exert its antipyresis by inhibiting the N-methyl-D-aspartate receptor-dependent hydroxyl radical pathways in the hypothalamus.
J Pharmacol Sci. 2007 Mar;103(3):293-8.
Recent findings have suggested that the N-methyl-D-aspartate (NMDA) receptor-dependent hydroxyl radical pathway in the hypothalamus of rabbit brain may mediate the fever induced by lipopolysaccharide (LPS). The aim of this study was to investigate whether aspirin exerts its antipyresis by suppressing hypothalamic glutamate and hydroxyl radicals in rabbits. The microdialysis probes were stereotaxically and chronically implanted into the preoptic anterior hypothalamus of rabbit brain for determination of both glutamate and hydroxyl radicals in situ. It was found that intravenous (i.v.) injection of LPS, in addition to inducing fever, caused increased levels of both glutamate and hydroxyl radicals in the hypothalamus. Pretreatment with aspirin (10 - 60 mg/kg, i.v.) one hour before an i.v. dose of LPS significantly reduced the febrile response and attenuated the LPS-induced increased levels of both glutamate and hydroxyl radicals in the hypothalamus. The increased levels of prostaglandin E(2) (PGE(2)) in the hypothalamus induced by LPS could be suppressed by aspirin pretreatment. The data indicate that systemic administration of aspirin, in addition to suppressing PGE(2) production, may exert its antipyresis by inhibiting the NMDA receptor-dependent hydroxyl radical pathways in the hypothalamus during LPS fever. [Abstract/Link to Full Text]

Maeda K, Sugino H, Hirose T, Kitagawa H, Nagai T, Mizoguchi H, Takuma K, Yamada K
Clozapine prevents a decrease in neurogenesis in mice repeatedly treated with phencyclidine.
J Pharmacol Sci. 2007 Mar;103(3):299-308.
It has recently been suggested that neurogenesis in the dentate gyrus is decreased in schizophrenia and this phenomenon may contribute to the pathogenesis of the disorder. Since repeated administration of psychostimulants such as phencyclidine (PCP), MK-801, and methamphetamine (METH) induces schizophrenia-like behavioral changes in animals, we investigated whether repeated administration of these psychostimulants affects neurogenesis in the dentate gyrus of mice. Newborn cells were labeled by bromodeoxyuridine (BrdU) and detected by immunohistochemistry. Repeated administration of PCP and MK-801, but not METH, resulted in a decrease in the number of BrdU-labeled cells in the dentate gyrus. PCP-induced decrease in the number of BrdU-labeled cells was negated by co-administration of clozapine, but not haloperidol, although repeated antipsychotics treatment by themselves had no effect. Furthermore, co-administration of D-serine and glycine, but not L-serine, inhibited the PCP-induced decrease in the number of BrdU-labeled cells. These results suggest that chronic dysfunction of NMDA receptors causes a decrease in neurogenesis in the dentate gyrus. [Abstract/Link to Full Text]

Chen J, Shen H, Nagasawa Y, Mitsui K, Tsurugi K, Hashimoto K
Pravastatin inhibits arrhythmias induced by coronary artery ischemia in anesthetized rats.
J Pharmacol Sci. 2007 Mar;103(3):317-22.
We have reported that chronically administered pravastatin prevented coronary artery reperfusion-induced lethal ventricular fibrillation (VF) in anesthetized rats without lowering the serum cholesterol level. The present study was undertaken to evaluate whether pravastatin prevents ischemia-induced lethal VF, simultaneously examining myeloperoxidase (MPO) activity in ischemic myocardial tissues. Anesthetized rats were subjected to 30-min ischemia and 60-min reperfusion after chronic administration of pravastatin (0.02, 0.2, and 2 mg/kg), fluvastatin (2 and 4 mg/kg), or vehicle for 22 days, orally, once daily. ECG and blood pressure were continually recorded, and MPO was measured by a spectrophotometer. Pravastatin and fluvastatin significantly (P<0.05) decreased MPO activities, but only pravastatin decreased the incidence of ischemia-induced lethal VF. Both statins had no significant effects on body weight, blood pressure, heart rate, and QT interval as we reported earlier. Our results prove further that pravastatin has benefits to decrease cardiovascular mortality beyond its cholesterol-lowering effect. Pravastatin is more potent than fluvastatin in prevention of arrhythmias. A decrease in the neutrophil infiltration may be partly involved in the inhibitory effect of pravastatin on the ischemia-induced VF. [Abstract/Link to Full Text]

Recent Articles in Alternative Medicine Review

Monograph. Withania somnifera.
Altern Med Rev. 2004 Jun;9(2):211-4. [Abstract/Link to Full Text]

Monograph. Monascus purpureus (red yeast rice).
Altern Med Rev. 2004 Jun;9(2):208-10. [Abstract/Link to Full Text]

Herman PM, Drost LM
Evaluating the clinical relevance of food sensitivity tests: a single subject experiment.
Altern Med Rev. 2004 Jun;9(2):198-207.
A number of tests are available to identify food sensitivities. This article presents an analysis of the diagnostic value of nine different food sensitivity tests run concurrently on a healthy 33-year-old female with a previous diagnosis of environmental allergies. This case study evaluated conventional allergy tests (skin prick and serum IgE), tests of other immune-mediated reactions (serum IgG and salivary IgA), and tests that claim to measure the energetic reaction of the whole person to particular foods (kinesiology, Vega, and Carroll testing). The results of an elimination/challenge test were used as indicators of true food reactions in order to calculate the sensitivity, specificity, and positive predictive value (PPV) of each test. In a separate evaluation, the variability of results across the four tests measuring IgG was determined. Results show several tests (one of the two serum tests of IgG alone, both serum tests of IgE and IgG, skin prick testing, and Carroll testing) may have very high (100 percent) specificity and PPV when test results are compared to the results of an elimination/challenge test. Sensitivity, however, is low across tests (50-60 percent), likely because different tests measure different mechanisms of food reactions and because food sensitivities can be the result of a number of different mechanisms. Very little consistency was found among the results of the four tests measuring IgG - 79-83 percent disagreement. This study shows a number of tests may be useful in identifying foods to which a patient is reactive; however, no one test is likely to identify all reactive foods. [Abstract/Link to Full Text]

Gaby AR
Single-subject experiment evaluating nine different food sensitivity tests.
Altern Med Rev. 2004 Sep;9(3):237-8; author reply 238. [Abstract/Link to Full Text]

Hawrelak JA, Myers SP
The causes of intestinal dysbiosis: a review.
Altern Med Rev. 2004 Jun;9(2):180-97.
Alterations in the bowel flora and its activities are now believed to be contributing factors to many chronic and degenerative diseases. Irritable bowel syndrome, inflammatory bowel disease, rheumatoid arthritis, and ankylosing spondylitis have all been linked to alterations in the intestinal microflora. The intestinal dysbiosis hypothesis suggests a number of factors associated with modern Western living have a detrimental impact on the microflora of the gastrointestinal tract. Factors such as antibiotics, psychological and physical stress, and certain dietary components have been found to contribute to intestinal dysbiosis. If these causes can be eliminated or at least attenuated then treatments aimed at manipulating the microflora may be more successful [Abstract/Link to Full Text]

Gaby AR
Sub-laboratory hypothyroidism and the empirical use of Armour thyroid.
Altern Med Rev. 2004 Jun;9(2):157-79.
Evidence is presented that many people have hypothyroidism undetected by conventional laboratory thyroid-function tests, and cases are reported to support the empirical use of Armour thyroid. Clinical evaluation can identify individuals with sub-laboratory hypothyroidism who are likely to benefit from thyroid-replacement therapy. In a significant proportion of cases, treatment with thyroid hormone has resulted in marked improvement in chronic symptoms that had failed to respond to a wide array of conventional and alternative treatments. In some cases, treatment with desiccated thyroid has produced better clinical results than levothyroxine. Research supporting the existence of sub-laboratory hypothyroidism is reviewed, and the author's clinical approach to the diagnosis and treatment of this condition is described. [Abstract/Link to Full Text]

Marshall K
Therapeutic applications of whey protein.
Altern Med Rev. 2004 Jun;9(2):136-56.
Whey, a protein complex derived from milk, is being touted as a functional food with a number of health benefits. The biological components of whey, including lactoferrin, beta-lactoglobulin, alpha-lactalbumin, glycomacropeptide, and immunoglobulins, demonstrate a range of immune-enhancing properties. In addition, whey has the ability to act as an antioxidant, antihypertensive, antitumor, hypolipidemic, antiviral, antibacterial, and chelating agent. The primary mechanism by which whey is thought to exert its effects is by intracellular conversion of the amino acid cysteine to glutathione, a potent intracellular antioxidant. A number of clinical trials have successfully been performed using whey in the treatment of cancer, HIV, hepatitis B, cardiovascular disease, osteoporosis, and as an antimicrobial agent. Whey protein has also exhibited benefit in the arena of exercise performance and enhancement. [Abstract/Link to Full Text]

Kidd PM
Bipolar disorder and cell membrane dysfunction. Progress toward integrative management.
Altern Med Rev. 2004 Jun;9(2):107-35.
Bipolar disorder (BD) is characterized by periods of abnormally elevated mood (mania) that cycle with abnormally lowered mood (depression). Multiple structural, metabolic, and biochemical abnormalities are evident in the brain's cortex, subcortex, and deeper regions. This disorder is highly genetically conditioned but also highly susceptible to environmental stressors: prenatal or perinatal insults, childhood sexual or physical abuse, challenging life events, substance abuse, and other toxic chemical exposures. Its high morbidity, lost productivity, and suicide risk place a great toll on society. Since World War II, BD has been steadily worsening with earlier age of onset, greater intensity of symptoms, and development of drug resistance. Incidence in children is rising and misdiagnosis is common. Disciplined management of the many risk factors is essential, including cognitive psychotherapy and support from family and community. Lithium has been the foundational treatment, followed by valproate and other mood stabilizers, antidepressants, and anticonvulsants. Several single-nutrient and multinutrient supplements have also proven beneficial. Controlled, double-blind trials show multinutrient combinations of vitamins, minerals, orthomolecules, herbals, and the omega-3 fatty acids EPA and DHA to be effective monotherapy. The molecular action of lithium and valproate converge with nutrients on the level of the cell membrane and its molecular signal transduction systems. This emergent, unified rationale presages effective integrative management of bipolar disorder. [Abstract/Link to Full Text]

Bacopa monniera. Monograph.
Altern Med Rev. 2004 Mar;9(1):79-85. [Abstract/Link to Full Text]

Gamma-linolenic acid (GLA). Monograph.
Altern Med Rev. 2004 Mar;9(1):70-8. [Abstract/Link to Full Text]

Udani J, Hardy M, Madsen DC
Blocking carbohydrate absorption and weight loss: a clinical trial using Phase 2 brand proprietary fractionated white bean extract.
Altern Med Rev. 2004 Mar;9(1):63-9.
Background: Phase 2' starch neutralizer brand bean extract product ("Phase 2") is a water-extract of a common white bean (Phaseolus vulgaris) that has been shown in vitro to inhibit the digestive enzyme alpha-amylase. Inhibiting this enzyme may prevent the digestion of complex carbohydrates, thus decreasing the number of carbohydrate calories absorbed and potentially promoting weight loss. Methods: Fifty obese adults were screened to participate in a randomized, double-blind, placebo-controlled study evaluating the effects of treatment with Phase 2 versus placebo on weight loss. Participants were randomized to receive either 1500 mg Phase 2 or an identical placebo twice daily with meals. The active study period was eight weeks. Thirty-nine subjects completed the initial screening process and 27 subjects completed the study. Results: The results after eight weeks demonstrated the Phase 2 group lost an average of 3.79 lbs (average of 0.47 lb per week) compared with the placebo group, which lost an average of 1.65 lbs (average of 0.21 lb per week), representing a difference of 129 percent (p=0.35). Triglyceride levels in the Phase 2 group were reduced an average of 26.3 mg/dL, more than three times greater a reduction than observed in the placebo group (8.2 mg/dL) (p=0.07). No adverse events during the study were attributed to the study medication. Conclusion: Clinical trends were identified for weight loss and a decrease in triglycerides, although statistical significance was not reached. Phase 2 shows potential promise as an adjunct therapy in the treatment of obesity and hypertriglyceridemia and further studies with larger numbers of subjects are warranted to conclusively demonstrate effectiveness. [Abstract/Link to Full Text]

Danesch UC
Petasites hybridus (Butterbur root) extract in the treatment of asthma--an open trial.
Altern Med Rev. 2004 Mar;9(1):54-62.
The efficacy and tolerability of a butterbur root extract (Petadolex) for the treatment of asthma was analyzed in a prospective, non-randomized, open trial. Subjects included 64 adults and 16 children/adolescents treated for two months with the extract, followed by two months during which the intake of the extract was optional. Concomitant asthma medication was permitted. The number, duration, and severity of asthma attacks decreased, while peak flow, forced expiratory volume (FEV1), and all measured symptoms improved during therapy. In addition, more than 40 percent of patients using asthma medications at baseline reduced intake of these medications by the end of the study. This study suggests the Petasites hybridus extract Petadolex is an effective and safe therapy for the treatment of asthma. [Abstract/Link to Full Text]

Osiecki H
The role of chronic inflammation in cardiovascular disease and its regulation by nutrients.
Altern Med Rev. 2004 Mar;9(1):32-53.
Multiple risk markers for atherosclerosis and cardiovascular disease act in a synergistic way through inflammatory pathways. This article discusses some of the key inflammatory biochemical risk markers for cardiovascular disease; in particular, the role of three basic cell types affected by these risk markers (endothelial cells, smooth muscle cells, and immune cells), the crucial role of inflammatory mediators, nitric oxide balance in cardiovascular pathology, and the use of nutrients to circumvent several of these inflammatory pathways. Most risk markers for cardiovascular disease have a pro-inflammatory component, which stimulates the release of a number of active molecules such as inflammatory mediators, reactive oxygen species, nitric oxide, and peroxynitrite from endothelial, vascular smooth muscle, and immune cells in response to injury. Nitric oxide plays a pivotal role in preventing the progression of atherosclerosis through its ability to induce vasodilation, suppress vascular smooth muscle proliferation, and reduce vascular lesion formation. Nutrients such as arginine, antioxidants (vitamins C and E, lipoic acid, glutathione), and enzyme cofactors (vitamins B2 and B3, folate, and tetrahydrobiopterin) help to elevate nitric oxide levels and may play an important role in the management of cardiovascular disease. Other dietary components such as DHA/EPA from fish oil, tocotrienols, vitamins B6 and B12, and quercetin contribute further to mitigating the inflammatory process. [Abstract/Link to Full Text]

Conant R, Schauss AG
Therapeutic applications of citicoline for stroke and cognitive dysfunction in the elderly: a review of the literature.
Altern Med Rev. 2004 Mar;9(1):17-31.
Citicoline (CDP-choline; cytidine 5'-diphosphocholine), a form of the essential nutrient choline, shows promise of clinical efficacy in elderly patients with cognitive deficits, inefficient memory, and early-stage Alzheimer's disease. Citicoline has also been investigated as a therapy in stroke patients, although the results of trials to date are inconclusive. Produced endogenously, citicoline serves as a choline donor in the metabolic pathways for biosynthesis of acetylcholine and neuronal membrane phospholipids, chiefly phosphatidylcholine. The principal components of citicoline, choline and cytidine, are readily absorbed in the GI tract and easily cross the blood-brain barrier. Exogenous citicoline, as the sodium salt, has been researched in animal experiments and human clinical trials that provide evidence of its cholinergic and neuroprotective actions. As a dietary supplement, citicoline appears useful for improving both the structural integrity and functionality of the neuronal membrane that may assist in membrane repair. This review, while not intended to be exhaustive, highlights the published, peer-reviewed research on citicoline with brief discussions on toxicology and safety, mechanisms of action, and pharmacokinetics. [Abstract/Link to Full Text]

McKay D
Nutrients and botanicals for erectile dysfunction: examining the evidence.
Altern Med Rev. 2004 Mar;9(1):4-16.
Erectile dysfunction affects 50 percent of men ages 40-70 in the United States and is considered an important public health problem by the National Institutes of Health. Consumers are exposed to a plethora of natural products claiming to restore erection and sexual vitality. A review of the available empirical evidence reveals most naturally occurring compounds lack adequate clinical trials to support efficacy. However, arginine, yohimbine, Panax ginseng, Maca, and Ginkgo biloba all have some degree of evidence they may be helpful for erectile dysfunction. Improvements in penile endothelial L-arginine-nitric oxide activity appear to be a unifying explanation for the actions of these naturally occurring agents. [Abstract/Link to Full Text]

Czap A
A patented double dilemma.
Altern Med Rev. 2004 Mar;9(1):3. [Abstract/Link to Full Text]

Oligomeric proanthocyanidins (OPCs). Monograph.
Altern Med Rev. 2003 Nov;8(4):442-50. [Abstract/Link to Full Text]

Methylsulfonylmethane (MSM). Monograph.
Altern Med Rev. 2003 Nov;8(4):438-41. [Abstract/Link to Full Text]

Marshall K
Interstitial cystitis: understanding the syndrome.
Altern Med Rev. 2003 Nov;8(4):426-37.
Interstitial cystitis (IC) is a chronic pain syndrome that affects close to a million people in the United States. The syndrome presents differently in many individuals, with the unifying factor being chronic pelvic pain and disruption of daily life activities. Many etiologies have been proposed as causative factors for IC, although it is likely triggered by more than one process. Treatment for many individuals revolves around symptom management and improving quality of life; however, it is imperative to remove aggravating factors such as food and daily stressors. Treatment will vary for individuals, as symptoms and etiology will differ. This article discusses nutritional and other non-toxic approaches to treating IC. [Abstract/Link to Full Text]

Logan AC
Neurobehavioral aspects of omega-3 fatty acids: possible mechanisms and therapeutic value in major depression.
Altern Med Rev. 2003 Nov;8(4):410-25.
Omega-3 fatty acids have been the subject of volumes of international research, the results of which indicate these substances may have therapeutic value in a number of medical conditions. An emerging area of research is examining the neurobehavioral aspects of omega-3 fatty acids (alpha-linolenic, eicosapentaenoic, docosahexaenoic) and the critical role of these essential fats in the functioning of the central nervous system. Investigations have linked omega-3 fatty acids to a number of neuropsychiatric disorders, including depression. The purpose of this article is to examine the possible mechanisms of action and potential clinical value of omega-3 fatty acids in major depression. A novel mechanism involving omega-3 modulation of cAMP response element binding protein (CREB) and brain-derived neurotrophic factor (BDNF) is proposed. [Abstract/Link to Full Text]

Williams JE
Portal to the interior: viral pathogenesis and natural compounds that restore mucosal immunity and modulate inflammation.
Altern Med Rev. 2003 Nov;8(4):395-409.
Most antigens, particularly viruses, enter the body through the mucosal epithelia where they are carried by afferent lymphatics to regional lymph nodes for presentation to the immune system. Although they share immunological similarities, immune processes that protect the mucosa are distinct from innate and acquired immunity. The barrier formed by the intestinal mucosa is the most studied, with its microenvironment having a marked influence on both local and systemic immune responses. A healthy microenvironment and resilient neighboring tissue provide protection against inflammation known to dampen mucosal immunity, promote carcinogenesis, contribute to systemic inflammatory processes, and result in autoimmune diseases. Numerous natural substances improve this microenvironment and thereby enhance immunity against microbial infections. Since mucosal immunity forms the first line of defense against many commonly transmitted pathogens, restoring and maintaining mucosal immunity is critical for disease prevention and intervention. This article discusses the nature of mucosal immunity and its relationship to viral infections and other conditions, and reviews natural compounds that help restore mucosal immunity. [Abstract/Link to Full Text]

Kelly GS
Bovine colostrums: a review of clinical uses.
Altern Med Rev. 2003 Nov;8(4):378-94.
Bovine colostrums are the "early" milk produced by cows during the first several days post-parturition. This "early" milk has a nutrient profile and immunological composition that differs substantially from "mature" milk. Included in the nutrient profile are higher amounts of immunoglobulins, growth factors, cytokines, and nucleosides than are found in milk. Bovine colostrums are also rich in oligosaccharides, antimicrobials, and immune-regulating factors. Available evidence suggests a beneficial effect of supplementation of bovine colostrums in improving body composition, aspects of athletic performance, diarrhea in persons with immune-deficiency syndromes, NSAID-induced gastrointestinal disturbances, and aspects of the acute phase response that occurs secondary to surgery. Specific hyperimmune bovine colostrums, produced to have high neutralizing titer activity against Cryptosporidia, H. pylori, measles, rotavirus, and Shigella sp., appear to have clinical utility in conditions associated with these infectious organisms. [Abstract/Link to Full Text]

MacKay D, Miller AL
Nutritional support for wound healing.
Altern Med Rev. 2003 Nov;8(4):359-77.
Healing of wounds, whether from accidental injury or surgical intervention, involves the activity of an intricate network of blood cells, tissue types, cytokines, and growth factors. This results in increased cellular activity, which causes an intensified metabolic demand for nutrients. Nutritional deficiencies can impede wound healing, and several nutritional factors required for wound repair may improve healing time and wound outcome. Vitamin A is required for epithelial and bone formation, cellular differentiation, and immune function. Vitamin C is necessary for collagen formation, proper immune function, and as a tissue antioxidant. Vitamin E is the major lipid-soluble antioxidant in the skin; however, the effect of vitamin E on surgical wounds is inconclusive. Bromelain reduces edema, bruising, pain, and healing time following trauma and surgical procedures. Glucosamine appears to be the rate-limiting substrate for hyaluronic acid production in the wound. Adequate dietary protein is absolutely essential for proper wound healing, and tissue levels of the amino acids arginine and glutamine may influence wound repair and immune function. The botanical medicines Centella asiatica and Aloe vera have been used for decades, both topically and internally, to enhance wound repair, and scientific studies are now beginning to validate efficacy and explore mechanisms of action for these botanicals. To promote wound healing in the shortest time possible, with minimal pain, discomfort, and scarring to the patient, it is important to explore nutritional and botanical influences on wound outcome. [Abstract/Link to Full Text]

Czap A
"Fish on Steroids"?
Altern Med Rev. 2003 Nov;8(4):358. [Abstract/Link to Full Text]

Lycopene. Monograph.
Altern Med Rev. 2003 Aug;8(3):336-42. [Abstract/Link to Full Text]

Zingiber officinale (ginger). Monograph.
Altern Med Rev. 2003 Aug;8(3):331-5. [Abstract/Link to Full Text]

Harding KL, Judah RD, Gant C
Outcome-based comparison of Ritalin versus food-supplement treated children with AD/HD.
Altern Med Rev. 2003 Aug;8(3):319-30.
Twenty children with attention deficit/hyperactivity disorder (AD/HD) were treated with either Ritalin (10 children) or dietary supplements (10 children), and outcomes were compared using the Intermediate Visual and Auditory/Continuous Performance Test (IVA/CPT) and the WINKS two-way analysis of variance with repeated measures and with Tukey multiple comparisons. Subjects in both groups showed significant gains (p less than 0.01) on the IVA/CPT's Full Scale Response Control Quotient and Full Scale Attention Control Quotient (p less than 0.001). Improvements in the four sub-quotients of the IVA/CPT were also found to be significant and essentially identical in both groups: Auditory Response Control Quotient (p less than 0.001), Visual Response Control Quotient (p less than 0.05), Auditory Attention Quotient (p less than 0.001), and Visual Attention Quotient (p less than 0.001). Numerous studies suggest that biochemical heterogeneous etiologies for AD/HD cluster around at least eight risk factors: food and additive allergies, heavy metal toxicity and other environmental toxins, low-protein/high-carbohydrate diets, mineral imbalances, essential fatty acid and phospholipid deficiencies, amino acid deficiencies, thyroid disorders, and B-vitamin deficiencies. The dietary supplements used were a mix of vitamins, minerals, phytonutrients, amino acids, essential fatty acids, phospholipids, and probiotics that attempted to address the AD/HD biochemical risk factors. These findings support the effectiveness of food supplement treatment in improving attention and self-control in children with AD/HD and suggest food supplement treatment of AD/HD may be of equal efficacy to Ritalin treatment. [Abstract/Link to Full Text]

Lamson DW, Plaza SM
The anticancer effects of vitamin K.
Altern Med Rev. 2003 Aug;8(3):303-18.
Vitamin K, an essential nutrient often associated with the clotting cascade, has been the focus of considerable research demonstrating an anticancer potential. Much of this research has focused on vitamin K3, although vitamins K2 and K1 have also been shown to have anticancer effects. Early studies of vitamin K3 employed an oxidative model to explain the anticancer effects seen in both in vitro and in vivo studies; however, this model does not adequately address the action of vitamins K1 and K2. Recent research has demonstrated the anticancer action of vitamin K may act at the level of tyrosine kinases and phosphatases, modulating various transcription factors such as Myc and Fos. Tyrosine kinases associated with cyclins have also been shown to be affected by vitamin K, which can lead to cell cycle arrest and cell death. [Abstract/Link to Full Text]

Philp HA
Hot flashes--a review of the literature on alternative and complementary treatment approaches.
Altern Med Rev. 2003 Aug;8(3):284-302.
Hot flashes are a common experience for menopausal women, with an 85-percent incidence in the West. With the increased knowledge of side effects attributable to conventional treatment options, more women are exploring natural alternatives. Although more definitive research is necessary, several natural therapies show promise in treating hot flashes without the risks associated with conventional therapies. Soy and other phytoestrogens, black cohosh, evening primrose oil, vitamin E, the bioflavonoid hesperidin with vitamin C, ferulic acid, acupuncture treatment, and regular aerobic exercise have been shown effective in treating hot flashes in menopausal women. [Abstract/Link to Full Text]

Head KA, Jurenka JS
Inflammatory bowel disease Part 1: ulcerative colitis--pathophysiology and conventional and alternative treatment options.
Altern Med Rev. 2003 Aug;8(3):247-83.
Ulcerative colitis (UC), a subcategory of inflammatory bowel disease, afflicts 1-2 million people in the United States, and many more worldwide. Although the exact cause of ulcerative colitis remains undetermined, the condition appears to be related to a combination of genetic and environmental factors. While conventional treatments can be effective in maintaining remission and decreasing the length of active disease periods, the treatments are not without side effects, and a significant number of people suffering from UC fail to respond to even the strongest drugs. This article reviews potential unconventional treatments - transdermal nicotine, heparin, melatonin, DHEA, probiotics, fiber, dietary changes, botanicals, essential fatty acids, and other nutrients - that may be considered in conjunction with conventional approaches or as part of a comprehensive alternative treatment protocol. In addition this review addresses risk factors, pathogenesis, nutrient deficiencies, conventional treatment approaches, and extra-intestinal manifestations of the disease. [Abstract/Link to Full Text]

Kidd P
Th1/Th2 balance: the hypothesis, its limitations, and implications for health and disease.
Altern Med Rev. 2003 Aug;8(3):223-46.
One theory of immune regulation involves homeostasis between T-helper 1 (Th1) and T-helper 2 (Th2) activity. The Th1/Th2 hypothesis arose from 1986 research suggesting mouse T-helper cells expressed differing cytokine patterns. This hypothesis was adapted to human immunity, with Th1- and Th2-helper cells directing different immune response pathways. Th1 cells drive the type-1 pathway ("cellular immunity") to fight viruses and other intracellular pathogens, eliminate cancerous cells, and stimulate delayed-type hypersensitivity (DTH) skin reactions. Th2 cells drive the type-2 pathway ("humoral immunity") and up-regulate antibody production to fight extracellular organisms; type 2 dominance is credited with tolerance of xenografts and of the fetus during pregnancy. Overactivation of either pattern can cause disease, and either pathway can down-regulate the other. But the hypothesis has major inconsistencies; human cytokine activities rarely fall into exclusive pro-Th1 or -Th2 patterns. The non-helper regulatory T cells, or the antigen-presenting cells (APC), likely influence immunity in a manner comparable to Th1 and Th2 cells. Many diseases previously classified as Th1 or Th2 dominant fail to meet the set criteria. Experimentally, Th1 polarization is readily transformed to Th2 dominance through depletion of intracellular glutathione, and vice versa. Mercury depletes glutathione and polarizes toward Th2 dominance. Several nutrients and hormones measurably influence Th1/Th2 balance, including plant sterols/sterolins, melatonin, probiotics, progesterone, and the minerals selenium and zinc. The long-chain omega-3 fatty acids EPA (eicosapentaenoic acid) and DHA (docosahexaenoic acid) significantly benefit diverse inflammatory and autoimmune conditions without any specific Th1/Th2 effect. Th1/Th2-based immunotherapies, e.g., T-cell receptor (TCR) peptides and interleukin-4 (IL-4) injections, have produced mixed results to date. [Abstract/Link to Full Text]

Czap A
Pass the tablet please.
Altern Med Rev. 2003 Aug;8(3):222. [Abstract/Link to Full Text]

Recent Articles in Journal of Physiology and Pharmacology

Dobrzy? A, Dobrzy? P
Stearoyl-CoA desaturase--a new player in skeletal muscle metabolism regulation.
J Physiol Pharmacol. 2006 Nov;57 Suppl 1031-42.
Stearoyl-CoA desaturase (SCD) is a rate-limiting enzyme catalyzing the synthesis of monounsaturated fatty acids, mainly oleate (18:1) and palmitoleate (16:1), which are a major component of tissue lipids. SCD1 deficient mice reveal increased energy expenditure and decreased body adiposity due to the upregulation of genes of fatty acid oxidation and the downregulation of genes of lipid synthesis in liver. In this review, we examine data showing that SCD is an important component in the regulation of skeletal muscle metabolism, which affects insulin sensitivity, mitochondrial fatty acid oxidation and ceramide de novo synthesis in oxidative myofibers. The lack of SCD1 gene increases the rate of fatty acid beta-oxidation through activation of the AMP-activated protein kinase (AMPK) pathway and by upregulating genes of fatty acid oxidation in soleus and red gastrocnemius muscles. Consistent with increased beta-oxidation, the contents of free fatty acids and long-chain acyl-CoAs are significantly decreased, which together with reduced mRNA level and activity of serine palmitoyltransferase led to reduced ceramide synthesis in oxidative muscles of SCD1-/- mice. Thus, reduced contents of free fatty acids, acyl-CoAs and ceramides as well as increased AMPK phosphorylation, might contribute to increased insulin sensitivity observed in muscle of SCD1-/- mice. SCD1 deficiency also results in downregulation of the expression of the protein-tyrosine phosphatase 1B, which is responsible for the sustained insulin receptor autophosphorylation despite reduced levels of plasma insulin in the SCD1-/- mice. SCD1 deficiency reduced ceramide synthesis, increased AMPK phosphorylation and carnitine palmitoyltransferase 1 activity also in soleus and red gastrocnemius muscles of leptin deficient ob/ob mice. These findings raise the possibility that SCD1 may be a downstream component of the leptin signaling pathway in skeletal muscle. [Abstract/Link to Full Text]

Hancock CR, Brault JJ, Terjung RL
Protecting the cellular energy state during contractions: role of AMP deaminase.
J Physiol Pharmacol. 2006 Nov;57 Suppl 1017-29.
AMP deaminase activity (AMP->IMP+NH3) is the entry reaction to the purine nucleotide cycle. In skeletal muscle, excessive energy demands during contractions leads to a net production of ADP, because ATP hydrolysis exceeds ADP rephosphorylation. Elevations in ADP increase AMP, via the myokinase reaction. This accumulation of ATP hydrolysis products should lead to a catastrophic reduction in the energy state of the myocyte. The removal of AMP to IMP in times of excessively high energy demands have been hypothesized as essential to protect the energy state of the cell. While AMP deamination leads to a net loss of adenine nucleotides (principally, as ATP), the viability of the myocyte is preserved. Following these demanding contraction conditions, the concentration of IMP of fast-twitch muscle is rapidly reduced, typically with the return of the muscle adenine nucleotide content (ATP + ADP + AMP) to pre-contraction levels. While these observations are generally observed for fast-twitch skeletal muscle and consistent with the hypothesis, there has been no direct experimental evaluation. In the AK1 (-/-) mouse, there is a markedly reduced accumulation of AMP, during conditions of excessive contractile activity. Rather, there is a high ADP concentration, approaching 1.5 mM, that remains unbound 'free' within the muscle. This contributes to an inordinate reduction in the ATP/ADP ratio. At the same time, PCr hydrolysis is nearly complete leading to a large increase in orthophosphate. In combination, this leads to an exceptional decline in the free energy of ATP hydrolysis. This is projected to impair Ca(2+) handling by the sarcoplasmic reticulum and slow cross-bridge cycling rate. The outcome should be slowed contraction characteristics and possible contracture. While some contractile changes were observed, there was a remarkable ability of the muscle to function under these challenging energetic conditions. Thus, it is not essential that the AMP deaminase reaction be operating during intense contraction conditions. This helps explain why patients deficient in AMP deaminase do not always exhibit an impaired muscle function. [Abstract/Link to Full Text]

Sargeant AJ, de Haan A
Human muscle fatigue: the significance of muscle fibre type variability studied using a micro-dissection approach.
J Physiol Pharmacol. 2006 Nov;57 Suppl 105-16.
During human locomotion the ability to generate and sustain mechanical power output is dependent on the organised variability in contractile and metabolic properties of the muscle fibres that comprise the active muscles. In studies of human exercise we have used a micro-dissection technique to obtain fragments of single muscle fibres from needle biopsies before and after exercise. Each fibre fragment is divided into two parts. One part is used to characterize the fibre type in respect of the heavy chain myosin isoform expressed. The other part of the fragment is analysed for high energy phosphate concentrations. Fibres are classified on the basis of expressing either type I, type IIA, or type IIX myosin heavy chain isoforms. It should be noted however that in the type II population many fibres co-express both IIA and the IIX isoforms and we therefore characterize these fibres on the basis of the degree of co-expression. We have used this technique to examine the time course of high energy phosphate concentration and fatigue in different fibre populations during exercise. The progressive reduction of power during maximal sprint efforts may be interpreted as the cumulative effect of metabolic depletion in successive fibre type populations from IIX to IIXa to IIAx to IIA to I. One important application of the micro-dissection technique is that PCr content may also be used as a very sensitive metabolic marker for fibre type recruitment during very short duration concentric, isometric and eccentric exercise. [Abstract/Link to Full Text]

Zwoli?ska-Wcis?o M, Brzozowski T, Mach T, Budak A, Trojanowska D, Konturek PC, Pajdo R, Drozdowicz D, Kwiecie? S
Are probiotics effective in the treatment of fungal colonization of the gastrointestinal tract? Experimental and clinical studies.
J Physiol Pharmacol. 2006 Nov;57 Suppl 935-49.
The influence of fungal colonization and probiotic treatment on the course of gastric ulcer (GU) and ulcerative colitis (UC) was not explored. Our studies included: 1) clinical investigation of 293 patients with dyspeptic and ulcer complaints and 72 patients with lower gastrointestinal (GI) tract: 60 patients with UC, 12 with irritable bowel syndrome (IBS) - the control group. Significant fungal colonization (SFC), over 10(5) CFU/ml was evaluated. Mycological investigation was performed, including qualitative and quantitative examination, according to Muller method, 2) experimental studies in rats included estimation of the influence of inoculation of Candida isolated from human GI tract on the healing process of GU, induced by acetic acid with or without probiotic Lactobacillus acidophilus (10(6) CFU/ml) introduced intragastrically (i.g.). At 0, 4, 15 and 25 day after ulcer induction. Weight, damage area, gastric blood flow (GBF) (H2 clearance), expression of mRNA for cytokines IL-beta, TNF-alpha (ELISA) were evaluated. Mycology: qualitative and quantitative examination was performed. MPO serum activity was measured. Results of clinical studies: 1) SFC was more frequent in patients with GU: 54.2% of cases and patients with over 5 years history of UC: 33.3% cases. 2) SFC delayed GU healing and influenced the maintenance of clinical symptoms in both diseases. Results of animal studies: 3) In Candida inoculated rats, the GBF was significantly lower than in the vehicle controls (saline administered group). Upregulation of TNF-alpha, IL-1 beta was recorded. The GUs were still present till 25 day in all rats inoculated with Candida, in contrast to vehicle group (reduction of ulcer in 92% at day 25). Conclusions: 1) Fungal colonization delays process of ulcer and inflammation healing of GI tract mucosa. That effect was attenuated by probiotic therapy. 2) Probiotic therapy seems to be effective in treatment of fungal colonization of GI tract. 3) Lactobacillus acidophilus therapy shortens the duration of fungal colonization of mucosa (enhanced Candida clearance is associated with IL-4, INF-gamma response). [Abstract/Link to Full Text]

Mach T
Clinical usefulness of probiotics in inflammatory bowel diseases.
J Physiol Pharmacol. 2006 Nov;57 Suppl 923-33.
Probiotics are live nonpathogenic bacteria or bacterial components that may be helpful in the prevention and treatment of acute diarrhoea in adults and children and have some effects on the course of inflammatory bowel diseases (IBD). Many experimental and clinical studies suggest that intestinal bacterial flora plays an important role in the pathogenesis of IBD, and manipulation of the luminal contents with antibiotics or probiotics represents a potentially effective therapeutic option. The beneficial effect of probiotics was demonstrated mainly in the prevention and treatment of pouchitis and in maintaining remission of mild to moderate ulcerative colitis. Probiotics seems to be less effective in patients with Crohn's disease. Randomized clinical trials are still required to further define the role of probiotics as preventive and therapeutic agents. This review summarizes the current data about probiotics in IBD. [Abstract/Link to Full Text]

Gackowska L, Michalkiewicz J, Krotkiewski M, Helmin-Basa A, Kubiszewska I, Dzierzanowska D
Combined effect of different lactic acid bacteria strains on the mode of cytokines pattern expression in human peripheral blood mononuclear cells.
J Physiol Pharmacol. 2006 Nov;57 Suppl 913-21.
The balance between immunogenic and tolerogenic activities in human immune system strongly depends on microflora-induced pro-and anti-inflammatory activities. Lactic acid bacteria (LAB) are important components of microflora. The interactions of the different strains of LAB and the cells of immune system are largely unknown. To assess if LAB strains composition would have an effect on the cellular responses profile (proliferation, cytokines synthesis) peripheral blood mononuclear cells (PBMC) model system was used. PBMC were induced by three different strains of LAB: Lactobacillus acidophilus, Lactobacillus delbrueckii spp. bulgaricus, Bifidobacterium bifidum. Tested strains were mixed together, in combinations with each other (pairs) or alone. Both, the LAB mixture as well as the pairs and the single LAB strains induced low lymphocyte proliferation (about 10% of ConA-induced response). However, the single LAB strains and their combinations were quite different cytokines inducers. First, L. acidophilus was much stronger IFN-gamma inducer than the LAB mixture, being a few times higher IL-12 stimulator than L. bulgaricus and B. bifidum. Second, L. bulgaricus and B. bifidum suppressed L.acidophilus-induced IFN-gamma synthesis to the level equal to that induced by the LAB mixture, limiting IL-12 production by about 30% and 70%, respectively. Third, the LAB strains were good IL-10 and TNF-alpha inducers, irrespectively of their combinations used. We conclude that LAB strains' pro or anti-inflammatory potentials are at least in part dependent on their composition. Low LAB mixture-induced IL-12 and IFN-gamma production and relatively high IL-10 and TNF-alpha expression may represent cellular activities normally induced in vivo by a combined action of bacterial antigens. Their presence is important to limit pro-inflammatory reactions (via IL-10) and to provide protection against infections (via TNF-alpha). [Abstract/Link to Full Text]

Heczko PB, Strus M, Kochan P
Critical evaluation of probiotic activity of lactic acid bacteria and their effects.
J Physiol Pharmacol. 2006 Nov;57 Suppl 95-12.
Probiotics discussed in this paper are evaluated using the WHO/FAO definition from 2001. The authors present a brief description of the normal microbiota of the gastrointestinal tract, discuss probiotics in the aspects of gut immunity and then move to selection of bacterial strains as probiotics. The main issue raised is the critical evaluation of probiotics in randomized clinical trials for conditions such as: infectious diarrhoea; antibiotic associated diarrhoea; inflammatory bowel disease; pouchitis and diverticulitis; H. pylori infection; irritable bowel syndrome. Safety of probiotics is mentioned with respect to susceptible individuals and bacterial translocation. As a conclusion the authors again recall the strain specific actions of probiotics in different clinical situations and that so far probiotics play a role in rotaviral and post antibiotic diarrhoea and pouchitis. An important issue still to be solved in order to confidently recommend probiotics as efficacious therapy is the regulatory aspect of probiotics. [Abstract/Link to Full Text]

Thun R, Gajewski Z, Janett F
Castration in male pigs: techniques and animal welfare issues.
J Physiol Pharmacol. 2006 Nov;57 Suppl 8189-94.
Castration in male pigs is usually performed during the first weeks of life without prior anesthesia. This technique, however, is known to induce acute pain and stress and will therefore not be tolerated any longer by animal welfare organizations. Practical and animal-friendly alternatives to surgical castration are the production of entire male pigs, semen sexing or immunological castration. Fattening boars has the benefits of better feed efficiency, higher lean meat yield and increased animal welfare due to no pain and stress of castration. The most important disadvantage in raising entire male pigs is the incidence of boar taint ranging between 10 and 75%. To identify tainted carcasses an accurate and rapid on-line method for detection of odorous compounds is absolutely necessary. Sperm sexing through flow cytometry is the only commercially available method at the moment but speed of separation is too low for practical application. Active immunization of boars against gonadotropin-releasing-hormone (GnRH) at the end of the fattening period results in a significant reduction of testicular weight and androstenone production while the benefits of daily growth gain, meat quality as well as welfare remain the same as in entire males. In the present review more detailed information is given about the various techniques, especially the practical application of immunocastration on a large scale base. [Abstract/Link to Full Text]

Gajewski Z, Thun R, Faundez R, Pawli?ski B
The influence of alpha-adrenergic receptors stimulator and blockers and beta-blocker on the ovary and endocrinological activity in heifers during superovulation.
J Physiol Pharmacol. 2006 Nov;57 Suppl 8173-88.
Twenty five Holstein-Friesian heifers, clinically normal and with regular oestrous cycles, were used for induction of superovulation (PMSG-PGF(2)alpha-Neutra-PMSG). Animals were divided into 5 groups receiving: I - detomidine (40 microg/kg b.w.), II - doxazosin (0.2 mg/kg b.w.), III - yohimbine HCL 1% (1 ml/50 kg b.w.), IV - carazolol (0.01 mg/ kg b.w., i.v.), and V - physiological saline (1 ml/50 kg b.w.). The heifers with PGF2 alpha-induced cycles were treated with the substances 88 hrs after being given a single i.m. injection of 2500 IU PMSG. All animals were examined by ultrasonography, and by the number and size of ovarian follicles > 3 mm in diameter. The follicles were divided into 3 groups according to the diameter. Blood plasma was stored at -20 degrees C until LH, P4, E2 and PGFM analyses. In the control (V) group, two waves of follicle growth were observed. Yohimbine produced a significant blockage of ovulation. The mean number of corpora lutea in the group III was significantly lower than that in the control group (p< 0.02). No significant differences in the number of corpora lutea were observed between the groups I, II and III. The increase in E2 concentrations could be the response to the PMSG treatment with two waves of growth of large follicles before and after ovulation. Pulsatile LH release was altered by yohimbinum injection, however, the greater amplitude of pulses immediately following yohimbinum administration are suggestive of a positive influence of the alpha-2 adrenergic receptors antagonist. Yohimbinum administration did not affect plasma concentration of examined hormones. There was a difference between the plasma levels of LH after the doxazosin injection. Single injection of the stimulators and blockers of adrenergic receptors did not affect superovulatory response in terms of the numbers of CL, unruptured follicles and embryos recovered. The affectivity of artificial insemination was not significantly different between the control group and the detomidinum groups, while in the yohimbinum group it was significantly lower. [Abstract/Link to Full Text]

Sousa NM, Ayad A, Beckers JF, Gajewski Z
Pregnancy-associated glycoproteins (PAG) as pregnancy markers in the ruminants.
J Physiol Pharmacol. 2006 Nov;57 Suppl 8153-71.
In the last years, a polymorphic family of placenta-expressed proteins has been discovered in ruminant species and used for pregnancy diagnosis. Pregnancy diagnosis is an important part in reproduction management of ruminants. The pregnancy-associated glycoproteins (PAG) are synthesized in the mono- and binucleate cells of the ruminant's trophectoderm. Part of them is released into maternal blood circulation where they can be assayed by different RIA and ELISA systems. Due to large variety of expressed molecules and to large variations in the post-translational processing of the glycoproteins, different immuno-systems present different ability to quantify the PAG released in blood. Recent investigations showed that surprisingly the level of milk production in ruminants can modify the concentration of PAG circulating in blood. On the whole, the data show that the RIA methods are very precise for measuring PAG concentrations in the maternal blood and milk of the ruminants. Different studies clearly indicate that milk can be used for pregnancy diagnosis in small ruminants. The sensitivity and specificity of this method are very high. The results showed the possibility of the use PAG in milk and in blood as pregnancy test. It is especially helpful in the diagnosis of gestation and in detection of embryonic mortality as a non stressed method in the pregnancy management in the ruminants. [Abstract/Link to Full Text]

Ferreira-Dias G, Costa AS, Mateus L, Korzekwa A, Redmer DA, Skarzynski DJ
Proliferative processes within the equine corpus luteum may depend on paracrine progesterone actions.
J Physiol Pharmacol. 2006 Nov;57 Suppl 8139-51.
Soon after ovulation, the corpus luteum (CL) starts secreting progesterone (P(4)), a hormone necessary for implantation. The aim of the study was to evaluate whether P(4) exerts an autocrine/paracrine action on luteal angiogenic activity and P(4), prostaglandin E(2) (PGE(2)) and NO production in the mare. Corpora hemorrhagica (CH) and mid-luteal phase CL (MCL) were cultured with (i) no hormone (Control); (ii) P(4); (iii) a P(4) precursor - pregnenolone; or (iv) a P(4) antagonist - onapristone [10(-4) M;10(-5) M; all steroids]. NO production decreased in MCL, with respect to CH, when treated with P(4) [10(-4) M] and pregnenolone [10(-5) M]. PGE(2) increased from CH to MCL, and showed a tendency to rise in pregnenolone treated luteal tissues (10(-4) M; p=0.06). In the CH, P(4) decreased with pregnenolone [10(-4) M] compared to control, P(4) [10(-5) M], onapristone [10(-4) M;10(-5) M] and pregnenolone [10(-5) M](p<0.05). In the MCL, pregnenolone [10(-5) M] decreased (p<0.05) and P(4) tended to decrease (p=0.06) bovine aortic endothelial cell (BAEC) mitogenesis. Onapristone [10(-4) M] increased BAEC proliferation with respect to P(4) (p=0.01). Since there was no direct effect of treatments on BAEC, these data suggest that long-lasting effects of P(4) and its precursor may inhibit angiogenic factor(s) production by equine MCL, preparing for CL functional and structural regression. [Abstract/Link to Full Text]

Gajewski Z, Faundez R, Thun R, Pawli?ski B
Adrenergic stimulation and blocking of hormonal secretion activity of cultured cow granulosa cells.
J Physiol Pharmacol. 2006 Nov;57 Suppl 8125-37.
The aim of this study was to determine the influence of alpha- and beta-stimulators (alpha-stimulator: detomidinum HCl) as well as blockers (alpha1-blocker: doxazosin, alpha2-blocker: yohimbinum HCL, beta-blocker: carazolol) on bovine granulosa cells culture from preovulatory follicles. The cell culture was passed in TCM 199 medium with 10% FCS and antibiotics. Tested substances were added to the culture medium in different concentrations. The experiment began when at least 80% of the wells were covered (in four well culture dish of NUNCK-DK). The culture medium was collected every 24 h for hormone analysis. Hormone levels of T, E2, and P4 were determined. The culture was used up to 120 hours. Our results showed a decrease in P-4 secretion after detomidinum addition for all tested concentrations. A slight testosterone level increase was seen in the first 24 hours and then its concentration remained at a constant low level. A slight increase in 17-beta estradiol secretion was also observed. After yohimbinum addition, a statistically significant decrease of progesterone was observed for all concentrations tested. No significant changes were observed at other hormones levels when compared with the control. Doxazosin, when added into the culture medium, did not cause any statistically significant changes in hormone secretions. The addition of carazolol caused a significant decrease in progesterone secretion after culturing for 48 hours. Changes observed in other hormones levels did not differ statistically from the control. These results seem to support the hypothesis that drugs stimulating and blocking adrenergic receptors may play some role in ovarian steroidogenesis in cows. [Abstract/Link to Full Text]

Kotwica J, Ciuk MA, Joachimiak E, Rowinski S, Cymborowski B, Bebas P
Carbonic anhydrase activity in the vas deferens of the cotton leafworm - Spodoptera littoralis (Lepidoptera: Noctuidae) controlled by circadian clock.
J Physiol Pharmacol. 2006 Nov;57 Suppl 8107-23.
The male reproductive tract of Lepidoptera is an ideal model for the study of the physiological role of peripheral clocks in insects. The latter are significant in the generation and coordination of rhythmic phenomena which facilitate the initial stages of sperm capacitation. This process requires the maintenance of pH in the upper vas deferens (UVD) aided by, among others, H+-ATPase. Our aim was to determine the potential involvement of carbonic anhydrase (CA) in this process, an enzyme tasked with generating protons subsequently utilized by H+-ATPase to acidify the UVD milieu in S. littoralis, during the time when the lumen of this organ is filled with sperm. We attempted to answer the question whether CA activity can be controlled by the biological oscillator present in the male reproductive tract of the cotton leafworm. Using PAGE zymography, the presence of CA was demonstrated in the UVD wall, but not in the luminal fluid nor in the sperm. Using histochemistry, it was shown that CA is active in the UVD epithelium, and that this activity varies throughout the day and is most likely controlled by an endogenous biological clock. Conversely, the application of CA inhibitors, acetazolamide and sodium thiocyanate, in conjunction with an analysis of H+-ATPase activity in the acidification the UVD environment shows that CA most likely does not play a direct role in the regulation of the pH in this organ. [Abstract/Link to Full Text]

Nowakowska A, Caputa M, Rogalska J
Seasonal changes in cryoprotectants concentrations in Helix pomatia snails.
J Physiol Pharmacol. 2006 Nov;57 Suppl 893-105.
Terrestrial snails are often exposed to freezing. Therefore, we investigated seasonal shifts in hemolymph concentrations of cryoprotectants such as glycerol and glucose. We also investigated whether summer acclimation to cold and short-day photoperiod induced synthesis of cryoprotectants in Helix pomatia snails. Concentrations of the both cryoprotectants were elevated in winter and reduced in summer. These changes, however, were not correlated with shifts in liver glycogen content. Summer acclimation to cold (5 degrees C) and short-day photoperiod evoked a selective increase in glycerol concentration. In conclusion, glycerol may play a role in adaptation of the snails to winter cold and glucose is rather unlikely to provide the cryoprotection. [Abstract/Link to Full Text]

Wojciechowski MS, Jefimow M
Is torpor only an advantage? Effect of thermal environment on torpor use in the Siberian hamsters (Phodopus sungorus).
J Physiol Pharmacol. 2006 Nov;57 Suppl 883-92.
The aim of our study was determine torpor use in the Siberian hamsters (Phodopus sungorus) tested in a wide gradient of ambient temperatures (T(a)). Experiments were done on fed and food-deprived animals acclimated to winter-like and summer-like conditions. We found that neither fed nor unfed hamsters acclimated to winter-like or summer-like conditions selected low T(a)'s and entered torpor. Instead, food deprivation led to selection of higher T(a)'s and slight lowering of body temperature (T(b)), especially during the rest-phase of the day. Our calculations show that this strategy may lead to higher energy savings than torpor would. We argue that torpor use is not a fixed strategy but is determined primarily by a thermal conditions available in the environment. [Abstract/Link to Full Text]

Walentynowicz K, Szefer M, Wojtal B, Terlecki P, Wrotek S, Kozak W
Role of prostaglandins in heme-induced fever.
J Physiol Pharmacol. 2006 Nov;57 Suppl 873-82.
Brain stroke is often accompanied by a high fever, which is insensitive to a blockade with classic antipyretic drugs known to inhibit the synthesis of prostaglandin E(2) (PGE(2)), a proximal mediator of fever associated with infection. The molecular mechanism of fever associated with stroke is mostly unknown, and has not been thoroughly investigated. One characteristics of the stroke is an extravasation of the erythrocytes into the brain tissue followed by a release of hemoglobin and free heme. In the present study we have tested the hypothesis that free heme itself can induce fever after releasing into the brain. The study was conducted on Sprague Dawley rats instrumented with biotelemetry devices to monitor deep body temperature, and implanted with brain cannulae projected to the lateral ventricle. We demonstrate that heme-L-lysinate microinfused intraventricularly (icv) induces a dose-dependent fever lasting ca. 8 hours. Injection of heme-L-lysinate provoked a significant elevation of PGE(2) in the rat cerebro-spinal fluid collected 3 hours post-injection. The fever induced by heme-L-lysinate was blocked by an icv injection of anti- PGE(2) antibody. It was not affected, however, by intraperitoneal administration of indomethacin, a cyclooxygenase inhibitor. We conclude that heme-induced fever may underlie the stroke fever. [Abstract/Link to Full Text]

Stojek W, Borman A, Glac W, Baracz-J�?wik B, Witek B, Kamyczek M, Tokarski J
Stress-induced enhancement of activity of lymphocyte lysosomal enzymes in pigs of different stress-susceptibility.
J Physiol Pharmacol. 2006 Nov;57 Suppl 861-72.
To evaluate a possible mechanism of stress-induced lymphopenic effect we assessed the activity of lymphocyte lysosomal enzymes (LE) under immobilization. The effects of immobilization stress on LE (AP, acid phosphatase, cathepsin D and L, beta-N-acetyl-glucosamidase) activity in lymphocytes, number of lymphocytes and plasma cortisol (COR) level in the peripheral blood were examined in the cross-bred Pietrain pigs showing genotypic (presence or lack of RyR1 gene mutation) and phenotypic (reactivity to halothane) differences. It was found that immobilization stress evoked an increase in LE which was concomitant with lymphopenia and a rise of COR level. The most pronounced enhancement of LE, which may reflect a tendency to lymphocyte cytolysis, was found in the recessive homozygotes RyR1 (nn) phenotypically defined as stress/halothane susceptible as well as in the heterozygotes RyR1 (Nn) included in the group of stress/halothane resistant. Despite this individual variability the stress-induced increase in LE activity was present in all the animals. It seems that a possibility of destruction (lysis) of lymphocyte cells should not be excluded as one of the causes of stress lymphopenia. [Abstract/Link to Full Text]

Soszynski D
Molecular mechanism of emotional fever--the role of nitric oxide.
J Physiol Pharmacol. 2006 Nov;57 Suppl 851-9.
The purpose of these studies was to assess the involvement of nNOS and iNOS inhibitors on stress fever caused by exposure to an open field in freely moving biotelemetered rats. Vinyl-L-NIO (N(5) - (1-Imino-3-butenyl) - ornithine, a neural nitric oxide synthase (nNOS) inhibitor, and aminoguanidine, an inducible nitric oxide synthase (iNOS) inhibitor, were injected into the lateral ventricle (icv) at a dose of 5 microg and 10 microg, respectively, and then immediately exposed to open field for 30 min. After exposure to the open field, rats not treated with NOS inhibitors responded with a rapid rise in T(b) and it was accompanied with an increase of motor activity. Both inhibitors significantly suppressed the stress fever. vL-NIO did not influence stress-induced rise in locomotor activity as well as did not change T(b) in unstressed rats. Since aminoguanidine caused a transient fall in T(b) below the baseline in rats exposed or not to open field and because this inhibitor suppressed stress-induced rise in locomotor activity, we concluded that nNOS expression inside the brain is critically involved in the rise in T(b) due to exposure to psychological stress. [Abstract/Link to Full Text]

Kozak W, Wrotek S, Walentynowicz K
Hypoxia-induced sickness behaviour.
J Physiol Pharmacol. 2006 Nov;57 Suppl 835-50.
Sickness behaviour (SB) consists of the set of adaptive responses of the host to severe infections and inflammation. It includes, among others, the thermoregulatory responses such as regulated increase (fever) and/or decrease (anapyrexia) of body temperature (T(b)), decrease of motor activity (lethargy), and loss of appetite (hypophagia) resulting in a transient loss of body weight. It is thought that SB is partially induced by the immune-derived mediators such as cytokines and prostaglandins acting on the central nervous system. It has repeatedly been shown, on the other hand, that severe infections (pneumonia, tissue septicemia) can impair processes of the gases exchange both in the lungs and in distal tissues including brain, which may lead to hypoxia of the affected organs. Therefore, we have tested the hypothesis that hypoxia may also provoke SB. The study was conducted on freely moving biotelemetered mice kept at 28 degrees C ambient and 12/12 h light/dark cycle. We demonstrate that mice exposed for 7 days to hypoxia (11%O(2)) displayed all symptoms of SB. Interleukin-6 deficient mice (IL-6 KO) revealed reduced SB symptoms under hypoxic conditions. Recovery of the hypoxia-exposed mice to a normal rhythm in T(b), motor activity and feeding was unaffected by mepacrine, a phospholipase A(2) blocker. The recovery, however, was significantly impaired by indomethacin, a cyclooxygenase inhibitor. Exposure to hypoxemia resulted in significant elevation of plasma IL-6 in both untreated and treated with lipopolysaccharide (LPS) mice. It inhibited, however, a generation of blood prostaglandins (PGE(2)) in mice. Based on these data we conclude that IL-6 and accumulation of free arachidonic acid in biomembranes contribute to hypoxemia- induced SB. [Abstract/Link to Full Text]

Rogalska J, Caputa M, Wentowska K, Nowakowska A
Stress-induced behaviour in adult and old rats: effects of neonatal asphyxia, body temperature and chelation of iron.
J Physiol Pharmacol. 2006 Nov;57 Suppl 817-34.
Perinatal asphyxia in mammals leads to iron accumulation in the brain, which results in delayed neurobehavioural disturbances, including impaired learning and abnormal alertness over their entire life span. The aim of this investigation was to verify our hypothesis that newborn rats, showing reduced normal body temperature, are protected against neurotoxicity of the asphyxia up to senescence. Alertness was studied in adult and old male Wistar rats after exposure to critical neonatal anoxia: (i) at physiological neonatal body temperature of 33 degrees C, (ii) at body temperature elevated to 37 degrees C, or (iii) at body temperature elevated to 39 degrees C (the thermal conditions remained unchanged both during anoxia and for 2 h postanoxia). To elucidate the effect of iron-dependent postanoxic oxidative damage to the brain, half of the group (iii) was injected with deferoxamine, a chelator of iron. Postanoxic behavioural disturbances were recorded in open-field, elevated plus-maze, and sudden silence tests when the rats reached the age of 12 and 24 months. Open-field stress-induced motor activity was reduced in rats subjected to neonatal anoxia under hyperthermic conditions. In contrast, these rats were hyperactive in the plus-maze test. Both the plus-maze and sudden silence tests show reduced alertness of these rats to external stimuli signalling potential dangers. The behavioural disturbances were prevented by body temperature of 33 degrees C and by administration of deferoxamine. [Abstract/Link to Full Text]

Caputa M
Animal responses to extreme conditions: a lesson to biomedical research.
J Physiol Pharmacol. 2006 Nov;57 Suppl 87-15.
Various animals must cope with some specific extreme environmental conditions and, as a consequence, they developed extremely efficacious adaptive defence responses. The mechanisms of the specific defences are more clearly visible in some species than in humans. Therefore, animal models of the human defence mechanisms should be selected accordingly. The appropriate, well responding models may be regarded as "biological amplifiers". This review is focussed on examples of effective defence against: (i) parturitional asphyxia, which extends fertility of mammals; (ii) diving asphyxia, which extends access of food in aquatic birds and mammals; (iii) endotoxemia, which provides survival of the fittest in mammals showing top fertility; (iv) deep hypothermia, which enables hibernating mammals to arouse. Each of the defences needs close co-operation of the cardiovascular, respiratory and temperature regulatory systems. Underlined problems regarding a choice of appropriate experimental animal models should stimulate renaissance of comparative physiology. [Abstract/Link to Full Text]

Konturek SJ, Konturek PC, Brzozowski T
Melatonin in gastroprotection against stress-induced acute gastric lesions and in healing of chronic gastric ulcers.
J Physiol Pharmacol. 2006 Nov;57 Suppl 551-66.
The degree of gastric damage following to exposition of the mucosa to noxious agents depends upon a balance between the factors promoting this damage and those activating the natural defense mechanisms. Recent findings, presented in this review, provide evidence that melatonin prevents the formation of acute gastric lesions induced by stress and accelerates healing of chronic gastric ulcers due to increase in the activity of nitric oxide (NO) synthase (NOS)-NO and cyclooxygenase (COX)-prostaglandin E(2) (PGE(2)) systems resulting in the increase of mucosal blood flow and mucosal integrity. Melatonin is produced and released into the circulation by the pineal gland and, in many times larger amounts, by the gastrointestinal tract. Due to its anti-inflammatory and anti-oxidant properties, melatonin may be one of the most efficient protective factors preventing the development of acute gastric damage and accelerating healing of chronic gastric ulcers probably due to reduction in proinflammatory cytokine production, scavenging of the radical oxygen species and activation of COX-PG and NOS-NO systems as well as stimulating the afferent sensory nerves in the brain-gut axis. [Abstract/Link to Full Text]

Kapica M, Laubitz D, Puzio I, Jankowska A, Zabielski R
The ghrelin pentapeptide inhibits the secretion of pancreatic juice in rats.
J Physiol Pharmacol. 2006 Dec;57(4):691-700.
Ghrelin, a 28 amino acids polypeptide was recognized as an endogenous ligand for the growth hormone secretagogue receptor. It turned out that the entire sequence of ghrelin is not necessary for performing the above-mentioned functions. It was suggested that 5 residues (Gly-Ser-Ser(n-octanoyl)-Phe, pentaghrelin) constituted functionally active part of the full-length polypeptide. Ghrelin-28 was found to inhibit pancreatic enzyme output in rats, though the effect of pentaghrelin was not studied so far. The study aimed to determine the involvement of pentaghrelin in pancreatic juice secretion in anaesthetized rats. Male Wistar rats (220 +/- 20 g body weight, b. wt.) were anesthetized, the external jugular vein and common biliary-pancreatic duct were cannulated. Pentaghrelin boluses (i.v., 1.2, 12, and 50 nmol kg(-1) b. wt.) were injected every 30 min with or without CCK-8 infusion, duodenal mucosal CCK(1) receptor blockade with tarazepide, vagotomy and capsaicin pretreatment. Pentaghrelin boluses reduced the volume of pancreatic-biliary juice, protein and trypsin outputs both under basal and CCK-8-stimulated conditions in a dose-dependent manner. However, exogenous pentaghrelin failed to affect the pancreatic secretion in rats subjected to vagotomy, capsaicin deactivation of afferents or pretreatment with Tarazepide. In conclusion, pentaghrelin may control exocrine pancreas secretion by affecting duodenal neurohormonal mechanism(s) involving CCK and vagal nerves in rats. [Abstract/Link to Full Text]

Laubitz D, Jankowska A, Nieminuszczy J, Wrzesi?ski M, Jaworski A, Romanowicz K, Matyjek R, Grzesiuk E, Zebrowska T, Zabielski R
Pancreatic secretion differs according to the genotype of growing pigs.
J Physiol Pharmacol. 2006 Dec;57(4):677-89.
The objective of this study was to investigate the secretion of pancreatic enzymes and antibacterial activity in weaned pigs of three pure breeds, Pietrain, Duroc and Polish synthetic line 990, to look for eventual differences related to the genotype. Six male pigs of each breed, about 24 kg mean body weight, were equipped with chronic pancreatic duct catheters and duodenal cannulas to assess pure pancreatic juice, and jugular vein catheters for blood withdrawal. Pancreatic juice was collected before and after the morning feeding. Protein output and enzyme activities revealed two distinct profiles: strong manifestation of the prandial phase in Pietrain and line 990 pigs, and weak manifestation in Duroc. The antibacterial activity did not follow the enzyme kinetics, and it was the strongest in pancreatic juice from Pietrain pigs. Postprandial insulinaemia was reduced in the order of: line 990>Pietrain>Duroc. A slight (not significant) tendency towards a reduction of leptin after feeding in synthetic line 990 corresponded with elevated secretion of pancreatic enzymes and plasma insulin. The presented results suggest that the prandial secretion of pancreatic juice differs according to genotype, and the differences may be in part related to release of insulin. [Abstract/Link to Full Text]

Takeuchi K, Aihara E, Sasaki Y, Nomura Y, Ise F
Involvement of cyclooxygenase-1, prostaglandin E2 and EP1 receptors in acid-induced HCO3- secretion in stomach.
J Physiol Pharmacol. 2006 Dec;57(4):661-76.
We investigated the cyclooxygenase (COX) isoforms as well as prostaglandin E receptor EP subtypes responsible for acid-induced gastric HCO(3)(-) secretion in rats and EP receptor-knockout (-/-) mice. Under urethane anesthesia, a chambered stomach (in the presence of omeprazole) was perfused with saline, and HCO(3)(-) secretion was measured at pH 7.0 using a pH-stat method and by adding 2 mM HCl. Mucosal acidification was achieved by exposing the stomach for 10 min to 50 or 100 mM HCl. Acidification of the mucosa increased the secretion of HCO(3)(-) in the stomach of both rats and WT mice, in an indomethacin-inhibitable manner. The acid-induced gastric HCO(3)(-) secretion was inhibited by prior administration of indomethacin and SC-560 but not rofecoxib in rats and mice. Acidification increased the PGE(2) content of the rat stomach, and this response was significantly attenuated by indomethacin and SC-560 but not rofecoxib. This response was also attenuated by ONO-8711 (EP1 antagonist) but not AE3-208 (EP4 antagonist) in rats and disappeared in EP1 (-/-) but not EP3 (-/-) mice. PGE(2) increased gastric HCO(3)(-) secretion in both rats and WT mice, and this action was inhibited by ONO-8711 and disappeared in EP1 (-/-) but not EP3 (-/-) mice. These results support a mediator role for endogenous PGs in the gastric response induced by mucosal acidification and clearly indicate that the enzyme responsible for production of PGs in this process is COX-1. They further show that the presence of EP1 receptors is essential for the increase in the secretion of HCO(3)(-) in response to mucosal acidification in the stomach. [Abstract/Link to Full Text]

Olszanecki R, Gebska A, Korbut R
Production of prostacyclin and prostaglandin E2 in resting and IL-1beta-stimulated A549, HUVEC and hybrid EA.HY 926 cells.
J Physiol Pharmacol. 2006 Dec;57(4):649-60.
Production of arachidonic acid (AA) metabolites - prostacyclin (PGI(2)) in large vessels and prostaglandin E(2) (PGE(2)) in microcirculation is intrinsically involved in maintenance of vascular wall homeostasis. EA.hy 926 is a hybrid cell line, is derived by fusion of HUVEC with A549 cells. The aim of this study was to examine the production of prostacyclin and PGE2 in resting and IL-1beta-stimulated EA.ha 926 cells, in comparison with its progenitor cells. Non-stimulated EA.hy 926 cells has been found to produce much lower amounts of prostacyclin than resting HUVEC. Resting hybrid cells produced more PGE(2) than prostacyclin, despite they expressed high levels of COX-1 and PGI(2) synthase. On the contrary to HUVEC and A549, EA.hy 926 cells did not respond to IL-1beta with COX-2 induction and increase of prostaglandin production, however they did it in response to lysophosphatidylcholine (LPC). The characteristics of EA.hy 926 cells in terms of the pattern of prostanoid formation could facilitate studies on endothelial metabolism and role of these important lipid mediators. [Abstract/Link to Full Text]

Kosior-Korzecka U, Bobowiec R
Leptin effect on nitric oxide and GnRH-induced FSH secretion from ovine pituitary cells in vitro.
J Physiol Pharmacol. 2006 Dec;57(4):637-47.
The secretion of gonadotrophins from anterior pituitary cells can be modulated by leptin and signals originating from the immune system, among others, by nitric oxide (NO). There are some studies that have demonstrated a role for leptin and NO in the regulation of FSH in rodents, however, no similar data are available in regards to ewes. Therefore, the objective of the present study was to analyse the leptin effect on GnRH-induced FSH secretion from the ovine anterior pituitary cells in vitro. Additionally, the influence of leptin on NO release and its role in the GnRH and leptin-modulated secretion of FSH from pituitary gland of ewes was investigated. The obtained results show that the influence of leptin on FSH secretion is biphasic. Leptin in concentration 10(-8) and 10(-7) M/l significantly enhances, whereas 10(-6) and 10(-5) M/l of leptin suppresses FSH secretion from the pituitary cells in comparison to the control. The secretion of FSH and NO release under the influence of leptin are in very high positive correlation (r=0.77). The inhibition of NO synthesis with L-NAME., instead, disables leptin from the stimulation of FSH secretion. [Abstract/Link to Full Text]

Lipi?ska S, Zebrowska-Badalla A, Lipi?ska J
Oxytocin release after bleeding in rat: the role of sympathetic and renin-angiotensin system.
J Physiol Pharmacol. 2006 Dec;57(4):627-36.
The aim of this experiment was to compare the role of renin-angiotensin and sympathetic nervous system in post-haemorrhagic mechanism of oxytocin release. Oxytocin content in venous dialysates was determined by radioimmunoassay. In control rats the release of oxytocin into dialysates did not change during whole experiment. The injection of captopril induced 2-fold higher oxytocin release, but caused no change in oxytocin release after bleeding. Superior cervical ganglionectomy (SCGx) 20 days before, caused 5-fold higher increase in oxytocin release than in control group. Injection of captopril in rats after SCGx, did not decrease the high level of oxytocin in dialysate. However, bleeding increased oxytocin release and 1 hour after bleeding the highest - 14-fold increase, took place. In the contrary to 14-fold increase in oxytocin release in animals with superior cervical ganglia (SCG), bleeding after SCGx caused only 2-fold higher oxytocin release. When SCGx, bleeding and injection of captopril were done simultaneously, oxytocin release remained on the control concentration level. We assumed that blockade of renin angiotensin system and sympathetic dennervation prevent the increase in oxytocin release after bleeding. On basis of our present experiments, it can be assumed that, in posthaemorrhagic oxytocin release into the blood, sympathetic innervation derived from SCGx, as well as, renin-angiotensin system are involved. [Abstract/Link to Full Text]

Ryszawa N, Kawczy?ska-Dr�zdz A, Pryjma J, Czesnikiewicz-Guzik M, Adamek-Guzik T, Naruszewicz M, Korbut R, Guzik TJ
Effects of novel plant antioxidants on platelet superoxide production and aggregation in atherosclerosis.
J Physiol Pharmacol. 2006 Dec;57(4):611-26.
Superoxide anion is produced in human platelets predominantly by Nox2-dependent NADPH oxidases. In vitro experiments have shown that it might play a role in modulating platelet functions. The relationship between platelet superoxide production and aggregation remains poorly defined. Accordingly, we aimed to study superoxide production and aggregation in platelets from subjects with significant cardiovascular risk factors (hypertension, hypercholesterolemia, smoking and diabetes mellitus) and from control individuals. Moreover, we studied the effects of novel polyphenol-rich extracts of Aronia melanocarpa (chokeberry) berries on platelet function in vitro. Superoxide production was significantly increased in patients with cardiovascular risk profile when compared to controls, while platelet aggregation in response to either collagen or thrombin were borderline higher, and did not reach statistical significance. Interestingly, no relationship was observed between platelet aggregation ex vivo and platelet superoxide production in either of studied groups. No correlation was found between endothelial function (measured by FMD) and platelet aggregation ex vivo either. Polyphenol-rich extracts of A. melanocarpa berries caused a significant concentration dependent decrease in superoxide production only in patients with cardiovascular risk factors, while no effect was observed in the control group. A. melanocarpa extracts abolished the difference in superoxide production between risk factor patients and controls. A. melanocarpa extracts exerted significant concentration dependent anti-aggregatory effects in both studied groups, which indicated that these effects may be independent of it's ability to modulate superoxide production. The anti-aggregatory effects of chokeberry extracts were similar irrespective of aggregation inducing agent (collagen or thrombin). Moreover, they appear to be independent of platelet NO release as NOS inhibition by L-NAME did not lead to their abrogation. [Abstract/Link to Full Text]

Lewko B, Go?os M, Latawiec E, Angielski S, Stepinski J
Regulation of cGMP synthesis in cultured podocytes by vasoactive hormones.
J Physiol Pharmacol. 2006 Dec;57(4):599-610.
The podocytes are highly differentiated cells playing a key role in glomerular filtration. Vasoactive factors including angiotensin II (Ang II) and cyclic guanosine 5' monophosphate (cGMP) are synthesized by these cells upon stimulation as well as in the basal state. In this study we have tested whether angiotensin II affects the total synthesis of cGMP in primary culture of rat podocytes. The cells were stimulated with atrial natriuretic peptide (ANP) and/or a nitric oxide (NO) donor, S-nitroso-N-acetyl penicillamine (SNAP), in the absence or presence of Ang II. The cGMP synthesis was determined by radioimmunoassay (RIA). ANP or SNAP alone increased the cGMP synthesis in podocytes although the effects were not additive unless Ang II was present in the medium. Ang II suppressed the ANP-dependent cGMP synthesis whereas SNAP-dependent cGMP production remained unaffected. These effects were prevented by a non-specific antagonist of Ang II receptors (AT), saralasin. Adversely, PD123319, a specific inhibitor of AT2 receptors, augmented inhibition of ANP-dependent and enhanced the NO-dependent cGMP production. Probenecid, an inhibitor of cGMP extrusion from the cells, suppressed the cGMP generation by both ANP and SNAP. We conclude that cGMP synthesis in cultured podocytes is modulated by angiotensin II and that two adversely acting receptors, AT1 and AT2 are involved in this effect. Additionally, production of cGMP might be intrinsically inhibited by cGMP accumulating inside the cells. [Abstract/Link to Full Text]

Recent Articles in Polish Journal of Pharmacology and Pharmacy

Borowicz KK, Sekowski A, Drelewska E, Czuczwar SJ
Riluzole enhances the anti-seizure action of conventional antiepileptic drugs against pentetrazole-induced convulsions in mice.
Pol J Pharmacol. 2004 Mar-Apr;56(2):187-93.
Riluzole, a pre- and postsynaptic modulator of glutamate transmission, administered alone at doses of 5 and 10 mg/kg did not affect pentetrazole-evoked seizures in mice. However, it enhanced the anti-seizure action of valproate, phenobarbital, ethosuximide, although not that of clonazepam, in this model of experimental epilepsy. Keeping in mind that riluzole did not change plasma levels of antiepileptic drugs, a pharmacokinetic interaction, at least in terms of free plasma levels, does not seem probable. Regarding undesired effects, riluzole (5 mg/kg) and its combination with valproate did not produce any motor or long-term memory impairment. In contrast, the concomitant treatment of riluzole (5 mg/kg) with valproate (144 mg/kg), phenobarbital (4.9 mg/kg), or ethosuximide (90 mg/kg), resulted in a moderate motor deficit, but not long-term memory impairment in the tested mice. In conclusion, the results of the present study suggest that riluzole might occur effective as an additive drug in the treatment of myoclonic or absence epilepsy in humans. [Abstract/Link to Full Text]

Rog�z Z, Skuza G, Ku?mider M, W�jcikowski J, Kot M, Daniel WA
Synergistic effect of imipramine and amantadine in the forced swimming test in rats. Behavioral and pharmacokinetic studies.
Pol J Pharmacol. 2004 Mar-Apr;56(2):179-85.
Our previous studies demonstrated that joint administration of a tricyclic antidepressant drug, imipramine (IMI) with the uncompetitive antagonist of NMDA receptor, amantadine (AMA), produced stronger "antidepressant" effect in the forced swimming test (Porsolt's test) than the treatment with either of drugs given alone. Since it has been suggested that, in addition to their other functions, dopamine and alpha(1)-adrenergic receptors may play a role in behavioral response in the forced swimming test, in the present study we examined the effect of sulpiride (dopamine D(2/3) receptor antagonist) and prazosin (alpha(1)-adrenergic receptor antagonist) on the effect of AMA given alone or in combination with IMI in the forced swimming test in rats. We also measured the level of IMI and its metabolite, desipramine, in the rat plasma and brain, 1 h after the forced swimming test. Joint treatment with IMI (5 or 10 mg/kg) and AMA (20 mg/kg) produced stronger antidepressant-like effect than either of agents given alone. Sulpiride (10 mg/kg) or prazosin (1 mg/kg) (ineffective in the forced swimming test) inhibited an antidepressant-like effect induced by co-administration of IMI and AMA. The active behaviors in that test did not reflect an increase in general activity, since combined administration of IMI and AMA failed to enhance the locomotor activity of rats, measured in the open field test. Also sulpiride and prazosin did not decrease the exploratory activity induced by co-administration of IMI and AMA. The above result suggests that the dopamine D(2/3) and alpha(1)-adrenergic receptors may contribute to the mechanism of synergistic action of IMI and AMA in the forced swimming test in rats. The pharmacokinetic interaction can be excluded, since AMA did not change significantly the antidepressant level in the rat plasma and brain, measured 1 h after exposure to the forced swimming test. [Abstract/Link to Full Text]

Kowalska A
Genetic basis of neurodegeneration in familial Alzheimer's disease.
Pol J Pharmacol. 2004 Mar-Apr;56(2):171-8.
Alzheimer's disease (AD), the most common form of dementia, is characterized by two types of brain lesions, referred to as senile plaques and neurofibrillary tangles. Moreover, neuronal cell loss and synaptic degeneration appear in affected regions of the brain. A series of endoproteolytic cleavages of the amyloid precursor protein (APP) controlled by alpha, beta, and gamma-secretases leads to a formation of non-amyloidogenic (the alpha-secretase pathway) and amyloidogenic (the beta-secretase pathway) products which are essential for neurodegeneration. According to the "amyloid cascade hypothesis", the accumulation of amyloid beta (Abeta) peptides in the brain is a primary event in the pathogenesis of AD. One of the strong pieces of evidence supporting this hypothesis was the identification of pathogenic mutations within APP, presenilin 1 and presenilin 2 genes responsible for familial autosomal dominant AD. These mutations affect APP processing causing overproduction of Abeta42. Finding specific inhibitors of the Abeta42 generation is a major goal of AD drug development programs now and the key challenge for the treatment of the most devasting disease of human brain. [Abstract/Link to Full Text]

Reiter RJ, Tan DX, Gitto E, Sainz RM, Mayo JC, Leon J, Manchester LC, Vijayalaxmi E, Kilic U
Pharmacological utility of melatonin in reducing oxidative cellular and molecular damage.
Pol J Pharmacol. 2004 Mar-Apr;56(2):159-70.
This review briefly summarizes the actions of melatonin in reducing molecular damage caused by free radicals and associated oxygen- and nitrogen-based reactants. All the mechanisms by which melatonin is protective of such a wide variety of molecules, i.e. lipids, proteins, DNA, etc., and in such widely diverse areas of the cell and different organs are likely not yet all identified. Melatonin actions that have been identified include its ability to directly neutralize a number of toxic reactants and stimulate antioxidative enzymes. Furthermore, several metabolites that are formed when melatonin neutralizes damaging reactants are themselves scavengers suggesting that there is a cascade of reactions that greatly increase the efficacy of melatonin in styming oxidative mutilation. Suggested, but less well defined, processes which may contribute to melatonin's ability to reduce oxidative stress include stimulation of glutathione synthesis (an important antioxidant which is at high concentrations within cells), reducing electron leakage from the mitochondrial electron transport chain (which would reduce free radical generation), limiting cytokine production and inflammatory processes (actions that would also lower toxic reactant generation), and synergistic effects with other classical antioxidants (e.g. vitamins C, E and glutathione). Clearly which of these multiple mechanisms contribute to melatonin's high efficacy in curtailing oxidative damage remains to be clarified. Likewise, it is possible that the key action of melatonin in reducing molecular damage induced by oxygen and nitrogen-based metabolites remains to be identified. Finally, the review summarizes some of the large amount of data documenting the ability of melatonin to limit molecular and organ damage in two situations, i.e. ischemia-reperfusion and ionizing radiation, where free radicals are generally conceded as being responsible for much of the resulting tissue destruction. [Abstract/Link to Full Text]

Abstracts of the Twelfth Days of Neuropsychopharmacology. Ustron-Jaszowiec, May 26-28, 2003.
Pol J Pharmacol. 2003 Mar-Apr;55(2):275-314. [Abstract/Link to Full Text]

Kilic FS, Batu O, Sirmagul B, Yildirim E, Erol K
Intestinal absorption of digoxin and interaction with nimodipine in rats.
Pol J Pharmacol. 2004 Jan-Feb;56(1):137-41.
It is known that digoxin, which is a liposoluble cardiac glycoside, is well absorbed from intestine. In the present study, the absorption rates of digoxin from rat duodenum and the proximal and terminal parts of small intestine were determined in vitro. The isolated everted duodenum and intestinal sacs were put into oxygenated Tyrode solution at 37 degrees C. The Tyrode solution on the outer, mucosal side of intestinal segments contained 0.3 microM digoxin. Samples from the internal serosal side of the intestinal sacs were taken at 30, 60 and 120 min after the start of the experiments. The concentration of digoxin in the samples of fluid were determined using a radioimmunoassay method. The effect of nimodipine (0.1 and 0.2 mM) on digoxin absorption was also evaluated on the terminal segment of rat intestine. The interaction of nimodipine (0.5 mg/kg) and digoxin (0.2 mg/kg) was investigated in vivo when they were given perorally to rats. The duodenal absorption of digoxin was lower than in the small intestine. The highest absorption occurred in the terminal segment of the small intestine. Nimodipine increased the absorption of digoxin from the terminal segment of intestine in vitro, while it did not affect the serum digoxin concentration in vivo. [Abstract/Link to Full Text]

Gabryel B, ?abuzek K, Ma?ecki A, Herman ZS
Immunophilin ligands decrease release of pro-inflammatory cytokines (IL-1beta, TNF-alpha and IL-2 in rat astrocyte cultures exposed to simulated ischemia in vitro.
Pol J Pharmacol. 2004 Jan-Feb;56(1):129-36.
The aim of present study was to evaluate the effects of immunophilin ligands (cyclosporin A, FK506 and rapamycin) on the simulated ischemia-induced release of pro-inflammatory cytokines (IL-1beta, TNF-alpha and IL-2) in rat primary astrocyte cell cultures. Astrocytes were exposed to cyclosporin A (CsA) (0.25, 0.5, 1, 10, 20 and 50 microM), FK506 (1, 10, 100, 1000 nM) and rapamycin (10, 100, 500 and 1000 nM). In vitro simulated ischemia significantly increased secretion of IL-1beta, TNF-alpha and IL-2 by astrocyte cultures deprived of microglia (by shaking and incubating with L-leucine methyl ester). CsA (at concentrations of 10-50 microM), FK506 (at all used concentrations) and rapamycin (in dose-dependent manner) significantly attenuated IL-1beta release after 24 h exposure to ischemic conditions. Immunophilin ligands at all used concentrations significantly decreased TNF-alpha levels in culture media after 24 h exposure to ischemia. Moreover, significant decrease in IL-2 secretion at 0.25, 0.5, 1 and 50 microM CsA and FK506 at concentrations of 100 and 1000 nM were observed. The results suggest that immunophilin ligands may regulate glial activity during ischemia by affecting the release of pro-inflammatory cytokines. [Abstract/Link to Full Text]

Senthilkumar R, Nalini N
Glycine prevents hepatic fibrosis by preventing the accumulation of collagen in rats with alcoholic liver injury.
Pol J Pharmacol. 2004 Jan-Feb;56(1):121-8.
We studied the effect of administering glycine, a non-essential amino acid, on liver collagen content and its characteristics in experimental hepatotoxic Wistar rats. All the rats were fed standard pellet diet. Hepatotoxicity was induced by orally administering ethanol (7.9 g kg(-1)) for 30 days. Control rats were given isocaloric glucose solution. Glycine was administered subsequently at a dose of 0.6 g kg(-1) po every day, along with alcohol for the next 30 days. Alcohol administration significantly elevated the levels of liver hydroxyproline and total collagen content, cross-linked fluorescence, shrinkage temperature and lipid peroxidation, whereas it significantly decreased the solubility of liver collagen as compared with the control rats. Simultaneous glycine supplementation to alcohol-fed rats significantly reduced the levels of liver hydroxyproline and total collagen content, cross-linked fluorescence, shrinkage temperature and lipid peroxidation and enhanced the solubility of liver collagen as compared with the unsupplemented alcohol-fed rats. In conclusion, administration of glycine had a positive influence both on the quantitative and qualitative properties of hepatic collagen in alcoholic liver injury. [Abstract/Link to Full Text]

Tam�si V, Riedl Z, Dobozy O, Falus A, Vereczkey L, Monostory K
In vitro induction of cytochrome P450 enzymes in hepatocytes isolated from the regenerating rat liver.
Pol J Pharmacol. 2004 Jan-Feb;56(1):113-9.
Partial hepatectomy results in the loss of cytochrome P450 enzymes. During regeneration, the levels of cytochrome P450 activities, apoproteins and mRNA are reduced. Our present study investigated CYP1A, CYP2E1 and CYP3A induction in the cells of rat liver regenerating for 1, 3, 7, or 14 days. Hepatocytes were isolated from regenerating liver of hepatectomized rats and treated with enzyme inducers: 3-methylcholanthrene, imidazole and dexamethasone. CYP1A enzymes of the cells isolated from regenerating liver were inducible by 3-methylcholanthrene. The rate of induction of the cells from 3-day regenerating liver by 3-methylcholanthrene was three times higher than that of the hepatocytes of sham-operated rats. Dexamethasone caused about two- or three-fold stronger elevation of CYP3A in the cells of 1-, 3- and 7-day regenerating liver than in hepatocytes of sham-operated animals. However, the degree of CYP2E1 induction by imidazole was the same (about 2.5-fold) at each regenerating time as it was detected in the hepatocytes of sham-operated animals. In conclusion, the inducibility of the cells was retained at each regenerating time, but the degree of induction showed some differences. [Abstract/Link to Full Text]

Olakowska E, Olakowski M
Influence of a new propranolol analogue on the rabbit myocardium in vitro.
Pol J Pharmacol. 2004 Jan-Feb;56(1):105-11.
Experiments with electrophysiology of the heart have been essential for the progress in diagnostics and pharmacotherapy of cardiovascular diseases. The aim of the study was to establish the influence of a new propranolol analogue on mechanical and bioelectrical activity of the rabbit heart in vitro. In the experiment, propranolol (1-isopropylamino-3-[1-naphthoxy]-2-propanol hydrochloride) and its newly synthesized analogue (1-[1,1-dimethyl-ethyl-amino]-3-[1-naphthoxy]-2-propanol hydrochloride) were used. Atrial trabecules were cut from right rabbit atrium. Each preparation consisted of cardiomyocytes and sino-atrial node cells. Preparations were stimulated with square pulses of direct current at a voltage of 20V, rate of 2 Hz and 1 ms duration. Propranolol and its analogue were applied at gradded concentration 10(-2) M, 10(-3) M, 10(-4) M, 10(-5) M, 10(-6) M and 10(-7) M. Mechanical (force of contraction, time of contraction and relaxation) and bioelectrical (amplitude and duration of action potentials) activities were examined. Bioelectrical activity of preparations was recorded using intracellular microelectrodes. LD(50) for new analogue was determined. Analogue diminished force of contraction and shortened time of contraction and relaxation of myocardium and decreased amplitude and duration of action potentials in sino-atrial node cells. It influenced mechanical and bioelectrical parameters to lesser degree than propranolol. [Abstract/Link to Full Text]

Buczko W, Kubik A, Kucharewicz I, Chabielska E
Antithrombotic effect of captopril and enalapril in young rats.
Pol J Pharmacol. 2004 Jan-Feb;56(1):97-104.
Angiotensin converting enzyme inhibitors (ACE-Is) are the main drugs used in the treatment of essential hypertension and congestive heart failure in adults. Recently, we have demonstrated the antithrombotic effect of captopril (CAP) and enalapril (ENA) in venous thrombosis model in adult rats. One might also suggest the beneficial effect of those drugs on hemostasis in young individuals. Two months old male Wistar rats were used in the study. Acute administration of CAP at a dose of 50 and 100 mg kg(-1) significantly reduced the venous thrombus weight. Dose-dependent reduction in the thrombus weight was also observed in ENA (3, 10, 30 mg kg(-1))-treated rats. Strong reduction in the thrombus weight was also seen after chronic administration of CAP (2 x 25 mg kg(-1)) and ENA (1 x 15 mg kg(-1)). Both drugs given chronically reduced the frequency of thrombi. Systolic blood pressure was reduced to similar extent after acute and chronic application of the drugs. CAP shortened euglobulin clot lysis time (ECLT) when given acutely (100 mg kg(-1)) and chronically (2 x 25 mg kg(-1)). ENA decreased ECLT only when given at multiple doses (1 x 15 mg kg(-1)). None of the drugs changed prothrombin time or activated partial tromboplastin time. We conclude that CAP and ENA possess antithrombotic effect in young individuals. Activation of the fibrinolytic pathway seems to play an important role in the mechanism of their antithrombotic action. [Abstract/Link to Full Text]

Das N, Das Chowdhury T, Chattopadhyay A, Datta AG
Attenuation of oxidative stress-induced changes in thalassemic erythrocytes by vitamin E.
Pol J Pharmacol. 2004 Jan-Feb;56(1):85-96.
The oxidative stress status of the transfusion-dependent Ebeta- and beta-thalassemia patients were studied before and after treatment with vitamin E for a period of four weeks. The level of cellular vitamin antioxidants viz. ascorbic acid and vitamin E in the thalassemia patients were found to be considerably lower compared to normal subjects. The activities of enzymatic antioxidants viz. catalase, glutathione peroxidase and glutathione reductase were found to be drastically reduced in untreated Ebeta- and beta-thalassemic patients when compared to normal subjects. However, the activity of superoxide dis-mutase was found to be increased in both types of untreated thalassemic patients when compared to normal individuals. An increase in superoxide dismutase and a decrease in catalase activity reflects the presence of a severe oxidative stress situation in the erythrocytes of the untreated transfusion dependent Ebeta- and beta-thalassemia patients. Changes in erythrocyte membrane protein pattern in untreated Ebeta- and beta-thalassemia patients when compared to normal erythrocyte further confirm the presence of continued oxidative stress in the ailing thalassemic erythrocytes. All these changes in the antioxidant status as well as the changes in the erythrocyte membrane proteins are ameliorated to considerable extent when the transfusion-dependent Ebeta- and beta-thalassemia patients were treated with vitamin E at a dose of 10 mg/kg/day for a period of four weeks. The patients during the treatment period did not exhibit any side effects and gained in body weight indicating a healthy status. The present study reveals that the lipophilic antioxidant vitamin E could be useful in the management of transfusion-dependant Ebeta- and beta-thalassemia patients. [Abstract/Link to Full Text]

Manivasagam T, Subramanian P
Monosodium glutamate affects the temporal characteristics of biochemical variables in Wistar rats.
Pol J Pharmacol. 2004 Jan-Feb;56(1):79-84.
Monosodium glutamate (MSG) was administrated chronically for 60 days to Wistar rats and 24 h rhythms of glucose, cholesterol, total protein and alkaline phosphatase were studied. MSG treatment was found to cause acrophase delays in the glucose and alkaline phosphatase rhythms and advances in acrophases of cholesterol and total protein levels. Amplitude and mesor values of these rhythms were found to be altered during MSG treatment. Glutamate levels in the brain were found to be significantly increased, which could alter these biochemical rhythms by modulating the transmission in retinohypothalamic tract and in the hypothalamic nuclei, probably including suprachiasmatic nuclei. [Abstract/Link to Full Text]

Haider S, Saleem S, Shameem S, Ahmed SP, Parveen T, Haleem DJ
Is anorexia in thioacetamide-induced cirrhosis related to an altered brain serotonin concentration?
Pol J Pharmacol. 2004 Jan-Feb;56(1):73-8.
Anorexia or loss of appetite, one of the most typical symptoms observed in experimental and human cirrhosis, has been proposed to be associated with altered brain serotonin (5-HT) metabolism. In order to evaluate this hypothesis, brain 5-HT, its precursor tryptophan (TRP) and its metabolite 5-hydroxyindole-acetic acid (5-HIAA) were measured in brains of rats with thioacetamide (TAA)-induced liver cirrhosis. Thioacetamide at a dose of 500 mg/l in drinking water was administered for 6 weeks and during this period food intake was carefully measured in order to monitor the loss of appetite or decrease in food intake observed in cirrhosis. Concentrations of brain TRP, 5-HT and 5-HIAA were measured by HPLC with electrochemical detection. In TAA-treated rats, concentrations of 5-HT, TRP and 5-HIAA were increased in brain (44%, 33% and 36% of controls, p < 0.01). In plasma and liver of cirrhotic rats, TRP levels were increased (195% and 43%; p < 0.01). Plasma glucose and albumin levels were decreased (50%; p < 0.01 and 31%). Food intake, growth rate and locomotor activity of TAA-treated rats also decreased (73%, 22% and 73% of controls; p < 0.01). The results of this study show that brain 5-HT concentration in rats is increased in TAA-treated rats and it may, therefore, play an important role in the pathogenesis of anorexia associated with TAA-induced cirrhosis. [Abstract/Link to Full Text]

Parada-Turska J, Czuczwar M, Ki? J, Czuczwar P, Cioczek A, ?uszczki J, Czuczwar SJ
Allopurinol does not affect the anticonvulsant activity of carbamazepine and valproate in maximal electroshock-induced convulsions in mice.
Pol J Pharmacol. 2004 Jan-Feb;56(1):67-72.
Allopurinol, an inhibitor of xanthine oxidase, is indicated in the management of patients with elevated serum and urinary uric acid levels. It was also reported to be beneficial in patients with epilepsy when added to traditional antiepileptic drug. Here, we investigated the effect of allopurinol upon the electrical seizure threshold and its effect on the protective efficacy of common antiepileptic drugs, carbamazepine (CBZ) and valproate (VPA) against maximal electroshock (MES)-induced convulsions in mice. We found that allopurinol administered at doses of 5, 15 or 45 mg/kg, did not affect electrical seizure threshold. When administered acutely or for a prolonged period of time (5 times every 24 h), it did not affect anticonvulsant activity of CBZ and VPAin MES. Free plasma concentration of both anticonvulsants was not affected by allopurinol given at a dose of 45 mg/kg for 5 days. Thus, our results did not support suggestions that allopurinol can be beneficial as add-on drug in the management of epilepsy at least in patients treated with CBZ or VPA. [Abstract/Link to Full Text]

J�?wiak L, ?ukawski K, Czuczwar SJ, Sieklucka-Dziuba M
Competitive NMDA receptor antagonists and agonists: effects on spontaneous alternation in mice exposed to cerebral oligemia.
Pol J Pharmacol. 2004 Jan-Feb;56(1):59-66.
The purpose of the present study was to investigate the effects of competitive NMDA receptor antagonists,D,L-(E)-2-amino-4-methyl-5-phosphono-3-pentenoic acid (CGP 37849) and its ethyl ester (CGP 39551), or agonist, N-methyl-D-aspartate (NMDA) on spontaneous alternation in mice exposed to cerebral oligemia. Alternation behavior was evaluated in an Y-maze. Transient cerebral oligemic hypoxia was induced by bilateral clamping of carotid arteries (BCCA) for 30 min under pentobarbital anesthesia. In BCCA mice, CGP 37849 (5 mg/kg, ip) impaired spontaneous alternation when given 48 h or 7 days after surgery. CGP 39551 (5 mg/kg, ip) had no effect.NMDA (50 mg/kg, sc) improved performance of the task in BCCA mice when tested 48 h after surgery. These results suggest that cerebral oligemic hypoxia induced by BCCA leads to functional disturbances in the central nervous system, such as spontaneous alternation impairment and increased susceptibility to NMDA receptor-related drugs. Adverse potential of cerebral oligemia may limit a therapeutic use of NMDA receptor antagonists. [Abstract/Link to Full Text]

Kotli?ska J
Are glycineB sites involved in the development of morphine tolerance?
Pol J Pharmacol. 2004 Jan-Feb;56(1):51-7.
Numerous data have indicated that competitive and non-competitive N-methyl-D-aspartate (NMDA) receptor antagonists attenuate the development of tolerance to the analgesic effect of morphine. This study extends these findings on the effects of glycine(B) site antagonist, L-701.324. Tolerance to the analgesic effect of morphine was measured in hot-plate test in Wistar rats. For 9 days, animals were first injected with vehicle or glycine(B) receptor antagonist, L-701.324 (2.5 and 5 mg/kg, po). The non-competitive NMDA receptor antagonist, MK-801 (0.05 or 0.1 mg/kg, ip) was used as a reference compound. The injection of L-701.324, MK-801 or saline was followed, 20 min later, by the injection of morphine (10 mg/kg, sc). Hot-plate latencies were determined 20 min after the second injection on odd-numbered days. The results indicated that chronic administration of glycine(B) site antagonist, L-701.324 decreased the analgesic effect of morphine and they may suggest that this substance at both used doses increased the development of morphine tolerance, whereas non-competitive NMDA antagonist, MK-801 at the dose of 0.1 mg/kg potentiated the analgesic effect of morphine and attenuated the development of morphine tolerance. [Abstract/Link to Full Text]

Waszkielewicz A, Bojarski J
Gamma-hydrobutyric acid (GHB) and its chemical modifications: a review of the GHBergic system.
Pol J Pharmacol. 2004 Jan-Feb;56(1):43-9.
Gamma-hydroxybutyric acid (GHB) is a naturally occurring substance with function of an inhibitory neurotransmitter in the central nervous system in mammals. GHB can be used as a medicine in narcolepsy (Xyrem) and for general anesthesia (sodium oxybate). It is also a popular drug of abuse, causing coma, addiction and severe withdrawal syndrome, and, therefore, demanding thorough studies on the GHBergic system and expanded research on toxicology of this compound. The aim of this review is to present the proved and some suggested mechanisms of its action from pharmacological point of view, which may help to properly treat intoxication or other pathological states caused by GHB ingestion. Some new GHB derivatives studied for analogous action and their present use are also described. [Abstract/Link to Full Text]

Ku?ak W, Sobaniec W, Wojtal K, Czuczwar SJ
Calcium modulation in epilepsy.
Pol J Pharmacol. 2004 Jan-Feb;56(1):29-41.
The ideal antiepileptic drug (AED) should correct the aberrant pathophysiology of epileptogenesis without interfering with normal neurotransmission A new group of drugs with antiepileptic efficacy, without sedative properties, would be an exciting prospect. Theoretical considerations and results from experimental animal models of epilepsy have put forward the possibility that calcium (Ca2+) antagonists may form such a group. The initiation of epileptogenic activity in the neuron is thought to be connected with the phenomenon known as "intrinsic burst firing", which is activated by an inward Ca2+ current. Ca2+ is described as the primary mediator of "excitotoxic" neuronal damage. Both necrotic and apoptotic cell death is associated with Ca2+ entry into the cells during status epilepticus. The Ca2+ channel blockers depressed epileptic depolarizations of neurons. In this review, we present anticonvulsant effects of cinnarizine, flunarizine, nifedipine, nimodipine, nicardipine, amlodipine, isradipine, niguldipine, diltiazem, verapamil and dantrolene in animal models of seizures. Also, a detailed analysis of interactions between Ca2+ blockers and AEDs was performed. Clinical trials in intractable epilepsy support to a certain degree antiepileptic properties of Ca2+ antagonists. [Abstract/Link to Full Text]

Be?towski J, Jamroz A
Adrenomedullin--what do we know 10 years since its discovery?
Pol J Pharmacol. 2004 Jan-Feb;56(1):5-27.
Adrenomedullin (ADM) is a 52-amino acid peptide with structural homology to calcitonin gene-related peptide (CGRP) initially isolated from human pheochromocytoma. ADM is synthesized by many mammalian tissues including the adrenal medulla, endothelial and vascular smooth muscle cells, myocardium and central nervous system. ADM binds to plasma membrane receptors composed of calcitonin receptor-like receptor (CRLR), a member of serpentine receptor superfamily, and receptor activity modifying protein (RAMP) type 2 or 3. ADM has also some affinity for CGR(1) receptor composed of CRLR and RAMP1. ADM dilates blood vessels in both endothelium-dependent and independent manner and decreases systemic arterial pressure. Intrarenally administered ADM increases natriuresis by vascular and tubular mechanisms. In addition, ADM inhibits migration and proliferation of vascular smooth muscle cells and attenuates myocardial remodelling by inhibiting protein synthesis in cardiomyocytes and proliferation of cardiac fibroblasts. ADM is expressed in various tissues from early stage of embryogenesis and is also synthesized in placenta, uterus and fetal membranes. Plasma ADM level is increased in arterial hypertension, acute coronary syndromes, heart failure, renal diseases and septic shock, being involved in the pathophysiology of these disorders. Experimental ADM treatment is beneficial in arterial and pulmonary hypertension, heart failure, septic shock and ischemia/reperfusion injury. Proadrenomedullin N-terminal peptide (PAMP) is another product of ADM gene which is co-secreted by ADM-producing tissues, with some effects similar and some opposite to ADM. [Abstract/Link to Full Text]

Koci? I, Hirano Y, Hiraoka M
Early exposure to hypertonic solution strongly intensifies the effects of K+ channel opener, rilmakalim, in guinea pig ventricular myocytes.
Pol J Pharmacol. 2003 Nov-Dec;55(6):1159-62.
We report here that early exposure of guinea pig ventricular myocytes to hypertonic solution (approximately 400 mOsm compared to 280 mOsm in isotonic solution) increased the potency of rilmakalim to evoke ATP-sensitive K+ current 10 times (pD2 = 7.44 +/- 0.11 compared to pD2 = 6.49 +/- 0.18 in isotonic solution) without changing Emax, and observed effect was completely reversed by glibenclamide at 1 microM. [Abstract/Link to Full Text]

Basta-Kaim A, Budziszewska B, Jaworska-Feil L, Le?kiewicz M, Tetich M, Kubera M, Scharpe S, Laso? W
Opposite effects of clozapine and sulpiride on the lipopolysaccharide-induced inhibition of the GR-mediated gene transcription in fibroblast cells.
Pol J Pharmacol. 2003 Nov-Dec;55(6):1153-8.
Previously, we have found that some antipsychotic drugs are able to inhibit glucocorticoid receptor (GR)-mediated gene transcription. Since these drugs are known not only to inhibit hypothalamic-pituitary-adrenal (HPA) axis activity, but also to modulate the immunological system, the aim of the present study was to compare the effect of sulpiride and clozapine on GR function under basal culture conditions and during activation by lipopolysaccharide (LPS). The effect of clozapine and sulpiride alone and with LPS, the immune system activator, on glucocorticoid-mediated gene transcription was investigated in fibroblast cells, stably transfected with a mouse mammary tumor virus--chloramphenicol acetyltransferase plasmid (LMCAT cells). Treatment of the cells with clozapine (3-10 microM) for 2 days significantly and in concentration-dependent manner decreased the chloramphenicol acetyltransferase (CAT) activity, while sulpiride (1, 3, 5 and 10 microM) was without any effect. LPS (1 microg/ml) given alone inhibited the corticosterone-induced gene transcription by ca. 35%. Clozapine (3, 5 and 10 microM) inhibited the effect of LPS (1 microM), while sulpiride, which alone had no effect on GR function, enhanced LPS (1 microM) action. The obtained results indicate that inhibition of GR-mediated gene transcription by LPS and clozapine can be a mechanism by which these compounds blocked some effects induced by glucocorticoids. Opposite effect of clozapine and sulpiride on LPS action may result from their distinct effect on activity of some kinases involved in regulation of GR transcriptional function and may determine their utility in the treatment of schizophrenia with or without immune system activation. [Abstract/Link to Full Text]

Skuza G, Rog�z Z
Sigma1 receptor antagonists attenuate antidepressant-like effect induced by co-administration of 1,3 di-o-tolylguanidine (DTG) and memantine in the forced swimming test in rats.
Pol J Pharmacol. 2003 Nov-Dec;55(6):1149-52.
The obtained results show that DTG, the sigma1/sigma2 receptor agonist, exerts a synergistic effect with memantine, an uncompetitive NMDA receptor antagonist, in the forced swimming test in rats, and that progesterone and BD 1047, the sigma1 receptor antagonists, counteract this effect. The results suggest that the sigma1 receptor subtype may contribute to the behavioral response induced by combined administration of DTG and memantine in Porsolt's test in rats. [Abstract/Link to Full Text]

Nowak G, Siwek M, Dudek D, Zieba A, Pilc A
Effect of zinc supplementation on antidepressant therapy in unipolar depression: a preliminary placebo-controlled study.
Pol J Pharmacol. 2003 Nov-Dec;55(6):1143-7.
A growing body of evidence implicates a derangement of zinc homeostasis in mood disorders. In general, unipolar depression is connected with low blood zinc levels that are increased by effective antidepressant therapy. A placebo-controlled, double blind pilot study of zinc supplementation in antidepressant therapy was conducted in patients who fulfilled DSM IV criteria for major (unipolar) depression. Patients received zinc supplementation (6 patients; 25 mg of Zn2+ once daily) or placebo (8 patients) and were treated with standard antidepressant therapy (tricyclic antidepressants, selective serotonin reuptake inhibitors). Hamilton Depression Rating Scale (HDRS) and Beck Depression Inventory (BDI) were used to assess efficacy of antidepressant therapy, and patients' status was evaluated before the treatment and 2, 6 and 12 weeks after its commencement. Antidepressant treatment significantly reduced HDRS scores by the 2nd week of treatment in both groups, and lowered BDI scores at the 6th week in zinc-treated group. Zinc supplementation significantly reduced scores in both measures after 6- and 12-week supplementation when compared with placebo treatment. This preliminary study is the first demonstration of the benefit of zinc supplementation in antidepressant therapy. The mechanism(s) may be related to modulation of glutamatergic or immune systems by zinc ion. [Abstract/Link to Full Text]

Czerny B, Pawlik A, Juzyszyn Z, My?liwiec Z
Effect of tamoxifen on bone mineral density and blood lipids in ovariectomized rats.
Pol J Pharmacol. 2003 Nov-Dec;55(6):1137-42.
Tamoxifen is widely used in breast cancer therapy and in the treatment of all stages of breast cancer including chemoprevention in women at high risk of the disease. The most important aspect of tamoxifen therapy concerns its influence on bone tissue and lipid metabolism. The aim of the study was to evaluate the effect of tamoxifen on bone metabolism and blood cholesterol levels in ovariectomized rats. The study was performed on Wistar rats treated with tamoxifen at 2 and 4 mg/kg/24 h. Total serum cholesterol and low density cholesterol were significantly increased in ovariectomized rats (3.24 mmol/l and 2.06 mmol/l) in comparison with sham operated control (2.68 mmol/l and 1.44 mmol/l) (p < 0.05). Total serum cholesterol and low density cholesterol in tamoxifen-treated rats were significantly decreased in comparision with the values in both sham-operated and ovariectomized control. Bone mineral content (BMC) and bone mineral density (BMD) of femurs of ovariectomized rats (0.32 g and 0.081 g/cm2) decreased significantly compared to sham-operated controls (0.42 g and 0.098 g/cm2) (p < 0.05). Tamoxifen prevented the bone mass reduction induced by ovariectomy. The treatment with tamoxifen at doses of 2 mg/kg/24 h and 4 mg/kg/24 h significantly increased BMC and BMD in comparison with ovariectomized control. The results suggest a beneficial influence of tamoxifen on bone tissue and lipid metabolism. [Abstract/Link to Full Text]

Le?kiewicz M, Budziszewska B, Jaworska-Feil L, Kubera M, Basta-Kaim A, Laso? W
Inhibitory effect of some neuroactive steroids on cocaine-induced kindling in mice.
Pol J Pharmacol. 2003 Nov-Dec;55(6):1131-6.
Some neuroactive steroids which positively modulate GABAA receptor activity suppress cocaine-induced kindling but a possible involvement of other neurochemical mechanism in their antiepileptogenic effect remains to be elucidated. To this end, in the present study, we evaluated effects of allopregnanolone, a positive modulator of the GABAA receptor; its isomer without GABAergic activity--isopregnanolone and a negative-modulator of GABAergic transmission--dehydroepiandrosterone sulfate on cocaine-induced kindling in mice. Animals were pretreated daily with either vehicle or neuroactive steroid and then given cocaine (45 mg/kg) for 12 days. After a 14-day washout period in which drugs were not administered, the mice were challenged with the same 45 mg/kg dose of cocaine. Isopregnanolone (5 mg/kg) and dehydroepiandrosterone sulfate (20 mg/kg) administered daily with cocaine decreased number of mice exhibiting seizures. Allopregnanolone (5 mg/kg) also showed strong tendency to suppress cocaine kindling, however, its effect did not reach statistical significance. None of the neuroactive steroids had effect on acute cocaine (75 mg/kg ip)-induced clonic seizures. Further biochemical study showed that the veratridine- but not K+ -stimulated release of D-[3H]-aspartate in hippocampal slices was higher in cocaine-kindled mice than in the control group. Isopregnanolone (100 microM) significantly attenuated the veratridine-induced D-[3H]-aspartate release in hippocampi of cocaine-kindled group. These data indicate that positive modulation of the GABAA receptors is not a critical feature of neuroactive steroids that would determine their ability to prevent the cocaine-induced kindling. [Abstract/Link to Full Text]

Filip M, Papla I, Nowak E, Przegali?ski E
Blocking impact of clozapine on cocaine locomotor and sensitizing effects in rats.
Pol J Pharmacol. 2003 Nov-Dec;55(6):1125-30.
In the present study, we attempted to determine the effects of an atypical antipsychotic drug clozapine on the locomotor activation as well as sensitization to cocaine in male Wistar rats. When given acutely to rats, cocaine (10 mg/kg, ip) increased 4-fold the animals' locomotor activity. Repeated administration (1-5 days) of cocaine (10 mg/kg, ip) to rats significantly enhanced on day 10 the locomotor activation induced by its challenge dose given after 5-day withdrawal (sensitization). When given in combination with acute cocaine, clozapine (10 mg/kg, but not 2.5-5 mg/kg) attenuated the locomotor effects of the psychostimulant. In animals pretreated with clozapine (5-10 mg/kg, but not 2.5 mg/kg) before each of the 5 daily cocaine injections, a significant dose-dependent reduction of the locomotor response of animals to the challenge dose of cocaine (10 mg/kg) was observed on day 10. A decrease in that response was also found in animals treated repeatedly with cocaine (days 1-5) and challenged with the psychostimulant combined with clozapine (10 mg/kg, but not 2.5-5 mg/kg) on day 10. The obtained results indicate the ability of clozapine to reduce both acute and sensitizing locomotor responses to cocaine. These findings seem to be in line with recent clinical reports showing that clozapine may be useful in the treatment of cocaine abuse, even in schizophrenic patients. [Abstract/Link to Full Text]

Wiero?ska JM, Szewczyk B, Pa?ucha A, Bra?ski P, Smia?owska M
Involvement of CRF but not NPY in the anxiety regulation via NMDA receptors.
Pol J Pharmacol. 2003 Nov-Dec;55(6):1119-24.
The study attempts to evaluate whether neuropeptide Y (NPY) and corticotropin-releasing factor (CRF) are involved in anxiogenic and anxiolytic reactions induced by NMDA receptor ligands. The animals were given MK-801 (1 mg/kg, ip), a non-competitive NMDA-receptor antagonist, which acts as anxiolytic agent, or NMDA (15 mg/kg, ip), which has an anxiogenic effect. The anxiogenic or anxiolytic actions of these compounds were evaluated in the plus-maze test. The animals, which were given MK-801, were administered BIBO 3304 (130 ng/0.5 microl/site) intraamygdalarly and the animals which were given NMDA were administered alpha-helical CRF (500 ng/0.5 microl/site). BIBO 3304 did not attenuate MK-801-induced anxiolysis and alpha-helical CRF abolished NMDA-induced anxiogenesis. Our results show that anxiogenic effect of NMDA is mediated via CRF1 receptors and anxiolytic action of MK-801 is not dependent on Y1 receptors. [Abstract/Link to Full Text]

Zamani K, Faghihi K, Mehranjani MS
Synthesis of some new 2,5-disubstituted 1,3,4-thiadiazoles containing isomeric pyridyl as potent antimicrobial agents.
Pol J Pharmacol. 2003 Nov-Dec;55(6):1111-7.
Several new 2,5-disubstituted derivatives of 1,3,4-thiadiazoles containing isomeric pyridyl were obtained from cyclization of corresponding thiosemicarbazides under acidic conditions. The newly synthesized compounds were characterized using different methods of spectroscopy such as IR, 1H-NMR, 13C-NMR, MS and elemental analysis. The minimal inhibitory concentration (MIC) results of screening of some of the synthesized compounds were also reported. Most of the synthesized compounds have been found to be active against both Gram-positive and Gram-negative bacteria at less than 3.6 mg/ml. The compound (10b) is most active against all seventeen used gram-positive and gram-negative bacteria. [Abstract/Link to Full Text]

Aktan B, Taysi S, G�m�?tekin K, U��nc� H, Memi?o?ullari R, Save K, Bakan N
Effect of macrolide antibiotics on nitric oxide synthase and xanthine oxidase activities, and malondialdehyde level in erythrocyte of the guinea pigs with experimental otitis media with effusion.
Pol J Pharmacol. 2003 Nov-Dec;55(6):1105-10.
Although the long-term administrations of macrolide antibiotics are effective for diffuse panbronchiolitis, otitis media with effusion (OME), and some other diseases, their mechanism of action has not been fully understood. In order to elucidate the mechanisms of possible effects of macrolide antibiotics on activities of erythrocyte nitric oxide synthase (NOS), xanthine oxidase (XO), and malondialdehyde (MDA) levels in experimental OME, we aimed to evaluate the effect of macrolide antibiotics (erythromycin, azithromycin, roxithromycin, and clarithromycin) using an experimental guinea pig otitis media model. Erythrocyte NOS, XO activities, and MDA level were measured in all groups. Erythrocyte NOS activities were significantly higher in erythromycin-, azithromycin-, roxithromycin-, and clarithromycin-treated groups than in the experimental group. Erythrocyte XO activities were significantly lower in erythromycin-, azithromycin-, roxithromycin-, and clarithromycin-treated groups than in the control group. However, erythrocyte XO activities in experimental group were significantly higher than those of control group. Erythrocyte MDA levels were significantly lower in erythromycin-, azithromycin-, roxithromycin-, and clarithromycin-treated groups than those of the experimental group. The MDA levels in erythromycin- and roxithromycin-treated groups were significantly higher than those of azithromycin-treated group. The MDA levels in azithromycin-treated group were significantly lower than those of roxithromycin-treated group. In conclusion, the present study shows that the macrolide antibiotics (erythromycin, azithromycin, roxithromycin, and clarithromycin) increase NOS activity, decrease XO activity and MDA level, which is an important indicator of oxidative stress. [Abstract/Link to Full Text]

Recent Articles in Acta Pharmacologica Sinica

Chen GQ, Wang LS, Wu YL, Yu Y
Leukemia, an effective model for chemical biology and target therapy.
Acta Pharmacol Sin. 2007 Sep;28(9):1316-24.
The rapid rise of chemical biology aimed at studying signaling networks for basic cellular activities using specific, active small molecules as probes has greatly accelerated research on pathological mechanisms and target therapy of diseases. This research is especially important for malignant tumors such as leukemia, a heterogeneous group of hematopoietic malignancies that occurs worldwide. With the use of a chemical approach combined with genetic manipulation, great progress has been achieved over the past few decades on the biological, molecular and cytogenetic aspects of leukemia, and in its diagnosis and therapy. In particular, discoveries of the clinical effectiveness of all-trans retinoic acid and arsenic trioxide in the treatment of acute promyelocytic leukemia and the kinase inhibitors Imatinib and Dasatinib in the treatment of chronic myelogenous leukemia not only make target therapy of leukemia a reality, but also push mechanisms of leukemogenesis and leukemic cell activities forward. This review will outline advances in chemical biology that help our understanding of the molecular mechanisms of cell differentiation and apoptosis induction and target therapy of leukemia. [Abstract/Link to Full Text]

Sarkar FH, Li YW
Targeting multiple signal pathways by chemopreventive agents for cancer prevention and therapy.
Acta Pharmacol Sin. 2007 Sep;28(9):1305-15.
In recent years, growing interest has been focused on the field of cancer prevention. Cancer prevention by chemopreventive agents offers significant promise for reducing the incidence and mortality of cancer. Chemopreventive agents may exert their effects either by blocking or metabolizing carcinogens or by inhibiting tumor cell growth. Another important benefit of chemopreventive agents is their nontoxic nature. Therefore, chemopreventive agents have recently been used for cancer treatment in combination with chemotherapeutics or radiotherapy, uncovering a novel strategy for cancer therapy. This strategy opens a new avenue from cancer prevention to cancer treatment. In vitro and in vivo studies have demonstrated that chemopreventive agents could enhance the antitumor activity of chemotherapeutics, improving the treatment outcome. Growing evidence has shown that chemopreventive agents potentiate the efficacy of chemotherapy and radiotherapy through the regulation of multiple signaling pathways, including Akt, NF-kappaB, c-Myc, cyclooxygenase-2, apoptosis, and others, suggesting a multitargeted nature of chemopreventive agents. However, further in-depth mechanistic studies, in vivo animal experiments, and clinical trials are needed to investigate the effects of chemopreventive agents in combination treatment of cancer with conventional cancer therapies. More potent natural and synthetic chemopreventive agents are also needed to improve the efficacy of mechanism-based and targeted therapeutic strategies against cancer, which are likely to make a significant impact on saving lives. Here, we have briefly reviewed the role of chemopreventive agents in cancer prevention, but most importantly, we have reviewed how they could be useful for cancer therapy in combination with conventional therapies. [Abstract/Link to Full Text]

Howells LM, Moiseeva EP, Neal CP, Foreman BE, Andreadi CK, Sun YY, Hudson EA, Manson MM
Predicting the physiological relevance of in vitro cancer preventive activities of phytochemicals.
Acta Pharmacol Sin. 2007 Sep;28(9):1274-304.
There is growing interest in the ability of phytochemicals to prevent chronic diseases, such as cancer and heart disease. However, some of these agents have poor bioavailability and many of the in-depth studies into their mechanisms of action have been carried out in vitro using doses which are unachievable in humans. In order to optimize the design of chemopreventive treatment, it is important to determine which of the many reported mechanisms of action are clinically relevant. In this review we consider the physiologically achievable doses for a few of the best studied agents (indole-3-carbinol, diindolylmethane, curcumin, epigallocatechin-3-gallate and resveratrol) and summarize the data derived from studies using these low concentrations in cell culture. We then cite examples of in vitro effects which have been observed in vivo. Finally, the ability of agent combinations to act synergistically or antagonistically is considered. We conclude that each of the compounds shows an encouraging range of activities in vitro at concentrations which are likely to be physiologically relevant. There are also many examples of in vivo studies which validate in vitro observations. An important consideration is that combinations of agents can result in significant activity at concentrations where any single agent is inactive. Thus, for each of the compounds reviewed here, in vitro studies have provided useful insights into their mechanisms of action in humans. However, data are lacking on the full range of activities at low doses in vitro and the benefits or otherwise of combinations in vivo. [Abstract/Link to Full Text]

Davis CD, Milner JA
Biomarkers for diet and cancer prevention research: potentials and challenges.
Acta Pharmacol Sin. 2007 Sep;28(9):1262-73.
As cancer incidence is projected to increase for decades there is a need for effective preventive strategies. Fortunately, evidence continues to mount that altering dietary habits is an effective and cost-efficient approach for reducing cancer risk and for modifying the biological behavior of tumors. Predictive, validated and sensitive biomarkers, including those that reliably evaluate "intake" or exposure to a specific food or bioactive component, that assess one or more specific biological "effects" that are linked to cancer, and that effectively predict individual "susceptibility" as a function of nutrient-nutrient interactions and genetics, are fundamental to evaluating who will benefit most from dietary interventions. These biomarkers must be readily accessible, easily and reliably assayed, and predictive of a key process(es) involved in cancer. The response to a food is determined not only by the effective concentration of the bioactive food component(s) reaching the target tissue, but also by the amount of the target requiring modification. Thus, this threshold response to foods and their components will vary from individual to individual. The key to understanding a personalized response is a greater knowledge of nutrigenomics, proteomics and metabolomics. [Abstract/Link to Full Text]

Kong AN
Special issue on "molecular targets, biomarkers and animal models for anti-cancer pharmacological research: potentials and challenges from chemoprevention to chemotherapeutic treatment".
Acta Pharmacol Sin. 2007 Sep;28(9):1261. [Abstract/Link to Full Text]

Salimi M, Ghahremani MH, Naderi N, Amini M, Salimi E, Amanlou M, Abdi K, Salehi R, Shafiee A
Design, synthesis and pharmacological evaluation of 4-[2-alkylthio-5(4)-(4-substitutedphenyl)imidazole-4(5)yl]benzenesulfonamides as selective COX-2 inhibitors.
Acta Pharmacol Sin. 2007 Aug;28(8):1254-60.
AIM: To design and synthesize a series of benzenesulfonamide derivatives, 4-[2-alkylthio-5(4)-(4-substitutedphenyl)imidazole-4(5)-yl]benzenesulfonamides (4a-4j), which are intended to act as cyclooxygenase-2 (COX-2) inhibitors with good COX-2 inhibitor activity, and which will exert anti-inflammatory activities in vivo. METHODS: Benzenesulfonamide derivatives were designed and synthesized through multi-step chemical reactions. All the synthesized compounds were evaluated in an in vitro assay. The active compound 4a-4f was selected for further evaluation in a carrageenan-induced rat paw edema model. RESULTS: Docking studies showed that compound 4 bind into the primary binding site of COX-2 with the sulfonamide SO2NH2 moiety interacting with the secondary pocket amino acid residues. In the in vitro assay, compound 4 inhibited COX-2 with an inhibition concentration IC(50) value of 1.23-8 nmol/L, compared to celecoxib with IC(50) value of 1.5 nmol/L. Compound 4b and 4c had good potency and selectivity in comparison to the celecoxib. In the in vivo model, compound 4a-4f exhibited a moderate potency to inhibit 50% carrageenan-induced paw edema with value of 1.58-4.3 mg/kg. In the latter experiment, compound 4c was the most active compound. CONCLUSION: The anti-inflammatory effects obtained for compound 4a-4j could be due to the presence of fluorine or hydrogen substituents in the para position of the phenyl ring of these compounds. [Abstract/Link to Full Text]

Yoo HH, Kim NS, Im GJ, Kim DH
Pharmacokinetics and tissue distribution of a novel PDE5 inhibitor, SK-3530, in rats.
Acta Pharmacol Sin. 2007 Aug;28(8):1247-53.
AIM: To investigate the pharmacokinetic profile and tissue distribution of a novel phosphodiesterase type 5 inhibitor, 5-ethyl-2-{5-[4-(2-hydroxy-ethyl)-piperazine-1-sulfonyl]-2-propoxy-phenyl}-7-propyl-3,5-dihydro-pyrrolo(3,2-d)pyrimidin-4-one (SK-3530), in rats after administration of the (14)C-labeled compound. METHODS: The pharmacokinetic parameters of SK-3530 were measured based on the total radioactivity and parent SK-3530 concentration in rat plasma after intravenous and oral administration. The tissue distribution of total radioactivity after a single oral administration of [(14)C]SK-3530 at a dose of 40 mg/kg was assayed. The plasma protein binding rates of SK-3530 were assessed by in vitro and ex vivo assay. RESULTS: The total radioactivity profiles showed linear pharmacokinetics. The maximum plasma concentration and area under the curve of the parent SK3530 were 10%-20% compared to those of the total radioactivity. After the oral administration of [14C]SK-3530, the radioactivity was widely distributed in all tissues, and the tissue/plasma ratio of the radioactivity 1 h after administration was calculated as 0.5-2.6 with the exception of excretory organs. A relatively high penetration was shown in the adrenal glands, liver, and lung. In vitro and ex vivo plasma protein binding assay by ultrafiltration showed a considerably high binding rate of more than 97%. CONCLUSION: SK-3530 was relatively well absorbed in the gastrointestinal tract and showed linear pharmacokinetics over the investigated dose range. SK-3530 had low oral bioavailability due to a high, first-pass metabolism. [Abstract/Link to Full Text]

Zhang YF, Chen XY, Dai XJ, Zhang YN, Liu QZ, Yu HL, Zhong DF
Influence of omeprazole on pharmacokinetics of domperidone given as free base and maleate salt in healthy Chinese patients.
Acta Pharmacol Sin. 2007 Aug;28(8):1243-6.
AIM: To investigate the influence of omeprazole on the pharmacokinetics of domperidone given as free base and maleate salt. METHODS: An open, randomized, 2-period crossover study with a washout period of 7 d was conducted in 10 healthy Chinese, male patients. In each study period, the patients were administered a single oral dose of 10 mg domperidone as free base or maleate salt on d 1, 20 mg omeprazole twice daily on d 2 and 3, and once on d 4. A single dose of 10 mg domperidone as free base or maleate salt was taken at 4 h after administration of omeprazole on d 4. Plasma samples were collected on d 1 and 4 after the administration of domperidone, and the plasma concentrations of domperidone were determined by a sensitive liquid chromatography-tandem mass spectrometry method. RESULTS: For free-base domperidone, pretreatment with omeprazole resulted in a 16% decrease in maximum concentration (C(max)), compared with administration alone (P<0.05). However, for maleate salt, with the exception of an increase in t(1/2), no pharmacokinetic parameters were significantly changed. When the free base and maleate salt were administered alone, no differences were found in any parameters between the 2 formulations. In contrast, when they were administered in the presence of omeprazole, the C(max) of domperidone given as free base was lower (25.9%) than that given as maleate salt (P<0.05). CONCLUSION: Pretreatment of omeprazole does not affect the absorption of domperidone maleate, but leads to a moderately decreased rate of absorption of the free base. [Abstract/Link to Full Text]

Zhang F, Li YH, Fan MW, Jia R, Xu QA, Guo JH, Yu F, Tian QW
Enhanced efficacy of CTLA-4 fusion anti-caries DNA vaccines in gnotobiotic hamsters.
Acta Pharmacol Sin. 2007 Aug;28(8):1236-42.
AIM: To evaluate the comparative immunogenicity and protective efficacy of the cytotoxic T-lymphocyte-associated antigen 4 (CTLA-4) fusion anti-caries DNA vaccines pGJA-P/VAX1, pGJA-P, and non-fusion anti-caries DNA construct pGLUA-P in hamsters. In addition, the ability of CTLA-4 to target pGJA-P/VAX1-encoding antigen to dendritic cells was tested in vitro. METHODS: All DNA constructs contain genes encoding the A-P regions of a cell surface protein (PAc) and the glucan binding (GLU) domain of glucosyltransferases (GTFs) of cariogenic organism Streptococcus mutans. Human dendritic cells were mixed with the CTLA-4-Ig-GLU-A-P protein expressed by pGJA-P/VAX1-transfected cells and analyzed by flow cytometry. Gnotobiotic hamsters were immunized with anti-caries DNA vaccines by intramuscular injection or intranasal administration. Antibody responses to a representative antigen PAc were assayed by ELISA, and caries protection was evaluated by Keyes caries scores. RESULTS: A flow cytometric analysis demonstrated that CTLA-4-Ig-GLU-A-P protein was capable of binding to human dendritic cells. pGJA-P/VAX1 and pGJA-P induced significantly higher specific salivary and serum anti-PAc antibody responses than pGLUA-P. Significantly fewer caries lesions were also observed in hamsters immunized with pGJA-P/VAX1 and pGJA-P. There was no significant difference in the anti-PAc antibody level or caries scores between pGJA-P/VAX1 and pGJA-P-immunized groups. CONCLUSION: Antigen encoded by CTLA-4 fusion anti-caries DNA vaccine pGJA-P/VAX1 could specifically bind to human dendritic cells through the interaction of CTLA-4 and B7 molecules. Fusing antigen to CTLA-4 has been proven to greatly enhance the immunogenicity and protective efficacy of anti-caries DNA vaccines. [Abstract/Link to Full Text]

Lai TJ, Hsu SF, Li TM, Hsu HC, Lin JG, Hsu CJ, Chou MC, Lee MC, Yang SF, Fong YC
Alendronate inhibits cell invasion and MMP-2 secretion in human chondrosarcoma cell line.
Acta Pharmacol Sin. 2007 Aug;28(8):1231-5.
AIM: Chondrosarcoma is a malignant primary bone tumor that responds poorly to both chemotherapy and radiation therapy. The aim of the present study was to investigate the effect of alendronate, a bisphosphonate, on the invasion and migration of human chondrosarcoma cells (JJ012). METHODS: JJ012 cells were treated with alendronate of various concentrations up to 100 micromol/L for a specified period, and then gelatin zymography and matrigel invasion assay was performed to study the effects of alendronate on matrix metalloproteinase (MMP)-2 activity and the invasion ability of JJ012 cells, respectively. RESULTS: Our data showed that alendronate exerted a dose- and time-dependent inhibitory effect on the invasion and migration of JJ012 cells. Furthermore, gelatin zymography and RT-PCR showed that alendronate treatment decreased the activity and mRNA levels of MMP-2 in a concentration-dependent manner. CONCLUSION: Our findings suggest that alendronate may reduce MMP-2 secretion at the transcriptional and translational levels, and inhibit the invasion of chondrosarcoma cell. Therefore, alendronate may be a potential candidate for the systemic therapy of chondrosarcomas, as well as other malignant diseases. [Abstract/Link to Full Text]

Hu WG, Liu T, Xiong JX, Wang CY
Blockade of sonic hedgehog signal pathway enhances antiproliferative effect of EGFR inhibitor in pancreatic cancer cells.
Acta Pharmacol Sin. 2007 Aug;28(8):1224-30.
AIM: To investigate the expression of sonic hedgehog (SHH) and epidermal growth factor receptor (EGFR) signal molecules in pancreatic cancer cells, and to assess the inhibitory effects through the blockade of the SHH and EGFR signaling pathways by cyclopamine and Iressa, respectively. METHODS: The expression of SHH and EGFR in pancreatic cancer cell lines (PANC-1, SUIT-2, and ASPC-1) was detected by RT-PCR and Western blot analysis. After treatment with different concentrations of cyclopamine, alone or in combination with Iressa, the antiproliferative effect on pancreatic cancer cells was analyzed by methyl thiazolyl tetrazolium assays. A flow cytometry analysis was used to detect the cellular cycle distribution and apoptosis of pancreatic cancer cells. RESULTS: All of the 3 pancreatic cancer cell lines expressed SHH, Smoothened (SMO), and EGFR. Cyclopamine could downregulate the expression of EGFR in all cell lines. Cyclopamine or Iressa could induce a growth inhibitory effect in a dose-dependent manner. Moreover, the combined use of 2.5 micromol/L cyclopamine and 1 micromol/L Iressa induced an enhanced inhibitory effect and a greater apoptosis rate than any agent alone. The percentage of the cell population of the G0/G1 and sub-G1 phases was significantly increased along with the increasing dose of cyclopamine and/or Iressa. CONCLUSION: The blockade of the sonic hedgehog signal pathway enhances the antiproliferative effect of the EGFR inhibitor through the downregulation of its expression in pancreatic cancer cells. The simultaneous blockade of SHH and EGFR signaling represents possible targets of new treatment strategies for pancreatic carcinoma. [Abstract/Link to Full Text]

Hu S, Chen SM, Li XK, Qin R, Mei ZN
Antitumor effects of chi-shen extract from Salvia miltiorrhiza and Paeoniae radix on human hepatocellular carcinoma cells.
Acta Pharmacol Sin. 2007 Aug;28(8):1215-23.
AIM: To investigate the antihepatocellular carcinoma effects of chi-shen extract (CSE) from the water-soluble compounds of Salvia miltiorrhiza and Paeoniae radix. METHODS: The effect of CSE on the growth of HepG2 cells (hepatocellular carcinoma cell line) was studied by 3-(4,5)-2,5-diphenyltetrazolium bromide assay. Apoptosis were detected through acridine orange (AO) and ethylene dibromide (EB) staining and DNA fragmentation assay. The effect of CSE on the cell cycle of HepG2 cells was studied by the propidium iodide staining method. The activation of caspases-3, -8 and -9 was examined by immunoassay kits. The transcription of the Bcl-2 family and p53 was detected by RT-PCR. RESULTS: Our data revealed that CSE strongly induced HepG2 cell death in a dose- and time-dependent manner. CSE-induced cell death was considered to be apoptotic by observing the typical apoptotic morphological change by AO/EB staining and DNA fragmentation assay. The induction of HepG2 cell death was caused by an induction of apoptosis for the sub-G1 proportion increase, the downregulation of Bcl-2, the upregulation of Bax and p53, and the activation of the caspases-3 and -9 pathways. CONCLUSION: These results clearly demonstrated that CSE was able to inhibit the proliferation of HepG2 cells and cause apoptosis. Moreover, the anticancer effects of CSE were related to the Bcl-2 family pathway and the activation of caspases-3 and -9 in HepG2 cells. [Abstract/Link to Full Text]

Moon DO, Kim MO, Lee JD, Choi YH, Lee MK, Kim GY
Molecular mechanisms of ZD1839 (Iressa)-induced apoptosis in human leukemic U937 cells.
Acta Pharmacol Sin. 2007 Aug;28(8):1205-14.
AIM: To investigate the molecular mechanisms of ZD1839-induced apoptosis in human leukemic U937 cells. METHODS: The inhibition of human leukemic U937 cell growth was assessed by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphnyl-2H-tetrazolim bromide (MTT) assays, lactate dehydrogenase (LDH) release, and cell cycle distribution. The expression of anti- and pro-apoptotic proteins was detected by Western blot analysis. RESULTS: This study demonstrated that ZD1839 induced apoptosis in leukemic U937 cells by the downregulation of Bcl-2, caspase activation and subsequent apoptotic features. Cotreatment with ZD1839 and the caspase-3 inhibitor z-DEVD-fmk blocked apoptosis, indicating that caspase-3 activation is at least partially responsible for ZD1839-induced apoptosis. The ectopic expression of Bcl-2 attenuated caspase-3 activation, PARP cleavage, and subsequent indicators of apoptosis, including sub-G1 DNA content and LDH release. These results indicate that the downregulation of Bcl-2 plays a major role in the initiation of ZD1839-induced apoptosis, and that the activation of a caspase cascade is involved in the execution of apoptosis. Furthermore, ZD1839 treatment triggered the activation of p38 mitogen-activated protein kinase (MAPK) and the down-regulation of c-Jun-N-terminal kinase (JNK), extracellular signal-regulated kinase (ERK) and phosphatidyl inositol 3-kinase (PI3K)/Akt. The inhibition of the ERK and PI3K/Akt pathways also significantly increased cellular death. CONCLUSION: ZD1839 activated caspase-3 and the inhibited Bcl-2 in human leukemic U937 cells through the downregulation of the ERK and PI3K/Akt pathways. [Abstract/Link to Full Text]

Liu Y, Sun Y, Zhu T, Xie Y, Yu J, Sun WL, Ding GX, Hu G
11 beta-hydroxysteroid dehydrogenase type 1 promotes differentiation of 3T3-L1 preadipocyte.
Acta Pharmacol Sin. 2007 Aug;28(8):1198-204.
AIM: To investigate the relationship between 11 beta-hydroxysteroid dehydrogenase type 1 (11beta-HSD1), a potential link between obesity and type 2 diabetes, and preadipocyte differentiation. METHODS: Mouse 11beta-HSD1 siRNA plasmids were transfected into 3T3-L1 preadipocytes (a cell line derived from mouse Swiss3T3 cells that were isolated from mouse embryo), for examination of the effect of targeted 11beta-HSD1 inhibition on differentiation of 3T3-L1 cells. Differentiation was stimulated with 3-isobutyl-1-methyxanthine, insulin, and dexamethasone. The transcription level of the genes was detected by real-time PCR. RESULTS: Lipid accumulation was significantly inhibited in cells transfected with mouse 11beta-HSD1 siRNA compared with non-transfected 3T3-L1 cells. Fewer lipid droplets were detected in the transfected cells both prior to stimulation and after stimulation with differentiation-inducing reagents. The expression of adipocyte differentiation-associated markers such as lipoprotein lipase and fatty acid synthetase were downregulated in the transfected cells. Similarly, the expression of preadipocyte factor-1, an inhibitor of adipocyte differentiation, was downregulated upon stimulation of differentiation and had no changes in the transfected cells. CONCLUSION: 11 beta-HSD1 can promote preadipocyte differentiation. Based on this, we propose that 11 beta-HSD1 may be an important candidate mediator of obesity and obesity-induced insulin resistance. [Abstract/Link to Full Text]

An XF, Yu JY, Feng Y, Chen BY, Zhang SL
Role of hypothalamus nociceptin/orphanin FQ in pre-ovulatory luteinizing hormone surge of estrogen and progesterone-primed, ovariectomized rats.
Acta Pharmacol Sin. 2007 Aug;28(8):1189-97.
AIM: To investigate the role of hypothalamus nociceptin/orphanin FQ (OFQ) and its endogenous receptor, the opioid receptor-like1 receptor (ORL1 receptor) in the estrus cycle of female rats. METHOD: Radioimmunoassay was used to detect the effect of the intracerebroventricular (icv) administration of OFQ and/or the ORL1 receptor antagonist [Nphe1]Nociceptin(1-13)NH2, that is, NC13 on luteinizing hormone (LH) levels of estrogen- and progesterone (EBP)-primed, ovariectomized (OVX) rats (EBP-primed OVX rats). RT-PCR, Western blotting, and immunohistochemistry techniques were adopted to observe the changes of OFQ and the ORL1 receptor in the pre-optic area (POA) and the medial basal hypothalamus (MBH) of the estrus cycle of female rat. RESULTS: Pre-ovulatory LH surges in EBP-primed, OVX rats were significantly reduced by icv administration of 20 and 200 nmol OFQ (P<0.05), and the effect of 20 nmol OFQ could be abolished by pretreatment with 20 nmol NC13. The OFQ mRNA level in the POA on pro-estrus was lowered markedly compared to diestrus and estrus (P<0.05), while the mRNA and protein levels of the ORL1 receptor showed no significant changes in the POA and MBH across the estrus cycle. Meanwhile, the number of OFQ-immunoreactive neurons in the medial POA, ventromedial hypothalamus, and the arcuate nucleus on pro-estrus was significantly decreased compared to diestrus and estrus (P<0.05). CONCLUSION: The inhibitory effect of OFQ on the LH surge of EBP-primed, OVX rats and its downregulation in POA and MBH on pro-estrus suggests that it might play a negative modulatory role in the estrus cycle. [Abstract/Link to Full Text]

Huang Y, Chen J, Li G, Cheng TY, Jiang MH, Zhang SY, Lu J, Yan S, Fan WW, Lu DR
Reversal of hyperglycemia by protein transduction of NeuroD in vivo.
Acta Pharmacol Sin. 2007 Aug;28(8):1181-8.
AIM: To test whether the neurogenic differentiation (NeuroD) protein could alleviate symptoms of diabetes mellitus by its transduction activity in vivo. METHODS: Type 1 diabetes mellitus in mice was induced by ip (intraperitoneal) injection of streptozotocin (150 mg/kg). One group of diabetic mice were intravenously injected with the NeuroD-EGFP (Enhanced Green Fluorescent Protein) (5 mg/kg, n=6) and the other group with EGFP (5 mg/kg, n=5). After the transduction of NeuroD-EGFP, the distribution of the protein was examined by means of frozen section under fluorescent microscope observation. We conducted RT-PCR and Real-time quantitative PCR to measure the transcription levels of insulin mRNA. Immunohistochemistry was utilized to detect the insulin protein. Radioimmunoassay was conducted to determine the serum insulin levels. Blood glucose levels and body weights were regularly recorded after the protein administration. RESULTS: The NeuroD protein can be transduced into cells in vivo with a high efficiency of nearly 100%. Insulin mRNA was highly expressed in NeuroD-treated diabetic mice, 38-fold higher than that of control group (P<0.05). Immunohistochemistry revealed enteric insulin expression in the NeuroD-treated diabetic mice. The fasting serum insulin level of the NeuroD-EGFP group (n=6) was 337+/-39 pg/mL, significantly higher than that of the control diabetic mice (n=5) which was 84+/-23 pg/mL (P<0.01, t-test). Records of blood glucose level also displayed alleviation of hyperglycemia after NeuroD administration (P<0.01, t-test, n=6). CONCLUSION: In vivo-transduced NeuroD in the small intestine remained functionally active and could ameliorate the non-fasting glucose levels of streptozotocin-induced, diabetic mice by inducing enteric insulin expression. [Abstract/Link to Full Text]

Wang HH, Zhou HY, Chen CC, Zhang XL, Cheng G
Propofol attenuation of renal ischemia/reperfusion injury involves heme oxygenase-1.
Acta Pharmacol Sin. 2007 Aug;28(8):1175-80.
AIM: To examine the protective effect of propofol in renal ischemia/reperfusion (I/R) injury and the role of heme oxygenase-1 (HO-1) in this process. METHODS: Sprague-Dawley rats were randomly divided into 3 groups: (i) sham-operated group; (ii) I/R group; and (iii) propofol group. Bilateral renal warm ischemia for 45 min was performed. After 2, 6, and 24 h reperfusion, blood samples and kidneys were collected for assessment of renal injury, and HO-1 expressions were analyzed by immunohistochemical analysis, RT-PCR and Western blotting. RESULTS: Blood urea nitrogen and serum creatinine levels in the propofol group were significantly lower than that in the I/R group at 24 h after reperfusion. The mean histological score by Palleros standard showed that propofol significantly attenuated renal I/R injury after 6 h reperfusion. Propofol increased HO-1 mRNA and protein levels 2 h after reperfusion, whereas HO-1 expressions were present at exceedingly low levels in the I/R group and the sham-operated group at same time point. Propofol also markedly increased HO-1 mRNA and protein levels than I/R at 6 and 24 h after reperfusion. CONCLUSION: These results suggest that propofol mitigates renal I/R injury in rats. This protection may be partly through the induction of the HO-1 expression. [Abstract/Link to Full Text]

Wang K, Feng YL, Wen FQ, Chen XR, Ou XM, Xu D, Yang J, Deng ZP
Decreased expression of human aquaporin-5 correlated with mucus overproduction in airways of chronic obstructive pulmonary disease.
Acta Pharmacol Sin. 2007 Aug;28(8):1166-74.
AIM: To investigate the relationship between aquaporin-5 (AQP5) expression and mucus overproduction in the airways of Chinese patients with chronic obstructive pulmonary disease (COPD) and the correlation with pulmonary function change. METHODS: Bronchial tissues were obtained from fiberoptic bronchoscopy and bronchial biopsy in West China Hospital from April to July 2004. Twenty-five patients were diagnosed as COPD patients, and another 20 were diagnosed as the control patients. The expressions of AQP5, mucin 5AC (MUC5AC), and mucin in bronchial tissues were detected by semiquantitative RT-PCR, in situ hybridization, and immunohistochemical and alcian blue-periodic acid-Schiff (AB-PAS) staining, respectively. RESULTS: Compared with the control group, an attenuated expression of AQP5 was detected throughout the bronchial tissues from patients with COPD (P<0.01), but no difference existed in the lung tissues (P>0.05). Simultaneously, increased staining of MUC5AC and mucus in submucosal glands were noted (P<0.01, respectively). Smoking attenuated the expressions of AQP5 and increased the staining of MUC5AC and mucus in submucosal glands in the COPD groups (P<0.01), while there were no significant differences observed in the control group (P>0.05). The decreased expression of AQP5 mRNA was correlated with forced expiratory volume in 1 s (FEV1)/forced vital capacity (FVC) (r=0.60, P<0.01), FEV1% predicted value (r=0.60, P<0.01), maximal expiratory flow in 50% vital capacity (V50) % predicted value (r=0.55, P<0.01), and maximal expiratory flow in 25% vital capacity (V25) % predicted value (r=0.45, P<0.01). The decreased expression was negatively correlated with MUC5AC mRNA of the epithelium airways (r=-0.45, P<0.01) and the AB-PAS-stained area of submucosal glands (r=-0.61, P<0.01). The upregulation of MUC5AC mRNA correlated with the positively AB-PAS-stained area of submucosal glands and correlated negatively with FEV1/FVC (r=-0.53; P<0.01), FEV1% predicted value (r=-0.53; P<0.01), V50% predicted value (r=-0.48; P<0.01), and V25% predicted value (r=-0.43; P<0.01). CONCLUSION: The attenuated gene expression of AQP5 existed in the airways of Chinese COPD patients, which was complicated by mucus hypersecretion. The decreased expression of AQP5 mRNA may be related to the severity of airflow obstruction. [Abstract/Link to Full Text]

Zhong JH, Chen XP, Yun ML, Li WJ, Chen YF, Yao Z
Low-dose carvedilol reduces transmural heterogeneity of ventricular repolarization in congestive heart failure.
Acta Pharmacol Sin. 2007 Aug;28(8):1161-5.
AIM: To study the effects of carvedilol on the transmural heterogeneity of ventricular repolarization in rabbits with congestive heart failure (CHF). METHODS: Rabbits were randomly divided into 3 groups: control, CHF and carvedilol treated CHF group. Monophasic action potential duration (MAPD) in the 3 myocardial layers was simultaneously recorded. RESULTS: All the rabbits in the CHF group had signs of severe CHF. Compared with the control group, the mean blood pressure and cardiac output were significantly decreased, while peripheral resistance was significantly increased in the CHF group. This proved that the CHF model was successful created with adriamycin in this study. Compared to the control group, the ventricular fibrillation threshold (VFT) was remarkably decreased and all MAPD of the 3 myocardial layers were extended in rabbits with CHF. However, the extension of MAPD in the midmyocardium was more obvious. The transmural dispersion of repolarization (TDR) was significantly increased in CHF. Low-dose carvedilol (0.25 mg/kg, twice daily) had no effects on ventricular remodeling. Treatment with low-dose carvedilol significantly increased VFT. Although the MAPD of the 3 myocardial layers were further prolonged in the carvedilol treated CHF group, the prolongation of MAPD in the midmyocardium was shorter than those in the epicardium and endocardium. Treatment with low-dose carvedilol significantly decreased TDR in CHF. CONCLUSION: In the present study, the transmural heterogeneity of ventricular repolarization increased in the rabbits with CHF. Low-dose carvedilol decreased the transmural heterogeneity of ventricular repolarization in CHF, which may be related to its direct electrophysiological property rather than its effect on ventricular remodeling. [Abstract/Link to Full Text]

Yan D, Cheng LF, Song HY, Turdi S, Kerram P
Electrophysiological effects of haloperidol on isolated rabbit Purkinje fibers and guinea pigs papillary muscles under normal and simulated ischemia.
Acta Pharmacol Sin. 2007 Aug;28(8):1155-60.
AIM: Overdoses of haloperidol are associated with major ventricular arrhythmias, cardiac conduction block, and sudden death. The aim of this experiment was to study the effect of haloperidol on the action potentials in cardiac Purkinje fibers and papillary muscles under normal and simulated ischemia conditions in rabbits and guinea pigs. METHODS: Using the standard intracellular microelectrode technique, we examined the effects of haloperidol on the action potential parameters [action potential amplitude (APA), phase 0 maximum upstroke velocity (V(max)), action potential amplitude at 90% of repolarization (APD(90)), and effective refractory period (ERP)] in rabbit cardiac Purkinje fibers and guinea pig cardiac papillary cells, in which both tissues were under simulated ischemic conditions. RESULTS: Under ischemic conditions, different concentrations of haloperidol depressed APA and prolonged APD(90) in a concentration-dependent manner in rabbit Purkinje fibers. Haloperidol (3 micromol/L) significantly depressed APA and prolonged APD(90), and from 1 micromol/L, haloperidol showed significant depression on V(max); ERP was not significantly affected. In guinea pig cardiac papillary muscles, the thresholds of significant reduction in APA, V(max), EPR, and APD(90) were 10, 0.3, 1, and 1 mumol/L, respectively, for haloperidol. CONCLUSION: Compared with cardiac conductive tissues, papillary muscles were more sensitive to ischemic conditions. Under ischemia, haloperidol prolonged ERP and APD(90) in a concentration-dependent manner and precipitated the decrease in V(max) induced by ischemia. The shortening of ERP and APD(90) in papillary muscle action potentials may be inhibited by haloperidol. [Abstract/Link to Full Text]

Jiang QS, Huang XN, Yang GZ, Jiang XY, Zhou QX
Inhibitory effect of ginsenoside Rb1 on calcineurin signal pathway in cardiomyocyte hypertrophy induced by prostaglandin F2alpha.
Acta Pharmacol Sin. 2007 Aug;28(8):1149-54.
AIM: To examine the antihypertrophic effect of ginsenoside Rb1 (Rb1) induced by prostaglandin F2alpha(PGF2alpha) in vitro and to investigate the possible mechanisms involved in the calcineurin (CaN) signal transduction pathway. METHODS: The cardiomyocyte hypertrophy induced by PGF2alpha and the antihypertrophic effect of Rb1 were evaluated in primary culture by measuring the cell diameter, protein content, and atrial natriuretic peptide (ANP) mRNA expression. ANP and CaN mRNA expressions, CaN and its downstream effectors NFAT3 and GATA4 protein expressions, and the intracellular free Ca2+ concentration ([Ca2+]i) were assayed by RT-PCR, Western blot, and fluorescent determination using Fura 2/AM, respectively. RESULTS: PGF2alpha (100 nmol/L) significantly increased the cardiomyocyte diameter, protein content and [Ca2+]i, and promoted ANP, CaN mRNA, and CaN/NFAT3/GATA4 protein expressions, which were inhibited by either Rb1 in a concentration-dependent manner (50, 100, and 200 microg/mL) or L-arginine (1 mmol/L). NG-nitro-L-arginine-methyl ester, a nitric oxide synthase inhibitor, could abolish the effects of L-arginine, but failed to change the effects of Rb1 in the experiments above. CONCLUSION: The present data implicate that Rb1 attenuates cardiac hypertrophy, the underlying mechanism may be involved in the inhibition of the Ca2+-CaN signal transduction pathway. [Abstract/Link to Full Text]

Zha RP, Xu W, Wang WY, Dong L, Wang YP
Prevention of lipopolysaccharide-induced injury by 3,5-dicaffeoylquinic acid in endothelial cells.
Acta Pharmacol Sin. 2007 Aug;28(8):1143-8.
AIM: To investigate the effect of 3,5-dicaffeoylquinic acid (3,5-diCQA) on lipopolysaccharide (LPS)-induced injury in human dermal microvascular endothelial cells (HMEC-1). METHODS: The anti-oxidant effect was detected using the malondialdehyde (MDA) assay in a rat liver microsome model of lipid peroxidation. Cell viability was analyzed using the 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide assay. Cell lipid peroxide injury was measured by lactate dehydrogenase (LDH) release. Apoptotic cells were detected by flow cytometry, and confirmed by DNA fragmentation analysis. Caspase-3 activity was measured using a specific assay kit. The level of intracellular reactive oxygen species (ROS) was determined by flow cytometry with a 2,7-dichlorodihydro-fluorescein diacetate fluorescence probe. RESULTS: The exposure of microsomes to ascorbate-Fe2+ resulted in lipoperoxidation according to an increase in the level of MDA. MDA formation decreased in a dose-dependent manner on treatment with 5, 10, or 50 micromol/L 3,5-diCQA. Treatment with LPS for 16 h resulted in a 60% decrease in cell viability and an increase in LDH release from 47.6% to 61.5%. DNA laddering was observed by agarose gel electrophoresis. The level of apoptotic cells peaked at 27% after treatment with LPS for 12 h. Following treatment with LPS for 12 h, intracellular ROS and caspase-3 activity increased. Pretreatment with 3,5-diCQA at 5, 10, or 50 micromol/L for 1 h attenuated LPS-mediated endothelial cell injury. The anti-apoptotic action of 3,5-diCQA was partially dependent on its capacity for anti-oxidation and the suppression of caspase-3 activity. CONCLUSION: 3,5-diCQA displays anti-oxidative and anti-apoptotic activity in HMEC-1 due to scavenging of intracellular ROS induced by LPS, and the suppression of caspase-3 activity. [Abstract/Link to Full Text]

Jia EZ, Yang ZJ, Zhu TB, Wang LS, Chen B, Cao KJ, Huang J, Ma WZ
Serum sodium concentration is significantly associated with the angiographic characteristics of coronary atherosclerosis.
Acta Pharmacol Sin. 2007 Aug;28(8):1136-42.
AIM: To explore the relationship between serum sodium concentration and coronary atherosclerosis. METHODS: The study population consisted of 896 consecutive patients (684 males and 212 females) who underwent coronary angiography for suspected or known coronary atherosclerosis. Smoking and drinking were investigated. The anthropometric measurements, including body mass index, systolic blood pressure and diastolic blood pressure, and the serum measurements, including sodium, potassium, chlorine, lipids, blood glucose, urea, creatinine, and uric acid for every patient were conducted. The severity of coronary atherosclerosis was defined by the Gensini score system. The statistical methods, including one-way ANOVA, Kruskal-Wallis test, Spearman correlation analysis, partial correlation analysis, multivariate stepwise linear regression analysis, and multinomial logistic regression analysis were employed to explore the relationship between serum sodium concentration and the Gensini score. RESULTS: The analysis of the Kruskal-Wallis test indicated that the distribution of the Gensini score (P=0.000) differed among the groups according to serum sodium concentration, quartile values of which were used as cut-off points. The Spearman correlation and partial correlation analysis controlling for gender, smoking status, and drinking status indicated that the Gensini score significantly correlated with the sodium concentration (r=-0.241, P=0.000 for the Spearman correlation, r=-0.114, P=0.000 for the partial correlation). The results from the multivariate stepwise linear regression analysis showed that the left ventricular ejection fraction (beta=-0.228, P=0.000), age (beta=0.137, P=0.010), glucose level (beta=0.129, P=0.000), and sodium level (beta=-0.106, P=0.004) were significantly and independently associated with the Gensini score. The results of the multinomial logistic regression analysis suggested that the hyponatremia was the risk factor for the higher Gensini score. CONCLUSION: The serum sodium concentration was significantly and negatively associated with the Gensini score; and the actual mechanism underlying the association needs further study. [Abstract/Link to Full Text]

Yue Y, Hu L, Tian QJ, Jiang JM, Dong YL, Jin ZY, Cheng YH, Hong X, Ge QS, Zuo PP
Effects of long-term, low-dose sex hormone replacement therapy on hippocampus and cognition of postmenopausal women of different apoE genotypes.
Acta Pharmacol Sin. 2007 Aug;28(8):1129-35.
AIM: To study the effects of long-term, low-dose sex hormone replacement therapy (HRT) on the volume and biochemical changes of the hippocampus in postmenopausal women carrying apolipoprotein E (apoE) gene epsilon3 or epsilon4. METHODS: Eighty-three postmenopausal women who had used a low dose of HRT for over 4 years were selected as the HRT group, and 99 postmenopausal women with matched age and education were enrolled as the control group. ApoE alleles were analyzed by PCR. Magnetic resonance imaging was performed to determine the volume of the brain hippocampus. Proton magnetic resonance spectroscopy was used to detect the biochemical changes in the anterior cingulate cortex and hippocampus in apoE epsilon4 and epsilon3 carriers. Six common cognitive tests were used to make an overall evaluation of cognitive function. RESULTS: Analysis with the apoE epsilon4 carriers showed that the volume of the hippocampus of the control group were significantly lower than those of the HRT group. The biochemical analysis showed that there was an increase of N-acetylaspartate (NAA)/total creatine (tCr) and a decrease of myoinositol (mI)/tCr in the hippocampus of apoE epsilon4 carriers in the HRT group, compared with the control group. For the apoE epsilon3 carriers, the least squares means (LSMEAN) of the HRT group was higher than that of the control group. CONCLUSION: This study showed that long-term, low dose HRT might be beneficial for reducing the risk of AD development in vulnerable postmenopausal women. Meanwhile, HRT could increase the LSMEAN of apoE epsilon3 carriers. [Abstract/Link to Full Text]

Wu XJ, Zheng YJ, Cui YY, Zhu L, Lu Y, Chen HZ
Propofol attenuates oxidative stress-induced PC12 cell injury via p38 MAP kinase dependent pathway.
Acta Pharmacol Sin. 2007 Aug;28(8):1123-8.
AIM: To investigate the neuroprotective effect of propofol and its intracellular mechanism on neurons in vitro. METHODS: Cell viability was determined with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide reduction. Apoptotic cell death was determined by Hoechst 33258 staining and a fluorescence-activated cell sorter. The caspase-3 activity was measured by fluorometric assay. Mitogen-activated protein (MAP) kinase phosphorylation was detected with Western blotting. RESULTS: The pretreatment of rat pheochromocytoma cell line PC12 with propofol (1-10 micromol/L) resulted in a significant recovery from hydrogen peroxide (H2O2)-induced cell death and the inhibition of H2O2 induced caspase-3 activation and PC12 cell apoptosis. Propofol inhibited the H2O2-induced p38 MAP kinase, but not c-Jun N-terminal kinase or extracellular signal-regulated kinase 1 and 2 activations. CONCLUSION: Propofol might attenuate H2O2-induced PC12 cell death through the inhibition of signaling pathways mediated by the p38 MAP kinase. [Abstract/Link to Full Text]

Tang C, Wu AH, Xue HL, Wang YJ
Tanshinone IIA inhibits endothelin-1 production in TNF-alpha-induced brain microvascular endothelial cells through suppression of endothelin-converting enzyme-1 synthesis.
Acta Pharmacol Sin. 2007 Aug;28(8):1116-22.
AIM: To investigate the effects of tanshinone IIA (Tan IIA) on the regulation of the production of endothelin (ET)-1 (including large ET-1), mRNA levels of ET-1, endothelin-converting enzyme-1 (ECE-1), endothelin-A receptor (ETA) and endothelin-B receptor (ETB) induced by TNF-alpha in rat brain microvascular endothelial cells (BMVEC). METHODS: The ET-1 release (including large ET-1) into the culture medium was determined by enzyme immunoassay. The levels of ET-1, ECE-1, ETA, and ETB mRNA were measured by RT-PCR. Endothelin receptor binding was also tested. RESULTS: The induction of ET-1 release by TNF-alpha from cultured BMVEC was dose-dependently reduced by Tan IIA, but large ET-1 levels progressively increased in response to Tan IIA; the mRNA expression of ET-1 was unaffected. Tan IIA also caused a decrease in ETA receptor mRNA and ECE-1 expression in a dose-dependent manner. Endothelin receptor binding was unaltered in BMVEC stimulated with TNF-alpha alone or a combination of TNF-alpha and Tan IIA. CONCLUSION: These findings suggest that Tan IIA may inhibit ET-1 production in TNF-alpha-induced BMVEC through the suppression of ECE-1 synthesis. [Abstract/Link to Full Text]

Liu C, Liu JK, Kan MJ, Gao L, Fu HY, Zhou H, Hong M
Morphine enhances purine nucleotide catabolism in vivo and in vitro.
Acta Pharmacol Sin. 2007 Aug;28(8):1105-15.
AIM: To investigate the effect and mechanism of morphine on purine nucleotide catabolism. METHODS: The rat model of morphine dependence and withdrawal and rat C6 glioma cells in culture were used. Concentrations of uric acid in the plasma were measured by the uricase-rap method, adenosine deaminase (ADA) and xanthine oxidase (XO) in the plasma and tissues were measured by the ADA and XO test kit. RT-PCR and RT-PCR-Southern blotting were used to examine the relative amount of ADA and XO gene transcripts in tissues and C6 cells. RESULTS: (i) the concentration of plasma uric acid in the morphine-administered group was significantly higher (P<0.05) than the control group; (ii) during morphine administration and withdrawal periods, the ADA and XO concentrations in the plasma increased significantly (P<0.05); (iii) the amount of ADA and XO in the parietal lobe, liver, small intestine, and skeletal muscles of the morphine-administered groups increased, while the level of ADA and XO in those tissues of the withdrawal groups decreased; (iv) the transcripts of the ADA and XO genes in the parietal lobe, liver, small intestine, and skeletal muscles were higher in the morphine-administered group. The expression of the ADA and XO genes in those tissues returned to the control level during morphine withdrawal, with the exception of the skeletal muscles; and (v) the upregulation of the expression of the ADA and XO genes induced by morphine treatment could be reversed by naloxone. CONCLUSION: The effects of morphine on purine nucleotide metabolism might be an important, new biochemical pharmacological mechanism of morphine action. [Abstract/Link to Full Text]

Xu XH, Hua YN, Zhang HL, Wu JC, Miao YZ, Han R, Gu ZL, Qin ZH
Greater stress protein expression enhanced by combined prostaglandin A1 and lithium in a rat model of focal ischemia.
Acta Pharmacol Sin. 2007 Aug;28(8):1097-104.
AIM: To investigate the effects of lithium (Li) and prostaglandin A1 (PGA1) on the expression of heat shock factor 1 (HSF-1), heat shock proteins (HSP), and apoptosis protease activating factor-1 (Apaf-1) induced by permanent focal ischemia in rats. METHODS: The rats were pretreated with a subcutaneous (sc) injection of Li for 2 d or a single intracerebral ventricle (icv) administration of PGA1 for 15 min before ischemic insult, or a combination of Li (sc, 1 mEq/kg, 2 d) and PGA1 (icv, 15 min prior to ischemic insult). Brain ischemia was induced by the permanent middle cerebral artery occlusion (pMCAO). Twenty-four hours after the occlusion, the expression of HSF-1, HSP, and Apaf-1 in the ischemic striatum were examined with Western blot analysis. RESULTS: The expression of HSF-1, heme oxygenase-1 (HO-1), HSP90alpha, and Apaf-1 were significantly increased, but the expression of HSP90beta was significantly decreased 24 h after the pMCAO. PGA1 and Li and their combination significantly enhanced the ischemia-induced elevation in the levels of HSF-1, HO-1, and HSP90alpha, and recovered HSP90beta expression, but decreased Apaf-1 levels in the ischemic striatum. CONCLUSION: The present study demonstrates that PGA1 and Li have synergistic effects on the enhancement of the expression of HSP, suggesting that the synergistic effects of PGA1 and Li in the rat model of permanent focal cerebral ischemia may be mediated by the enhancement expression of HSP expression and the downregulation of Apaf-1. Our studies suggest that combined PGA1 and Li may have potential clinical value for the treatment of stroke. [Abstract/Link to Full Text]

Kaur C, Dheen ST, Ling EA
From blood to brain: amoeboid microglial cell, a nascent macrophage and its functions in developing brain.
Acta Pharmacol Sin. 2007 Aug;28(8):1087-96.
Amoeboid microglial cells (AMC) in the developing brain are active macrophages. The macrophagic nature of these cells has been demonstrated by many methods, such as the localization of various hydrolytic enzymes and the presence of complement type 3 surface receptors in them. More importantly is the direct visualization of these cells engaged in the phagocytosis of degenerating cells at the ultrastructural level. Further evidence of them being active macrophages is the avid internalization of tracers administered by the intravenous or intraperitoneal routes in developing rats. The potential involvement of AMC in immune functions is supported by the induced expression of major histocompatibility complex class I and II antigens on them when challenged by lipopolysaccharide or interferon-gamma. Immunosuppressive drugs, such as glucocorticoids and immune function-enhancing drugs like melatonin, affect the expression of surface receptors and antigens and the release of cytokines by AMC. Recent studies in our laboratory have shown the expression of insulin-like growth factors, endothelins, 2',3'-cyclic nucleotide 3'-phosphodiesterase, and N-methyl-D-asparate receptors. This along with the release of chemokines, such as stromal derived factor-1a and monocyte chemoattractant protein-1, suggests multiple functional roles of AMC in early brain development. [Abstract/Link to Full Text]

Niu B, Lu WC, Yang SS, Cai YD, Li GZ
Support vector machine for SAR/QSAR of phenethyl-amines.
Acta Pharmacol Sin. 2007 Jul;28(7):1075-86.
AIM: To discriminate 32 phenethyl-amines between antagonists and agonists, and predict the activities of these compounds. METHODS: The support vector machine (SVM) is employed to investigate the structure-activity relationship (SAR)/quantitative structure-activity relationship (QSAR) of phenethyl-amines based on molecular descriptors. RESULTS: By using the leave-one-out cross-validation (LOOCV) test, 1 optimal SAR and 2 optimal QSAR models for agonists and antagonists were attained. The accuracy of prediction for the classification of phenethyl-amines by using the LOOCV test is 91.67%, and the accuracy of prediction for the classification of phenethyl-amines by using the independent test is 100%; the results are better than those of the Fisher, the artificial neural network (ANN), and the K-nearest neighbor models for this real world data. The RMSE (root mean square error) of antagonists' QSAR model is 0.5881, and the RMSE of agonists' QSAR model is 0.4779, which are better than those of the multiple linear regression, partial least squares, and ANN models for this real world data. CONCLUSION: The SVM can be used to investigate the SAR and QSAR of phenethylamines and could be a promising tool in the field of SAR/QSAR research. [Abstract/Link to Full Text]

Recent Articles in Biological & Pharmaceutical Bulletin

Sartoretto JL, Oliveira MA, Nigro D, Carvalho MH, Tostes RC, Fortes ZB
Constrictor responses to noradrenaline, hemodynamic profile, and superoxide levels measured by hydroethidine oxidation in diabetic rats.
Biol Pharm Bull. 2007 Oct;30(10):1938-42.
The aim of this study was to test the hypothesis that vascular dysfunction in neonatal streptozotocin (n-STZ)-induced diabetic rats could be associated with alterations in blood pressure, hemodynamic profile, and levels of superoxide anion. Diabetes was induced by STZ injection (160 mg/kg, i.p.) in neonate (2-d-old) Wistar rats. Using intravital microscopy the changes in mesenteric arteriolar diameters to vasoconstrictor agent noradrenaline (NA) and the levels of superoxide anion, measured by hydroethidine microfluorography, were determined in anaesthetized control and n-STZ rats. Blood pressure (BP) was determined in anaesthetized and unanaesthetized animals. Heart rate, shear rate, and blood flow velocity were also assessed. n-STZ rats showed, after 8 weeks of STZ injection, increased BP (unanaesthetized animals), hyperactivity to NA, and increased superoxide anion levels. However, heart rate, arteriolar shear rate, and blood flow velocity were unchanged in n-STZ. In conclusion, the results of the current study describe a significant increase in blood pressure, hyperactivity to NA-mediated vasoconstriction, and increased superoxide levels measured by hydroethidine oxidation. Taken together, our findings demonstrate that the compromised ability of mesenteric microvessels to respond properly in n-STZ diabetic rats is associated with several vascular alterations. [Abstract/Link to Full Text]

Nagai N, Fukuhata T, Ito Y, Tai H, Hataguchi Y, Nakagawa K
Preventive effect of water containing magnesium ion on paw edema in adjuvant-induced arthritis rat.
Biol Pharm Bull. 2007 Oct;30(10):1934-7.
We demonstrate the preventive effect of bittern water (BW), which enables the effective intake of magnesium ion (Mg(2+)), on paw edema in adjuvant-induced arthritis (AA) rat. BW (five kinds; BW-1, 2, 3, 4, 5) containing 10-200 mg/l Mg(2+) was used in this study. Arthritis was induced by the injection of 50 microl of a suspension of 10 mg/ml heat-killed butyricum (Mycobacterium butyricum) in Bayol F oil into the plantar region of the right hind foot and tail of rats. Paw edema of the right and left hind feet of AA rats were reduced by the administration of BW for 14 d after adjuvant injection in comparison with those of AA rats administered purified water. The preventive effect increased with the increasing Mg(2+) content of the BW. In addition, a combination of indomethacin (IM, 2 mg/kg) and BW-5 (200 mg/l Mg(2+)) prevented paw edema of the right and left hind feet of AA rats in comparison with IM alone. The fate of plasma IM after the oral administration of the combined IM (2 mg/kg/d) and BW-5 was similar to that after the administration of IM alone. In conclusion, the oral administration of Mg(2+) to AA rats potently prevents the development of inflammation, and the combination of IM and Mg(2+) may provide an effective therapy of arthritic edema. [Abstract/Link to Full Text]

Ishida K, Honda M, Shimizu T, Taguchi M, Hashimoto Y
Stereoselective metabolism of carvedilol by the beta-naphthoflavone-inducible enzyme in human intestinal epithelial Caco-2 cells.
Biol Pharm Bull. 2007 Oct;30(10):1930-3.
Treatment of Caco-2 cells with beta-naphthoflavone (beta-NF) and 1alpha,25-dihydroxyvitamin D(3) (VD(3)) induces UDP-glucuronosyltransferases (UGTs) and cytochrome P450 (CYP) 3A4, respectively. In the present study, we evaluated the metabolism of carvedilol in beta-NF- and VD(3)-treated Caco-2 cells. The metabolism of R-carvedilol was not significant in non-treated Caco-2 cells, whereas S-carvedilol was significantly metabolized in the cells. The metabolism of R- and S-carvedilol was significantly increased by the treatment of Caco-2 cells with 50 microM beta-NF for 3 d. In contrast, the treatment of Caco-2 cells with 250 nM VD(3) for 2 weeks did not induce a significant change in the metabolism of R- and S-carvedilol. The metabolism of carvedilol in beta-NF-treated Caco-2 cells was markedly inhibited by a substrate of UGTs, baicalein. In addition, the expression of UGT1A1, 1A6, and 1A9 mRNA was increased in beta-NF-treated Caco-2 cells as compared with non-treated cells. These findings indicated that carvedilol was metabolized stereoselectively by the beta-NF-inducible enzyme in Caco-2 cells. The UGT1A subfamily in intestinal epithelial cells may be partly responsible for first-pass (presystemic) metabolism of the drug. [Abstract/Link to Full Text]

Fujimoto S
Promising antitumor activity of a novel quinoline derivative, TAS-103, against fresh clinical specimens of eight types of tumors measured by flow cytometric DNA analysis.
Biol Pharm Bull. 2007 Oct;30(10):1923-9.
TAS-103, 6-[[2-(dimethylamino)ethyl]amino]-3-hydroxy-7H-indeno-[2,1-c]quinolin-7-one dihydrochloride, is a dual topoisomerases I and II inhibitor. Antitumor activities of TAS-103 against fresh surgical specimens resected from 525 patients (32 types of tumors) were examined by flow cytometric (FCM) analysis of DNA integrity of tumor cells, and compared with those of five other investigational new drugs and 31 clinically available anticancer agents. Concentrations of clinically available anticancer agents were set at one-tenth of the peak plasma concentration (PPC) of the clinically recommended doses. On the other hand, since PPCs of investigational new drugs in humans were frequently unknown, these were estimated by a method that determines the theoretically achievable concentration in body fluid (TAC method). Correlations between TAC and PPC were examined for 16 clinically available anticancer agents, and it was found that TAC at 7n (the modified Fibonacci's dose-escalation scheme) of 14 drugs corresponded well with each one-tenth of PPC. By defining a 30% or more reduction in the integrated diploid peak as effective and a 60% or more reduction as definitely effective, TAS-103 at 5 microg/ml (7n) showed significantly higher effective rates and definitely effective rates than those of all other investigational new drugs, as well as almost all clinically available anticancer agents, against various malignancies, including non-small cell lung cancer, brain tumor and renal cancer. These results strongly suggest that TAS-103 will be expected to show excellent antitumor activities against a wide range of human tumors. [Abstract/Link to Full Text]

Ling W, Rui LC, Hua JX
In situ intestinal absorption behaviors of tanshinone IIA from its inclusion complex with hydroxypropyl-beta-cyclodextrin.
Biol Pharm Bull. 2007 Oct;30(10):1918-22.
In this paper, the intestinal permeability of the inclusion complex of tanshinone IIA (TS IIA) with 2-hydroxypropyl-beta-cyclodextrin (HP-beta-CD) was investigated. The corresponding complexation of TS IIA-HP-beta-CD was obtained by coevaporation and characterized by differential scanning calorimetry and X-ray diffraction. The recirculation intestinal perfusion technique in rats was used to study the absorption behavior of free and complexed TS IIA. The change of concentration of TS IIA was separately calculated according to Michaelis-Menten and the Fick's equation to investigate its absorption rate-limiting step. Using the mathematical models above, it was concluded that the limit step to absorption of TS IIA was the dissolution process. Different concentrations of complexed TS IIA were administrated to three intestinal segments, with the intestinal permeability ranging from 3.16x10(-5) cm.s(-1) in the duodenum (50 microg.ml(-1)) to 4.11x10(-5) cm.s(-1) in the jejunum (100 microg.ml(-1)). With the increase of dosage of complex, TS IIA's absorption did not show saturated phenomenon, suggesting its transport mechanism in vivo might primary be passive transport. Besides, the permeability of TS IIA was not apparently influenced by the perfusion section studied, which indicated that there might not exist specific absorption site for TS IIA. [Abstract/Link to Full Text]

Kim HJ, Kim HJ
The glycosylation and pharmacokinetics of CTLA4Ig produced in rice cells.
Biol Pharm Bull. 2007 Oct;30(10):1913-7.
Cytotoxic T-lymphocyte antigen 4-immunoglobulin (CTLA4Ig) has immunosuppressive activity and the ability to induce immune tolerance. There has been no report of its glycosylation ratio or of the role of its glycans. We investigated the terminal sialylation of rice cell-derived recombinant human CTLA4Ig (rrhCTLA4Ig) using lectins. The glycosylation ratios of rrhCTLA4Ig and Chinese hamster ovary (CHO) cell-derived recombinant human CTLA4Ig (crhCTLA4Ig) were evaluated by chemical deglycosylation. After intravenous (i.v.) or subcutaneous (s.c.) administration to rats, the pharmacokinetics of rrhCTLA4Ig and crhCTLA4Ig as well as of their deglycosylated forms were evaluated. rrhCTLA4Ig does not have terminal sialic acids and its glycosylation ratio was slightly lower than that of crhCTLA4Ig. Its terminal elimination half-life (T(1/2)) was shorter than that of crhCTLA4Ig following i.v. administration. However, the half-life was significantly prolonged and was similar with that of crhCTLA4Ig following s.c. administration. Moreover, the deglycosylated forms of both preparations were cleared from the circulation faster than the native forms. These results suggest that the presence of glycans on rrhCTLA4Ig and crhCTA4Ig are important for their in vivo stability. In addition, the glycan structure of rrhCTLA4Ig is more effective in maintaining in vivo stability after s.c. administration than after i.v. administration although the glycans on rrhCTLA4Ig lack terminal sialic acids, suggesting that its glycans have the potential for in vivo stability. [Abstract/Link to Full Text]

Takayama J, Takaoka M, Sugino Y, Yamamoto Y, Ohkita M, Matsumura Y
Sex difference in ischemic acute renal failure in rats: approach by proteomic analysis.
Biol Pharm Bull. 2007 Oct;30(10):1905-12.
It is known that female rats are resistant to ischemic acute renal failure (ARF), compared with male rats. To elucidate sex differences in ischemic ARF, we searched global protein expression in post-ischemic kidneys using proteomic techniques. Ischemic ARF was induced by 45-min ischemia followed by reperfusion. By proteomic analysis, many male- or female-dominant proteins were detected in sham-operated rat kidneys, and significantly increased or decreased proteins were found in post-ischemic kidneys 2 h after reperfusion, at which there were no significant deterioration in renal function of both sexes. We detected 86 proteins showing more than 1.5-fold significant alterations (p<0.01) in both sexes by ischemia/reperfusion (I/R) treatment. Among the altered proteins, we identified a significantly up-regulated protein in male rat kidneys, meprin alpha, a subunit of meprin which had been reported to play a role in the pathophysiology of I/R-induced ARF. In addition, it is known that a potent meprin alpha inhibitor, actinonin, can protect against I/R-induced renal injury when administered to male rats. We therefore compared the effect of actinonin on I/R-induced renal dysfunction between male and female rats. Renal function of both males and females showed significant deterioration when measured at 24 h after the reperfusion, although the degree of renal dysfunction was much less in females than in males. Pre-ischemic treatment with actinonin (30 mg/kg, i.v.) prevented the I/R-induced renal dysfunction in males but not in females. Our results provide information on differences in protein expression at an early phase after the reperfusion between male and female rats. Moreover, the present study suggests that up-regulation of meprin alpha in the post-ischemic kidney is at least partly involved in aggravation of I/R-induced renal injury in male rats. The possibility that meprin alpha is a key component of the sex difference in ischemic ARF, warrants further attention. [Abstract/Link to Full Text]

Lee CH, Park SW, Kim YS, Kang SS, Kim JA, Lee SH, Lee SM
Protective mechanism of glycyrrhizin on acute liver injury induced by carbon tetrachloride in mice.
Biol Pharm Bull. 2007 Oct;30(10):1898-904.
Glycyrrhizin is the major active component extracted from licorice (Glycyrrhiza glabra) roots, one of the most widely used herbal preparations for the treatment of liver disorders. This study evaluated the potential beneficial effect of glycyrrhizin in a mouse model of carbon tetrachloride (CCl(4))-induced liver injury. The mice were treated intraperitoneally with CCl(4) (0.5 ml/kg). They received glycyrrhizin (50, 100, 200, 400 mg/kg) 24 h and 0.5 h before and 4 h after administering CCl(4). The serum activities of aminotransferase and the hepatic level of malondialdehyde were significantly higher 24 h after the CCl(4) treatment, while the concentration of reduced glutathione was lower. These changes were attenuated by glycyrrhizin. CCl(4) increased the level of circulating tumor necrosis factor-alpha markedly, which was reduced by glycyrrhizin. The levels of hepatic inducible nitric oxide synthase, cyclooxygenase-2, and heme oxygenase-1 protein expression were markedly higher after the CCl(4) treatment. Glycyrrhizin diminished these alterations for inducible nitric oxide and cyclooxygenase-2 but the protein expression of heme oxygenase-1 was further elevated by the treatment of glycyrrhizin. CCl(4) increased the level of tumor necrosis factor-alpha, inducible nitric oxide synthase, cyclooxygenase-2, and heme oxygenase-1 mRNA expressions. The mRNA expression of heme oxygenase-1 was augmented by the glycyrrhizin treatment, while glycyrrhizin attenuated the increase in tumor necrosis factor-alpha, inducible nitric oxide synthase, and cyclooxygenase-2 mRNA expressions. These results suggest that glycyrrhizin alleviates CCl(4)-induced liver injury, and this protection is likely due to the induction of heme oxygenase-1 and the downregulation of proinflammatory mediators. [Abstract/Link to Full Text]

Uchida M, Suzuki M, Shimizu K
Effects of urocortin, corticotropin-releasing factor (CRF) receptor agonist, and astressin, CRF receptor antagonist, on the sleep-wake pattern: analysis by radiotelemetry in conscious rats.
Biol Pharm Bull. 2007 Oct;30(10):1895-7.
Stress has been known to release corticotropin-releasing factor (CRF) and have an affect on sleep-wake patterns. However, there is no direct evidence of CRF receptor agonist and antagonist on sleep-wake patterns. Therefore, this study aimed to clarify this point by using radiotelemetry system in conscious rats. Wake, non-rapid eye-moving (NREM) sleep and rapid eye-moving (REM) sleep were analyzed by computer software, simultaneously measuring electroencephalogram and electromyogram. In the light period, urocortin (CRF receptor agonist: i.v.) significantly increased wake duration, and decreased NREM sleep duration. REM sleep was not affected. Astressin (CRF receptor antagonist: i.p.) significantly attenuated the changes induced by urocortin, although astressin itself did not affect the sleep-wake pattern in the light period at this dosage. These findings show that urocortin changes the sleep-wake pattern in the light period. Moreover, urocortin was found to change the sleep-wake pattern by acting on CRF receptor, as astressin significantly attenuated the urocortin-induced changes on sleep-wake patterns. [Abstract/Link to Full Text]

Takahashi K, Hossain M, Ahmed M, Bhuiyan MA, Ohnuki T, Nagatomo T
Asp125 and Thr130 in transmembrane domain 3 are major sites of alpha1b-adrenergic receptor antagonist binding.
Biol Pharm Bull. 2007 Oct;30(10):1891-4.
Site-directed mutagenesis was used to investigate the molecular interactions involved in prazosin binding to the human alpha(1b)-adrenergic receptor (alpha(1b)-AR) receptor. Based on molecular modeling studies, Thr130 and Asp125 in transmembrane region III of the alpha(1b)-AR receptor were found to interact with prazosin. Thr130 and Asp125 were mutated to alanine (Ala) and expressed in HEK293 cells. The radioligand [(3)H]prazosin did not show any binding to Asp125Ala mutant of alpha(1b)-AR. Therefore, it was not possible to find any prazosin affinity to the mutant using the radioligand [(3)H]prazosin. The mutation also abolished phenylephrine-stimulated inositol phosphate (IP) formation of [(3)H]myo-inositol. On the other hand, the Thr130Ala mutant showed reduced binding affinity for [(3)H]prazosin (dissociation constant, K(d) 674.27 pM versus 90.27 pM for the wild-type receptor) and had reduced affinity for both tamsulosin and prazosin (11-fold and 9-fold, respectively). However, the Thr130Ala mutant receptor retained the ability to stimulate the formation of [(3)H]myo-inositol. The results provide direct evidence that Asp125 and Thr130 are responsible for the interactions between alpha(1b)-AR receptor and radioligand [(3)H]prazosin as well as tamsulosin. [Abstract/Link to Full Text]

Yang X, Zhao Y, Zhou Y, Lv Y, Mao J, Zhao P
Component and antioxidant properties of polysaccharide fractions isolated from Angelica sinensis (OLIV.) DIELS.
Biol Pharm Bull. 2007 Oct;30(10):1884-90.
An analytical method of high performance capillary electrophoresis (HPCE) was developed to simultaneously separate and identify the component monosaccharides of Angelica sinensis polysaccharide fractions (APFs), named APF1, APF2 and APF3. The predominant sugars in APFs were identified as arabinose, glucose, rhamnose, galactose and galacturonic acid as well as trace amount of mannose and glucuronic acid, and the fractionation altered significantly the distribution of component monosaccharides in APFs. APF3 was the most active fraction to effectively inhibit H(2)O(2)-caused decrease of cell viability, lactate dehydrogenase (LDH) leakage and malondialdehyde (MDA) formation, and also reduced H(2)O(2)-caused decline of superoxide dismutase (SOD) activity and glutathione (GSH) depletion (p<0.05), followed by APF2 and APF1 in decreasing order. Furthermore, it was found that APFs (100 microg/ml) could protect macrophages by inhibiting the release of excess NO and reactive oxygen species (ROS) induced by high concentrations of H(2)O(2) (0.8-1.6 mM). [Abstract/Link to Full Text]

Nishimura R, Tabata K, Arakawa M, Ito Y, Kimura Y, Akihisa T, Nagai H, Sakuma A, Kohno H, Suzuki T
Isobavachalcone, a chalcone constituent of Angelica keiskei, induces apoptosis in neuroblastoma.
Biol Pharm Bull. 2007 Oct;30(10):1878-83.
Six chalcones from Angelica keiskei KOIDZUMI (Ashitaba in Japanese) and two chalcones from Humulus lupulus L. (hop) were examined for their cytotoxicity in two human neuroblastoma cell lines (IMR-32 and NB-39) and normal cells (primary culture of rat cerebellar granule cells) by [3-(4,5)-dimethyl-2-thiazolyl]-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. All chalcones exhibited cytotoxicity against neuroblastoma cells, and two of them (isobavachalcone and xanthoangelol H) had no effect on normal cells even at high concentration (10(-4) M) exposure. Typical morphologic features of apoptosis, including cell shrinkage, chromatin condensation, nuclear fragmentation and formation of apoptotic bodies, were observed in isobavachalcone-treated cells by Hoechst 33342 staining. Western blot analysis showed that isobavachalcone significantly reduced pro-caspase-3 and pro-caspase-9, and subsequently increased the level of cleaved caspase-3 and cleaved caspase-9 in both neuroblastoma cell lines. Moreover, Bax was markedly induced by isobavachalcone application. These results suggest that isobavachalcone induces apoptotic cell death in neuroblastoma via the mitochondrial pathway and has no cytotoxicity against normal cells. Therefore, isobavachalcone may be applicable as an efficacious and safe drug for the treatment of neuroblastoma. [Abstract/Link to Full Text]

Iwatsuki Y, Sakata C, Kawasaki T, Okada M
Experimental model of lower limb ischemia in rats and the effect of YM466, an oral direct factor Xa inhibitor.
Biol Pharm Bull. 2007 Oct;30(10):1874-7.
A simple, quantitative, and reproducible model of lower limb ischemia was developed. Vascular injury was induced by ferric chloride (FeCl(3)) solution to the rat iliac artery, after which blood flow in all of the lower limbs were continuously monitored using a scanning laser Doppler blood flowmeter. After FeCl(3) injury, a distinct decrease in blood flow in the ischemic lower limb was observed and blood flow did not recover during the 30 min after vascular injury. YM466, an oral direct factor Xa inhibitor, dose-dependently inhibited the reduction of peripheral blood flow. The area under the blood flow-time curve during 30 min after vascular injury improved dose-dependently, with significance at doses of 3 and 10 mg/kg. These results suggest that factor Xa inhibitors are effective in patients with peripheral arterial disease, and that this vascular injury model is a useful tool for the screening and evaluation of the efficacy of new antithrombotic agents. [Abstract/Link to Full Text]

Okawa Y, Miyauchi M, Takahashi S, Kobayashi H
Comparison of pathogenicity of various Candida albicans and C. stellatoidea strains.
Biol Pharm Bull. 2007 Oct;30(10):1870-3.
In order to clarify the pathogenicity and the pathogenic factors of various Candida species strains, three strains, NIH A-207 and J-1012 (serotype A), and NIH B-792 (serotype B) of Candida albicans and two strains, ATCC 20408 (karyotype II) and ATCC 36232 (karyotype I) of C. stellatoidea, a synonym for C. albicans, were tested for their lethality to mice, adherence to Hela cells, hydrophobicity, and cell growth under acidic conditions, pH 2.0-5.9. The pathogenicity for mice of all the strains was observed in the order NIH B-792, ATCC 36232, J-1012, NIH A-207, and ATCC 20408. The pathogenicity for mice by all the strains used was well correlated with adherence to the Hela cells, the hydrophobicity, and the cell growth under the acidic condition, pH 2.0. These results emphasize that these specific properties of the C. albicans and C. stellatoidea strains play an important role in the pathogenesis of candidosis. [Abstract/Link to Full Text]

Sung WS, Lee DG
In vitro antimicrobial activity and the mode of action of indole-3-carbinol against human pathogenic microorganisms.
Biol Pharm Bull. 2007 Oct;30(10):1865-9.
Indole-3-carbinol (I3C) is a naturally occurring constituent of cruciferous vegetables. The aim of this study was to assess the in vitro antimicrobial activity of I3C and its mode of action. By using an NCCLS broth microdilution assay, the activity of I3C was evaluated against human pathogenic microorganisms including clinically isolated antibiotic-resistant bacterial strains. The results indicated that I3C exhibited broad spectrum antimicrobial activities. To elucidate the physiological changes of the fungal cells induced by I3C, we performed a flow cytometric analysis for a cell cycle. The results showed that I3C arrested the cell cycle at the G(2)/M phase in Candida albicans. To understand the antifungal mode of action of I3C, the change in the membrane dynamics was monitored by using fluorescence changing experiments against C. albicans. The results suggest that I3C may exert antifungal activity by disrupting the structure of the cell membrane. The present study indicates that I3C has considerable antimicrobial activity, deserving further investigation for clinical applications. [Abstract/Link to Full Text]

Ohbayashi N, Ikeda O, Taira N, Yamamoto Y, Muromoto R, Sekine Y, Sugiyama K, Honjoh T, Matsuda T
LIF- and IL-6-induced acetylation of STAT3 at Lys-685 through PI3K/Akt activation.
Biol Pharm Bull. 2007 Oct;30(10):1860-4.
Signal transducer and activator of transcription 3 (STAT3), which mediates biological actions in many physiological processes, is activated by cytokines and growth factors via specific tyrosine or serine phosphorylation, dimerization and nuclear translocation. A recent study has demonstrated, by using antibody to acetylated lysine, and a STAT3 mutant with Lys-685-to-Arg substitution, that STAT3 is acetylated at Lys-685 by histone acetyltransferase p300, and that acetylation at Lys-685 is critical for STAT3 activation. In the present study, we created an acetyl-specific antibody against STAT3 acetylated at Lys-685, and found that leukemia inhibitory factor (LIF) or interleukin (IL)-6 induced acetylation of STAT3 at Lys-685 in 293T and Hep3B cells. Moreover, acetylation of STAT3 at Lys-685 was suppressed by PI3K inhibitor LY294002, or a dominant negative Akt. Taken together, our findings demonstrate that endogenous STAT3 is acetylated at Lys-685 by LIF or IL-6 through PI3K/Akt activation. [Abstract/Link to Full Text]

Nagamine K, Furue M, Fukui A, Matsuda A, Hori T, Asashima M
Blood cell and vessel formation following transplantation of activin-treated explants in Xenopus.
Biol Pharm Bull. 2007 Oct;30(10):1856-9.
Treatment of Xenopus blastula with activin converts undifferentiated presumptive ectoderm (animal cap) into mesoderm and endoderm in a dose-dependent manner. At low concentrations, activin induces ventral mesoderm such as blood cells. Here we show that activin-treated aggregates of animal cap cells prepared from undifferentiated presumptive ectoderm and transplanted into Xenopus embryos differentiated to form red blood cells and vascular endothelial cells. We compared gene expression profiles of the activin-treated with untreated aggregates of animal cap cells using microarray analysis. This revealed 838 clones including vascular-related genes that were expressed at levels at least 2-fold greater in the activin-treated aggregates than in the untreated controls. Of these, 356 were known Xenopus genes, 296 had homologues, and 186 were unknown genes. These findings identified novel vascular-related genes and provided insights into how the blood vessel system establishes in normal development. [Abstract/Link to Full Text]

Huaping L, Jie L, Zhifeng W, Yingchang Z, Yuhuan L
Cloning and functional expression of ubiquitin-like protein specific proteases genes from Candida albicans.
Biol Pharm Bull. 2007 Oct;30(10):1851-5.
The small ubiquitin-like modifier (SUMO) modification occurred at bud necks and sites of septum formation in hyphae of the opportunistic fungal pathogen Candida albicans. Three genes encoding putative SUMO deconjugation enzymes (Ulp, ubiquitin-like protein specific proteases) of C. albicans were obtained through sequence database searching with Ulp domain of Saccharomyces cerevisiae Ulp1 (ScUlp1). These genes were designated as CaULP1, CaULP2 and CaULP3. The open reading frames of three putative ULPs were cloned and expressed in Pichia pastoris, resulting recombinant proteins. Functional analysis of recombinant CaUlp1, CaUlp2 and CaUlp3 confirms that these proteins exhibit SUMO-processing activity. CaULP1, CaULP2 and CaULP3 only expressed active form enzyme in P. pastoris but not in Escherichia coli. The molecular weights of CaUlp1, CaUlp2 and CaUlp3 proteins expressed in P. pastoris were larger than theoretical molecular weights. This observation was in good agreement with result of Western blot analysis of CaUlp1 and CaUlp3 proteins in C. albicans. It was assumed that CaUlp1, CaUlp2 and CaUlp3 proteins may need post-translational modifications to exhibit SUMO-processing activity. To our knowledge, this is the first report on cloning and expression of Ulp genes from C. albicans. Furthermore RT-PCR and Western blot analysis show that CaULP2 has no detectable expression both in yeast and in hyphal forms of C. albicans. [Abstract/Link to Full Text]

Iijima A, Hachisu R, Kobayashi H, Hashimoto K, Asano D, Kikuchi H
Establishment of evaluation method for siRNA delivery using stable cell line carrying the luciferase reporter gene.
Biol Pharm Bull. 2007 Oct;30(10):1844-50.
We determined the influence of siRNA (short interfering RNA) for expression of plasmid DNA (pDNA), when mismatched siRNA and pDNA encoding beta-galactosidase (beta-gal) were transfected into HeLa cells by the cotransfection method in which they were simultaneously added to the cells. Cationic liposomes (Lipofectamine2000) were used as a gene transfection reagent. The knockdown effect on beta-gal was observed even when mismatched siRNA was used, and the effect depended on the amount of added mismatched siRNA. But, there was not a distinct difference of introduction of pDNA into cells between using mismatched siRNA and without using it. We considered that the cotransfection method should be avoided when we confirm RNAi efficiency. The reliable evaluation method for siRNA delivery in vitro was thus established by using NFAT reporter HeLa stable cell line or CHO (pMAM-luc) cell line that had DNA encoding luciferase. The following experimental conditions for each cell line were optimized: cell numbers seeded, total incubation times, concentrations of added inducers, and incubation times after addition of inducers. Transfection performance was compared for six commercially available reagents by this method. No commercially available transfection reagent, however, could reduce luciferase activity by less than one tenth without causing cellular cytotoxicity. Development of novel reagents providing higher transfection effects without cytotoxicity is needed. [Abstract/Link to Full Text]

Ito T, Sugita Y, Takao K, Ikeguchi Y, Shirahata A
Amine modification of digested peptide at C-terminal end during protein digestion by protease.
Biol Pharm Bull. 2007 Oct;30(10):1838-43.
We recently reported that C-terminal polyamine modification occurs when proteins are digested with trypsin in the presence of polyamine [Biochem. Biophys. Res. Commun., 356, 159-162 (2007)]. In the present study, the characteristics of this C-terminal modification in the presence of protease and amine were investigated. When hemoglobin (HB) was digested with trypsin in the presence of N-(2-pyridyl)-1,4-diaminobutane (Py4), formation of the modified peptide was dependent on time and on HB or Py4 concentration. When synthetic peptide was treated with trypsin in the presence of Py4, ca. 0.1% of the peptide was modified with Py4. When HB or cytochrome C was treated with a range of serine proteases in the presence of various amines (Py4, N-(2-pyridyl)-1,3-diaminopropane, tranexamic acid, isonicotinic acid hydrazide and ampicillin), the modified peptide was detected in all cases tested, thus suggesting that amine modification widely accompanies digestion by proteases. [Abstract/Link to Full Text]

Higai K, Sano R, Satake M, Azuma Y, Matsumoto K
Glycated human serum albumin induces interleukin 8 mRNA expression through reactive oxygen species and NADPH oxidase-dependent pathway in monocyte-derived U937 cells.
Biol Pharm Bull. 2007 Oct;30(10):1833-7.
Glycated human serum albumin (Glc-HSA) has previously been reported (Higai K., et al., 2006) to induce E-selectin expression on human umbilical vein endothelial cells through activation of NADPH oxidase; however, Glc-HSA signaling in monocytes remains obscure. To clarify the influence on human monocyte-derived U937 cells, U937 cells were stimulated with Glc-HSA and glycoaldehyde-dimer-modified HSA (GA-HSA) for 2 h in the absence and presence of the protein kinase C (PKC) inhibitor calphostin and the reactive oxygen species (ROS) scavenger N-acetyl-L-cysteine (NAC) and NADPH oxidase inhibitor apocynin; interleukin-8 (IL-8) mRNA expression was determined by RT-PCR. As a result, IL-8 mRNA expression in U-937 cells was time- and dose-dependently enhanced by stimulation with Glc-HSA and GA-HSA. Furthermore, promoter activity of the IL-8 reporter gene was enhanced approximately 2-fold by stimulation with Glc-HSA and GA-HSA. Nuclear factor kappaB (NFkappaB) and activator protein-1 (AP-1) reporter genes were also enhanced although CCAAT/enhancer binding protein (C/EBP) was not affected. IL-8 mRNA expression was suppressed by NAC and apocynin but not calphostin in cells stimulated with Glc-HSA; however, its expression in cells stimulated with GA-HSA was suppressed by calphostin but not NAC. These results indicated that IL-8 mRNA expression was upregulated by NFkappaB and AP-1 in U937 cells stimulated with Glc-HSA and GA-HSA, but the signaling pathways were different. [Abstract/Link to Full Text]

Yoshida N, Aizu-Yokota E, Sonoda Y, Moriwaki Y, Kishi K, Kasahara T
Production and regulation of eotaxin-2/CCL24 in a differentiated human leukemic cell line, HT93.
Biol Pharm Bull. 2007 Oct;30(10):1826-32.
When a human leukemic cell line, HT93 was incubated with all-trans retinoic acid (ATRA), IL-5, or both, this cell line was differentiated into eosinophic lineage, in that an eosinophilic specific granule proteins, major basic protein (MBP) and eosinophil peroxidase (EPO) appeared. Both CD11b and CC chemokine receptor, CCR3 expression were upregulated, while CD71 expression was downregulated by ATRA or ATRA+IL-5. Concomitantly, marked production of eotaxin-2/CCL24 was observed, but no production of eotaxin-1/CCL11 and eotaxin-3/CCL26 was detected. Since only 20 to 30% cells incubated with ATRA became positive for CCR3, CCR3(+) population was enriched by a magnetic activated cell sorter (MACS). Enriched CCR3(+) population produced higher eotaxin-2/CCL24 than the CCR3(-) population, indicating that differentiated eosinophils are capable of producing eotaxin-2/CCL24. During the ATRA-induced differentiation, expression of a transcriptional factor, GATA-1 was significantly increased. Introduction of siRNA against GATA-1 markedly reduced the ATRA-induced differentiation markers including CD11b and CCR3, as well as reduced eotaxin-2/CCL24 production. Finally, ATRA-induced differentiation and eotaxin-2/CCL24 production were greatly enhanced in the GATA-1-overexpressed clones. These results indicate that the ability to produce eotaxin-2/CCL24 is acquired during the differentiation into eosinophilic lineage which is dependent on GATA-1 expression. [Abstract/Link to Full Text]

Itoh N
The Fgf families in humans, mice, and zebrafish: their evolutional processes and roles in development, metabolism, and disease.
Biol Pharm Bull. 2007 Oct;30(10):1819-25.
Fibroblast growth factors (Fgfs) were originally isolated as growth factors for fibroblasts. However, Fgfs are now recognized as polypeptide growth factors of ca. 150-250 amino acid residues with diverse biological activities and expression profiles. The Fgf signaling system has been identified in multicelluar but not in unicellular organisms. In contrast to the only two Fgf genes and one Fgf receptor (Fgfr) gene in Caenorhabditis elegans, both the human and mouse Fgf and Fgfr gene families comprise twenty-two and four members, respectively. Their evolutional processes indicate that the Fgf and Fgfr gene families greatly co-expanded during the evolution of early vertebrates. The expansion of the Fgf and Fgfr gene families has enabled this signaling system to acquire diversity of function and a nearly ubiquitous involvement in many developmental and physiological processes. The zebrafish fgf gene family comprises twenty-seven members with several paralogs generated by an additional genome duplication. The mouse and zebrafish are useful models for studying gene functions. Fgf knockout mice have been generated. Several Fgf knockout mice die in the embryonic or early postnatal stages, indicating crucial roles for these genes in various developmental processes. However, other Fgf knockout mice survive with subtle phenotypic alterations. Their functions might be redundant. Studies using zebrafish embryos with mutated or knockdown fgfs also indicate that fgfs play crucial roles in development in that species. Although most Fgfs act in development in a paracrine and/or autocrine manner, some have potential roles in metabolism in an endocrine manner. In humans, Fgf signaling disorders result in hereditary diseases and cancers. [Abstract/Link to Full Text]

Satoh K, Nonaka R, Ogata A, Nakae D, Uehara S
Effects of oseltamivir phosphate (Tamiflu) and its metabolite (GS4071) on monoamine neurotransmission in the rat brain.
Biol Pharm Bull. 2007 Sep;30(9):1816-8.
As abnormal behaviors such as jumping and falling from balcony were reported in patients aged 10 to 19 years who administrated oseltamivir phosphate (Tamiflu) for treatment influenza infection, the Ministry of Health, Labor and Welfare in Japan notified that, as a rule, Tamiflu should not be prescribed to patients aged 10 to 19 years. To examine the relationship between Tamiflu and abnormal behaviors, we investigated the effects of Tamiflu and its carboxylic acid metabolite, GS4071, on the central nervous system, that is, on 3 neurotransmitters (dopamine, serotonin, and norepinephrine) in presynapses (inhibition of re-uptake, promotion of release) and postsynapses (guanosine 5'-triphosphate (GTP) gammaS binding), using rat brain synaptosomes. Neither Tamiflu nor GS4071 influenced the re-uptake/release of the 3 monoamines or GTP binding in postsynapses. [Abstract/Link to Full Text]

Mendes A, Mores AU, Carvalho AP, Rosa RT, Samaranayake LP, Rosa EA
Candida albicans biofilms produce more secreted aspartyl protease than the planktonic cells.
Biol Pharm Bull. 2007 Sep;30(9):1813-5.
By using a simple, low-cost system of polystyrene centrifuge tubes we compared the secreted aspartyl proteases (Saps) secretion during the biphasic growth modes of Candida albicans using twenty-one clinical isolates. Our results indicate that biofilms of C. albicans consistently secrete more Saps than their planktonic counterparts. [Abstract/Link to Full Text]

Moon Y, Kim HK, Suh H, Chung DH
Toxic alterations in chick embryonic liver and spleen by acute exposure to Fusarium-producing mycotoxin deoxynivalenol.
Biol Pharm Bull. 2007 Sep;30(9):1808-12.
Food mycotoxin deoxynivalenol (vomitoxin, DON) produced by Fusarium graminearum and F. culmorum can induce rapid diminution of lymphoid tissues and lymphopenia in the growing chickens and mammals. We first investigated the direct acute effects of DON on the chick immune-related embryo tissues such as embryonic liver and spleen. Direct DON administration into the embryonic eggs caused toxin accumulation in liver in a time-dependent manner. Electron microscopic observation showed a notable accumulation of fat droplet in the liver tissue and the re-exposed hatched chicken showed more distinguishing enlarged fat globules, so-called fatty cysts like human steatosis. Regarding effects of deoxynivalenol on the chick embryonic spleen, fatty change was also observed in splenocytes. Functionally, mitogen-stimulated cellular and humoral lympho-proliferations were suppressed in the DON-treated embryo. Conclusively, acute direct exposure to deoxynivalenol in the chick embryo caused toxic histological alterations in the liver and spleen and suppressed in vitro lymphoblastogenesis. [Abstract/Link to Full Text]

Shahverdi AR, Fakhimi A, Zarrini G, Dehghan G, Iranshahi M
Galbanic acid from Ferula szowitsiana enhanced the antibacterial activity of penicillin G and cephalexin against Staphylococcus aureus.
Biol Pharm Bull. 2007 Sep;30(9):1805-7.
In this study the enhancement effect of Ferula szowitsiana roots' acetone extract on the antibacterial activity of penicillin G and cephalexin was evaluated against Staphylococcus aureus. Disk diffusion and broth dilution methods were used to determine the antibacterial activity of these antibiotics in the absence and presence of plant extract and its various fractions separated by TLC plate. The active component of plant extract involved in enhancement of penicillin G's and cephalexin's activities had Rf=0.336 on a TLC plate. The spectral data ((1)H-, (13)C-NMR) of this compound revealed that this compound was 7-[6-(beta-carboxyethyl)-5-isopropylidene-1,2-dimethylcyclo-hexylmethoxy]coumarin (galbanic acid), previously isolated from Ferula assa-foetida. In the presence of sub-inhibitory concentration of galbanic acid (100 microg/ml) the MIC of penicillin G for S. aureus decreased from 64 to 1 (a sixteen four-fold decrease) and for cephalexin from 128 to 1 microg/ml (a one hundred twenty eight-fold decrease). The highest fold decrease in MIC was observed for cephalexin in combination of galbanic acid against test strain. These results signify that the low concentration of galbanic acid (100 microg/ml) potentiates the antimicrobial action of penicillin G and cephalexin suggesting a possible utilization of these compounds in combination therapy against S. aureus. [Abstract/Link to Full Text]

Asakura Y, Kurosaki F
Cloning and expression of Dcga gene encoding alpha subunit of GTP-binding protein in carrot seedlings.
Biol Pharm Bull. 2007 Sep;30(9):1800-4.
A homology-based cloning strategy yielded a cDNA clone designated Dcga, presumably encoding alpha subunit of GTP-binding protein, from carrot (Daucus carota) seedlings. Molecular phylogenetic tree analysis of G protein alpha subunits from various biological sources suggested that, unlike in animal cells, classification of Galpha proteins into specific subfamilies could not be applicable to the proteins from higher plants. The restriction digests prepared from genomic DNA of carrot showed one or two hybridized signals in Southern blot analyses, and the expression level of Dcga was appreciably decreased upon the exposure of carrot to high temperature or the prolonged treatment with salt. These results suggest that Dcga occurs as single or double copy genes in carrot genome, and its transcript might play specific roles in heat- and salt-induced responses of the plant. [Abstract/Link to Full Text]

Kang DG, Moon MK, Lee AS, Kwon TO, Kim JS, Lee HS
Cornuside suppresses cytokine-induced proinflammatory and adhesion molecules in the human umbilical vein endothelial cells.
Biol Pharm Bull. 2007 Sep;30(9):1796-9.
Cornuside is a bisiridoid glucoside compound isolated from the fruit of Cornus officinalis SIEB. et ZUCC. The present study was designed to examine the effects of cornuside on expression levels of cytokine-induced proinflammatory and adhesion molecules in the human umbilical vein endothelial cells (HUVECs). Cornuside treatment attenuated tumor necrosis factor-alpha (TNF-alpha)-induced nuclear factor-kappa B (NF-kappaB) p65 translocation in HUVECs. In addition, cornuside suppressed the expression levels of endothelial cell adhesion molecules including intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) induced by TNF-alpha. TNF-alpha-induced monocyte chemoattractant protein 1 (MCP-1) expression was also attenuated by treatment of cornuside. These inhibitory effects of cornuside on proinflammatory and adhesion molecules were not due to decreased HUVEC viability as assessed by MTT test. Taken together, the present study suggests that cornuside suppresses expression levels of cytokine-induced proinflammatory and adhesion molecules in the human endothelial cells. [Abstract/Link to Full Text]

Kawauchi K, Tobiume K, Kaneko S, Kaneshiro K, Okamoto S, Ueda E, Kamata H, Moriyama Y, Hirata H
Suppression of AP-1 activity by cycloprodigiosin hydrochloride.
Biol Pharm Bull. 2007 Sep;30(9):1792-5.
Cycloprodigiosin hydrochloride (cPrG.HCl), a compound isolated from a marine bacterium, acts as an immunosuppressant and an anti-cancer drug. We have previously reported that cPrG.HCl suppressed the transcriptional activation of nuclear factor (NF)-kappaB. Here we studied the effect of cPrG.HCl on activation of another transcription factor, activator protein 1 (AP-1). cPrG.HCl potently suppressed AP-1 activity induced by tumor necrosis factor (TNF) alpha and phorbol myristate acetate (PMA). cPrG.HCl did not inhibit any of the mitogen-activated protein kinase (MAPK) families, whereas it did suppress transcriptional activation of AP-1 induced by constitutively activated mutants of MEKK1 or Ras. cPrG.HCl inhibited neither TNFalpha- or PMA-induced DNA-binding of AP-1 nor co-activator p300-induced activation of AP-1. Taken together, cPrG.HCl suppresses AP-1-dependent gene expression downstream of MAPK group through the inhibition of the transcription activation step of the AP-1 promoter complex. [Abstract/Link to Full Text]

Recent Articles in Chemical & Pharmaceutical Bulletin (Tokyo)

Li H, Tanaka T, Zhang YJ, Yang CR, Kouno I
Rubusuaviins A-F, monomeric and oligomeric ellagitannins from Chinese sweet tea and their alpha-amylase inhibitory activity.
Chem Pharm Bull (Tokyo). 2007 Sep;55(9):1325-31.
Six new ellagitannins herein, rubusuaviins A-F, were isolated from the aqueous acetone extract of Chinese sweet tea (Tien-cha, dried leaves of Rubus suavissimus S. LEE) together with seven known tannins. Rubusuaviin A was characterized as 1-O-galloyl-2,3-O-(S)-HHDP-4,6-O-(S)-sanguisorboyl-beta-D-glucopyranose. Rubusuaviins B, C, and E are dimeric, trimeric, and tetrameric ellagitannins, respectively, in which the sanguisorboyl groups were connected ellagitannin units. Rubusuaviins D and F were desgalloyl derivatives of rubusuaviins C and E, respectively. The inhibition of alpha-amylase activity by rubusuaviins and related ellagitannins was compared. Ellagitannins with beta-galloyl groups at the glucose C-1 positions showed stronger inhibition compared with the alpha-galloyl and desgalloyl compounds. The molecular weight of these compounds was not important for the inhibition of alpha-amylase activity. [Abstract/Link to Full Text]

Mimaki Y, Doi S, Kuroda M, Yokosuka A
Triterpene glycosides from the stems of Akebia quinata.
Chem Pharm Bull (Tokyo). 2007 Sep;55(9):1319-24.
The stems of Akebia quinata have been analyzed for their triterpene glycoside constituents, resulting in the isolation of six new triterpene glycosides, along with 19 known ones. On the basis of extensive spectroscopic analysis, including 2D NMR data, and chemical evidence, the structures of the new compounds were deter-mined to be 3beta-[(O-beta-D-glucuronopyranosyl-(1-->3)-alpha-L-arabinopyranosyl)oxy]olean-12-en-28-oic acid O-alpha-L-rhamnopyranosyl-(1-->4)-O-beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranosyl ester, 3beta-[(O-beta-D-glucuronopyranosyl-(1-->3)-O-[alpha-L-rhamnopyranosyl-(1-->2)]-alpha-L-arabinopyranosyl)oxy]olean-12-en-28-oic acid O-alpha-L-rhamnopyranosyl-(1-->4)-O-beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranosyl ester, 3beta-[(O-beta-D-glucuronopyranosyl-(1-->3)-alpha-L-arabinopyranosyl)oxy]-23-hydroxyolean-12-en-28-oic acid O-alpha-L-rhamnopyranosyl-(1-->4)-O-beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranosyl ester, 3beta-[(O-beta-D-glucuronopyranosyl-(1-->3)-O-[alpha-L-rhamnopyranosyl-(1-->2)]-alpha-L-arabinopyranosyl)oxy]-23-hydroxyolean-12-en-28-oic acid O-alpha-L-rhamnopyranosyl-(1-->4)-O-beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranosyl ester, 3beta-[(O-beta-D-glucopyranosyl-(1-->3)-O-[alpha-L-rhamnopyranosyl-(1-->2)]-alpha-L-arabinopyranosyl)oxy]-29-hydroxyolean-12-en-28-oic acid, and 3beta-[(O-beta-D-glucopyranosyl-(1-->3)-O-[alpha-L-rhamnopyranosyl-(1-->2)]-alpha-L-arabinopyranosyl)oxy]-23,29-dihydroxyolean-12-en-28-oic acid, respectively. The main triterpene glycosides contained in the stems of A. quinata were found to have two sugar units at C-3 and C-28 of the aglycone in this study, whereas those of Akebia trifoliate were reported to possess one sugar unit at C-28 of the aglycone. It may be possible to distinguish between A. quinata and A. trifoliate chemically by comparing their triterpene glycoside constituents. [Abstract/Link to Full Text]

Rukachaisirikul V, Sommart U, Phongpaichit S, Hutadilok-Towatana N, Rungjindamai N, Sakayaroj J
Metabolites from the xylariaceous fungus PSU-A80.
Chem Pharm Bull (Tokyo). 2007 Sep;55(9):1316-8.
One hypoxylonol, xylarenol (1), one hexadienoic acid, xylarenoic acid (2), and one tetralone, xylarenone (3), were isolated from the xylariaceous fungus PSU-A80 together with ten known compounds. The structures were established by analysis of spectroscopic data. 8-Methoxy-1-naphthol, one of the known metabolites, displayed good radical scavenging potency with an IC(50) value of 30 microg/ml. [Abstract/Link to Full Text]

Yoshikawa M, Wang T, Morikawa T, Xie H, Matsuda H
Bioactive constituents from chinese natural medicines. XXIV. Hypoglycemic effects of Sinocrassula indica in sugar-loaded rats and genetically diabetic KK-A(y) mice and structures of new acylated flavonol glycosides, sinocrassosides A(1), A(2), B(1), and B(2).
Chem Pharm Bull (Tokyo). 2007 Sep;55(9):1308-15.
The methanolic extract from the whole plant of Sinocrassula indica (Crassulaceae) was found to inhibit the increase in serum glucose levels in oral administration of sucrose and glucose in rats at a dose of 250 mg/kg (p.o.). However, the extract did not inhibit the increase in serum glucose levels after intraperitoneal administration of glucose in these animals but did partly inhibit the gastric emptying. On the other hand, this extract significantly inhibited the increase in serum glucose levels after administration for 2 weeks in KK-A(y) mice, a genetically type II diabetic mice, at a dose of 250 mg/kg/d (p.o.) without significant changes of the weights of body, liver, and visceral fat. From the extract, four new acylated flavonol glycosides, sinocrassosides A(1), A(2), B(1), and B(2), were isolated together with 11 flavonoids and 2 megastigmanes. The absolute stereostructures of the four new compounds were elucidated on the basis of chemical and physicochemical evidence. [Abstract/Link to Full Text]

Xu GQ, Guo P, Zhang C, He QJ, Yang B, Hu YZ
Synthesis and cytotoxicity of indolopyrrolemaleimides.
Chem Pharm Bull (Tokyo). 2007 Sep;55(9):1302-7.
A series of indolopyrrolemaleimides have been synthesized and evaluated for their cytotoxicity in vitro against human leukemia cell line and four human solid cancer cell lines. Some of the compounds showed high or mediate activity against the lines. 6dc is the most promising compound among them. The inhibition toward topoisomerase I was also studied. [Abstract/Link to Full Text]

Sung PJ, Chuang LF, Kuo J, Chen JJ, Fan TY, Li JJ, Fang LS, Wang WH
Rumphellolides A-F, six new caryophyllane-related derivatives from the Formosan Gorgonian coral Rumphella antipathies.
Chem Pharm Bull (Tokyo). 2007 Sep;55(9):1296-301.
Six new caryophyllane-related natural products, including two carboxylated sesquiterpenoids, rumphellolides A (1) and B (2), and four norsesquiterpene alcohols, rumphellolides C-F (3-6), were isolated from the Formosan gorgonian coral Rumphella antipathies. The structures of the above new natural products were established on the basis of extensive spectral data analysis. Rumphellolides A (1), D (4), E (5), and F (6) showed weak antibacterial activity. [Abstract/Link to Full Text]

Sun H, Lv H, Zhang Y, Wang X, Bi K, Cao H
Pharmacokinetics of isofraxidin in rat plasma after oral administration of the extract of Acanthopanax senticosus using HPLC with solid phase extraction method.
Chem Pharm Bull (Tokyo). 2007 Sep;55(9):1291-5.
High-performance liquid chromatography coupled with solid phase extraction method was developed for determination of isofraxidin in rat plasma after oral administration of Acanthopanax senticosus extract (ASE), and pharmacokinetic parameters of isofraxidin either in ASE or pure compound were measured. The HPLC analysis was performed on a Dikma Diamonsil RP(18) column (4.6 mm x 150 mm, 5 microm) with the isocratic elution of solvent A (acetonitrile) and solvent B (0.1% aqueous phosphoric acid, v/v) (A : B = 22 : 78) and the detection wavelength was set at 343 nm. The calibration curve was linear over the range of 0.156-15.625 microg/ml. The limit of detection was 60 ng/ml. The intra-day precision was 5.8%, and the inter-day precision was 6.0%. The recovery was 87.30+/-1.73%. When the dosage of ASE is equal to pure compound caculated by the amount of isofraxidin, it has been found to have two maximum concentrations in plasma while the pure compound only showed one peak in the plasma concentration-time curve. The determined content of isofraxidin in plasma after oral administration of ASE is the total contents of free isofraxidin and its precursors in ASE in vitro. The pharmacokinetic characteristics of ASE showed the priority of the extract and the properities of traditional Chinese medicine. [Abstract/Link to Full Text]

Zhou GX, Jiang RW, Cheng Y, Ye WC, Shi JG, Gong NB, Lu Y
Daphnogirins A and B, two biflavones from Daphne giraldii.
Chem Pharm Bull (Tokyo). 2007 Sep;55(9):1287-90.
Two new biflavonoids, daphnogirins A (1) and B (2), were obtained from the roots of Daphne giraldii. Their structures were established on the basis of the spectral data and X-ray diffraction data of the co-crystal of 1 and 2. Daphnogrins A and B have the same configuration at C-1 and opposite configurations at C-16 and C-17. Oxygen radical scavenging assay has indicated that they are of significant antioxidative activity. [Abstract/Link to Full Text]

Yakura T, Sato S, Yoshimoto Y
Enantioselective synthesis of pachastrissamine (jaspin B) using dirhodium(II)-catalyzed C-H amination and asymmetric dihydroxylation as key steps.
Chem Pharm Bull (Tokyo). 2007 Aug;55(8):1284-6.
Enantioselective total synthesis of anhydrophytosphingosine pachastrissamine (jaspin B) was achieved using Sharpless asymmetric dihydroxylation and dirhodium(II)-catalyzed C-H amination as key steps. [Abstract/Link to Full Text]

Lei C, Huang SX, Chen JJ, Pu JX, Yang LB, Zhao Y, Liu JP, Gao XM, Xiao WL, Sun HD
Lignans from Schisandra propinqua var. propinqua.
Chem Pharm Bull (Tokyo). 2007 Aug;55(8):1281-3.
Two new dibenzocyclooctadiene lignans angeloyl-(+)-gomisin K(3) (1) and methylisogomisin O (2), together with six known ones, isogomisin O, angeloylisogomisin O, gomisin O, angeloygomisin O, benzoylgomisin O, epigomisin O, and four 1,4-bis(phenyl)-2,3-dimethylbutane type lignans, pregomisin, meso-dihydroguaiaretic acid, isoanwulignan, and sphenanlignan were isolated from the aerial parts of Schisandra propinqua var. propinqua. The structures of 1 and 2 were elucidated by spectroscopic methods including extensive 1D- and 2D-NMR techniques. [Abstract/Link to Full Text]

Kanchanapoom T, Otsuka H, Ruchirawat S
Megastigmane glucosides from Equisetum debile and E. diffusum.
Chem Pharm Bull (Tokyo). 2007 Aug;55(8):1277-80.
A new megastigmane diglucoside, (3S,5R,6S,7E,9S)-megastigman-7-ene-5,6-epoxy-3,9-diol 3,9-O-beta-D-diglucopyranoside (3), was isolated from the aerial portion of Equisetum debile along with macarangioside D (debiloside A), sammangaoside A, (6R,9S)-3-oxo-alpha-ionol 9-O-beta-D-glucopyranoside, debiloside B, kaempferol 3-O-sophoroside, kaempferol 3,7-O-beta-D-diglucopyranoside, kaempferol 3-O-sophoroside-7-O-beta-D-glucopyranoside, phenylethyl O-beta-D-glucopyranoside, (Z)-3-hexenyl O-beta-D-glucopyranoside, (7S,8R)-dehydrodiconiferyl 4-O-beta-D-glucopyranoside, and L-tryptophan. The absolute configuration at C-6 of the original structure of debilo-side A was revised to 6R-configuration, and was identical with macarangioside D (1). From the aerial portion of E. diffusum, four compounds, sammangaoside A, kaempferol 3-O-sophoroside and L-tryptophan and (3S,5R,6S,7E,9S)-megastigman-7-ene-5,6-epoxy-3,9-diol 3-O-beta-D-glucopyranoside were identified. The spectroscopic data of (3S,5R,6S,7E,9S)-megastigman-7-ene-5,6-epoxy-3,9-diol 3-O-beta-D-glucopyranoside (13) were found to be identical with corchoionoside A (9R-isomeric compound). The structure of corchoionoside A was also discussed. Structure determinations were based on physical data and spectroscopic evidence. [Abstract/Link to Full Text]

Das B, Chowdhury N, Damodar K, Banerjee J
A mild and efficient stereoselective synthesis of (Z)- and (E)-allyl sulfides and potent antifungal agent, (Z)-3-(4-methoxybenzylidene)thiochroman-4-one from Morita-Baylis-Hillman acetates.
Chem Pharm Bull (Tokyo). 2007 Aug;55(8):1274-6.
A facile stereoselective synthesis of (Z)- and (E)-allyl sulfides has been accomplished from Morita-Baylis-Hillman acetates in one-pot by treatment with benzene thiol in the presence of catalytic amounts of 15% aqueous NaOH and TBAI in DMSO at room temperature. The method has been applied for the synthesis of (Z)-3-(4-methoxybenzylidene)thiochroman-4-one, a potent antifungal compound. [Abstract/Link to Full Text]

Han AR, Kim JB, Lee J, Nam JW, Lee IS, Shim CK, Lee KT, Seo EK
A new flavanone glycoside from the dried immature fruits of Poncirus trifoliata.
Chem Pharm Bull (Tokyo). 2007 Aug;55(8):1270-3.
A new flavanone glycoside, (2R)-5-hydroxy-4'-methoxyflavanone-7-O-{beta-glucopyranosyl-(1-->2)-beta-glucopyranoside} (1), was isolated from the EtOAc extract of dried immature fruit of Poncirus trifoliata, together with three known compounds, (2S)-poncirin (2), (2S)-naringin (3), and (2S)-poncirenin (4). The structure of compound 1 was elucidated by spectroscopic data analysis, including 1D and 2D NMR experiments. Among the isolates, compound 2 exhibited considerable inhibitory activity against lipopolysaccharide (LPS)-induced prostaglandin E(2) (PGE(2)) and interleukin-6 (IL-6) production, and mRNA expression in RAW 264.7 murine macrophage cells. [Abstract/Link to Full Text]

Zhang L, Yang Z, Tian JK
Two new indolopyridoquinazoline alkaloidal glycosides from Ranunculus ternatus.
Chem Pharm Bull (Tokyo). 2007 Aug;55(8):1267-9.
Two new indolopyridoquinazoline alkaloidal glycosides, 11-O-beta-D-glucopyranosyl rutaecarpine (ternatoside C) and 11-O-alpha-L-rhamnosyl-(1-->6)-beta-D-glucopyranosyl rutaecarpine (ternatoside D) were isolated from the roots of Ranunculus ternatus. Their structures were determined on the basis of spectroscopic and chemical methods. [Abstract/Link to Full Text]

Behery FA, Naeem ZE, Maatooq GT, Amer MM, Wen ZH, Sheu JH, Ahmed AF
Phenanthrenoids from Juncus acutus L., new natural lipopolysaccharide-inducible nitric oxide synthase inhibitors.
Chem Pharm Bull (Tokyo). 2007 Aug;55(8):1264-6.
The novel natural product juncutol (1), 1,4,7-trimethyl-8,9-dihydro-4H-cyclopenta[def]phenanthrene-2,6-diol, along with the three related metabolites juncusol (2), dehydrojuncusol (3), and 6-hydroxymethyl-1-methyl-5-vinyl-9,10-dihydrophenanthrene-2-ol (4), were isolated from the rhizomes of Juncus acutus L. (Juncaceae) growing in Egypt. The structural identity of 1 was determined on the basis of spectroscopic analyses, including 2D NMR spectroscopy. The inhibitory effect of these natural products on the expression of inducible nitric oxide synthase (iNOS) in lipopolysaccharide-stimulated RAW264.7 macrophage cells was determined for the first time. The unprecedented symmetrical compound juncutol (1) was found to be the most potent inhibitor against the induction of the proinflammatory iNOS protein. [Abstract/Link to Full Text]

Zhang X, Wang Y, Wang J, Wang Y, Li S
Effect of pore former on the properties of casted film prepared from blends of Eudragit NE 30 D and Eudragit L 30 D-55.
Chem Pharm Bull (Tokyo). 2007 Aug;55(8):1261-3.
The casted films of aqueous dispersions of Eudragit NE30 D and Eudragit L30 D-55 containing pore former were prepared. The study investigated the influence of pore former on basic model drug clarithromycin release, water uptake and water vapor permeability from casted film prepared from the blends of neutral polymer dispersion of Eudragit NE30 D and enteric polymer dispersion of Eudragit L30 D-55. This study was concluded that pore former hydroxypropyl methyl cellulose, lactose, polyethylene glycol (PEG) and polyvinyl pyrrolidon (PVP) was released at the beginning of the release process, the rate and extent of water uptake of the polymeric films were much higher in phosphate buffer pH 6.8 than in pH 5.0 and the concentration of pore former have a significant influence on the permeability to water vapour. [Abstract/Link to Full Text]

Xie HG, Chen H, Cao B, Zhang HW, Zou ZM
Cytotoxic germacranolide sesquiterpene from Inula cappa.
Chem Pharm Bull (Tokyo). 2007 Aug;55(8):1258-60.
A new germacranolide, inulacappolide (1), was isolated from the EtOH extract of the whole plant of Inula cappa along with 16 known compounds. The structure of inulacappolide was a rare 1(10)-saturated type of germacran-6,12-olide, identified as 2alpha-acetoxy-3beta-hydroxy-9beta-angeloyloxygermacra-4-en-6alpha,12-olide by spectral analysis (IR, HR-ESI/MS, (1)H-NMR, (13)C-NMR, HMQC, HMBC, NOESY). In vitro, it showed antiproliferative effects against human cervical cancer HeLa, human leukemia K562 and human nasopharyngeal carcinoma KB cell lines with IC(50) values of 1.2 microM, 3.8 microM and 5.3 microM, respectively. [Abstract/Link to Full Text]

Ravi V, Ramu E, Vijay K, Rao AS
Zn-proline catalyzed selective synthesis of 1,2-disubstituted benzimidazoles in water.
Chem Pharm Bull (Tokyo). 2007 Aug;55(8):1254-7.
Zn-proline (5 mol%) performs as a novel water-soluble and recyclable Lewis acid catalyst for the selective synthesis of 1,2-disubstituted benzimidazoles from wide range of substituted o-phenylenediamines and aldehydes in moderate to excellent isolated yields (42-92%) using water as solvent at ambient temperature. [Abstract/Link to Full Text]

Harusawa S, Kawamura M, Araki L, Taniguchi R, Yoneyama H, Sakamoto Y, Kaneko N, Nakao Y, Hatano K, Fujita T, Yamamoto R, Kurihara T, Yamatodani A
Synthesis of novel 4(5)-(5-aminotetrahydropyran-2-yl)imidazole derivatives and their in vivo release of neuronal histamine measured by brain microdialysis.
Chem Pharm Bull (Tokyo). 2007 Aug;55(8):1245-53.
The (2R,5S)-trans- and (2S,5S)-cis-stereoisomers 1a and 1b of 4(5)-(5-aminotetrahydropyran-2-yl)imidazole, which have two chiral centers and adopt a stable chair conformation, were synthesized via cyclization of diol intermediates 7 using L-glutamine as the starting material. Their enantiomers, (2S,5R)-trans-1c and (2R,5R)-cis-1d, were synthesized by the same methodology from D-glutamine. Stereo isomers 1a-d were converted into cyanoguanidines 11a-d, and into N-isopropyl and N-3,3-dimethylbutyl derivatives 12a-d and 13a-d, respectively. The results of in vivo brain microdialysis of the derivatives apparently indicated that only (2S,5R)-isomers increased the release of neuronal histamine. Among the many (2S,5R)-N-alkyl derivatives, 13c (OUP-133) and 18 (OUP-153) increased histamine release to 180-190% and 180-200% of basal levels, respectively, and were found to be novel histamine H(3) antagonists. [Abstract/Link to Full Text]

Kuroda M, Yokosuka A, Kobayashi R, Jitsuno M, Kando H, Nosaka K, Ishii H, Yamori T, Mimaki Y
Sesquiterpenoids and flavonoids from the aerial parts of Tithonia diversifolia and their cytotoxic activity.
Chem Pharm Bull (Tokyo). 2007 Aug;55(8):1240-4.
Cytotoxicity-guided fractionation of the 80% EtOH extract of Tithonia diversifolia has resulted in the isolation of twelve sesquiterpenoids (1-12), including three new ones (4, 10, 12), and three known flavonoids (13-15). The structures of the new compounds were determined by analysis of their spectroscopic data. The isolated compounds showed cytotoxic activity against HL-60 leukemia cells with IC(50) values ranging from 0.13 to 13.0 microM, when etoposide used as a positive control gave an IC(50) value of 0.43 microM. The cancer growth inhibitory property of 9, the main cytotoxic compound in T. diversifolia, was examined using a disease-oriented panel composed of 39 human cancer cell lines in the Japanese Foundation for Cancer Research. [Abstract/Link to Full Text]

Nozawa D, Okubo T, Chaki S, Okuyama S, Nakazato A
Identification of arginine analogues as antagonists and agonists for the melanocortin-4 receptor.
Chem Pharm Bull (Tokyo). 2007 Aug;55(8):1232-9.
In the present study, conducted to explore potent and small molecular melanocortin-4 (MC4) receptor ligands, we found that tripeptide 3a, containing a D-Phe-Arg-2-Nal (Nal; naphthylalanine) sequence, exhibited a moderate affinity for the MC4 receptor. Structural optimization led to the identification of a compound with a high affinity for the MC4 receptor, namely, tripeptide 3e, which showed a 70-fold higher affinity for the MC4 receptor than the lead compound 3a. Moreover, in an effort to further reduce the peptidic characters of tripeptide 3e, we found that dipeptide 3g exhibited a relatively high affinity for the MC4 receptor. Furthermore, in these analogues, the substituted position (1' vs. 2') of the naphthyl ring of Nal residue at position 7 was found to be important for the differentiation of agonist and antagonist activity. The synthesis and structure-activity relationships of the arginine analogues as MC4 receptor ligands were described in this paper. [Abstract/Link to Full Text]

Aso Y, Yoshioka S, Miyazaki T, Kawanishi T, Tanaka K, Kitamura S, Takakura A, Hayashi T, Muranushi N
Miscibility of nifedipine and hydrophilic polymers as measured by (1)H-NMR spin-lattice relaxation.
Chem Pharm Bull (Tokyo). 2007 Aug;55(8):1227-31.
The miscibility of a drug with excipients in solid dispersions is considered to be one of the most important factors for preparation of stable amorphous solid dispersions. The purpose of the present study was to elucidate the feasibility of (1)H-NMR spin-lattice relaxation measurements to assess the miscibility of a drug with excipients. Solid dispersions of nifedipine with the hydrophilic polymers poly(vinylpyrrolidone) (PVP), hydroxypropylmethylcellulose (HPMC) and alpha,beta-poly(N-5-hydroxypentyl)-L-aspartamide (PHPA) with various weight ratios were prepared by spray drying, and the spin-lattice relaxation decay of the solid dispersions in a laboratory frame (T(1) decay) and in a rotating frame (T(1rho) decay) were measured. T(1rho) decay of nifedipine-PVP solid dispersions (3 : 7, 5 : 5 and 7 : 3) was describable with a mono-exponential equation, whereas T(1rho) decay of nifedipine-PHPA solid dispersions (3 : 7, 4 : 6 and 5 : 5) was describable with a bi-exponential equation. Because a mono-exponential T(1rho) decay indicates that the domain sizes of nifedipine and polymer in solid dispersion are less than several nm, it is speculated that nifedipine is miscible with PVP but not miscible with PHPA. All the nifedipine-PVP solid dispersions studied showed a single glass transition temperature (T(g)), whereas two glass transitions were observed for the nifedipine-PHPA solid dispersion (3 : 7), thus supporting the above speculation. For nifedipine-HPMC solid dispersions (3 : 7 and 5 : 5), the miscibility of nifedipine and HPMC could not be determined by DSC measurements due to the lack of obviously evident T(g). In contrast, (1)H-NMR spin-lattice relaxation measurements showed that nifedipine and HPMC are miscible, since T(1rho) decay of the solid dispersions (3 : 7, 5 : 5 and 7 : 3) was describable with a mono-exponential equation. These results indicate that (1)H-NMR spin-lattice relaxation measurements are useful for assessing the miscibility of a drug and an excipient in solid dispersions. [Abstract/Link to Full Text]

Nakajima Y, Sato Y, Konishi T
Antioxidant small phenolic ingredients in Inonotus obliquus (persoon) Pilat (Chaga).
Chem Pharm Bull (Tokyo). 2007 Aug;55(8):1222-6.
Inonotus obliquus (persoon) Pilat (Chaga, in Russia, kabanoanatake in Japan) is a fungus having been used as a folk medicine in Russia and said to have many health beneficial functions such as immune modulating and anti-cancer activities. In the present study, the antioxidant activity of hot water extract (decoction) of Chaga was precisely compared with those of other medicinal fungi (Agaricus blazei Mycelia, Ganoderma lucidum and Phellinus linteus) showing Chaga had the strongest antioxidant activity among fungi examined in terms of both superoxide and hydroxyl radicals scavenging activities. Further determination of the antioxidant potential of isolated fruiting body (brown part) and Sclerotium (black part) revealed the 80% MeOH extract of fruiting body had the highest potential as high as that of Chaga decoction. Finally, seven antioxidant components were isolated and purified from the 80% MeOH extract of Chaga fruiting body, and their chemical structures were determined as small phenolics as follows: 4-hydroxy-3,5-dimethoxy benzoic acid 2-hydroxy-1-hydroxymethyl ethyl ester (BAEE), protocatechic acid (PCA), caffeic acid (CA), 3,4-dihybenzaladehyde (DB), 2,5-dihydroxyterephtalic acid (DTA), syringic acid (SA) and 3,4-dihydroxybenzalacetone (DBL). Notably, BAEE was assigned as the new compound firstly identified from the natural source in the present study. [Abstract/Link to Full Text]

Dai SJ, Wang GF, Chen M, Liu K, Shen L
Five new neo-clerodane diterpenoid alkaloids from Scutellaria barbata with cytotoxic activities.
Chem Pharm Bull (Tokyo). 2007 Aug;55(8):1218-21.
Five new neo-clerodane diterpenoid alkaloids, named scutebarbatine G (1), 6,7-di-O-nicotinoylscutebarbatine G (2), 6-O-nicotinoyl-7-O-acetylscutebarbatine G (3), scutebarbatine H (4) and 7-O-nicotinoylscutebarbatine H (5) were isolated from the whole plant of Scutellaria barbata D. DON. Their structures were elucidated by spectroscopic methods including extensive 1D and 2D NMR analyses. In vitro, compounds 1-5 showed significant cytotoxic activities against three human cancer lines, namely, HONE-1 nasopharyngeal, KB oral epidermoid carcinoma, and HT29 colorectal carcinoma cells, and gave IC(50) values in the range 3.4-8.5 microM. [Abstract/Link to Full Text]

Tobiishi S, Sasada T, Nojiri Y, Yamamoto F, Mukai T, Ishiwata K, Maeda M
Methoxy- and fluorine-substituted analogs of O-1302: synthesis and in vitro binding affinity for the CB1 cannabinoid receptor.
Chem Pharm Bull (Tokyo). 2007 Aug;55(8):1213-7.
Methoxy and fluorine analogs substituted on the terminal carbon of the pentyl chain of N-(piperidinyl)-1-(2,4-dichlorophenyl)-4-methyl-5-(4-pentylphenyl)-1H-pyrazole-3-carboxamide (O-1302) were synthesized in a multi-step process from 5-phenyl-1-pentanol, which was based on the 1,5-diarylpyrazole core template of N-(piperidinyl)-5-(4-chlorophenyl)-1-(2,4-dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide (SR141716) through condensation of the respective amine with pyrazole carboxylic acid, in order to develop tracers for medical imaging. Their potency for inhibiting the binding of the CB1 antagonist [(3)H]SR141716 was evaluated with the aim of developing positron emission tomography (PET) ligands for the cerebral cannabinoid CB1 receptor. These analogs bearing a piperidinyl carboxamide at the C(3) of the pyrazole ring exhibited affinities comparable to those of the CB1 reference antagonist SR141716, which warrants further investigation using the radiolabeled form for biological imaging studies. A morpholine ring substituted at the C(3) of the pyrazole ring resulted in a reduction of the CB1 affinity. [Abstract/Link to Full Text]

Tahara K, Yano Y, Kanagawa K, Abe Y, Yamada J, Iijima S, Mochizuki M, Nishikawa T
Successful preparation of metabolite of troglitazone by in-flow electrochemical reaction on coulometric electrode.
Chem Pharm Bull (Tokyo). 2007 Aug;55(8):1207-12.
A simple, rapid and efficient system utilizing a coulometric electrode was developed for the preparation of drug metabolites. Trace amounts of reactants are usually generated in electrochemical reactions, which are not suitable for the sufficient preparation of products to obtain NMR and other spectral data for chemical structure confirmation or to obtain data from pharmacological activity screening tests of products. In the developed system, called the "in-flow electrochemical reaction system," a drug, troglitazone, was dissolved in a volatile flow solvent, and pumped into a coulometric electrode under optimized conditions, and the effluent was evaporated. Without any further purification, milligram amounts of a pure oxidation product of troglitazone could be obtained within several hours. The amount obtained was enough for (1)H- and (13)C-NMR analysis by which the structure could be confirmed and was found to be identical to one of the metabolites of troglitazone detected in human plasma. This system will be useful to prepare standard compounds of the required amount for pharmacokinetic study and for toxicokinetic study. [Abstract/Link to Full Text]

Narazaki R, Sanghvi R, Yalkowsky SH
Estimation of drug precipitation upon dilution of pH-cosolvent solubilized formulations.
Chem Pharm Bull (Tokyo). 2007 Aug;55(8):1203-6.
This manuscript is a study of precipitation of insoluble drug upon dilution from the pH-cosolvent solubilized formulation with simulated blood fluid. An equation is developed to estimate drug precipitation upon dilution of combined pH-cosolvent solubilized formulations. This is an extension of a previous equation used for the estimation of drug precipitation from simple pH controlled formulations. The proposed equation considers the effect of the cosolvent and its concentration on the pK(a) of the drug as well as all buffering species. According to the proposed equation and our experimental data, the addition of cosolvent on the pH solubilized formulation could increase total drug solubility in the formulation. However, the solubility after dilution became lower than the 0% ethanol formulation because of the change in both drug and buffer pK(a) values. Since this equation is based on the equilibrium condition, it is the worst case scenario for precipitation. This equation provides useful information regarding the feasibility of the successful use of pH-cosolvent combinations in drug formulation. [Abstract/Link to Full Text]

Davarani SS, Shamsipur M, Nematollahi D, Ramyar S, Masoumi L
An environmentally friendly electrochemical method for synthesis of benzofuranoquinone derivatives.
Chem Pharm Bull (Tokyo). 2007 Aug;55(8):1198-202.
Electrochemical oxidation of catechols (1a-c) has been studied in the presence of 2-hydroxy-1,4-naphtoquinone (3b) in aqueous solutions, using cyclic voltammetry and controlled-potential coulometry. The results indicated that the electrochemically generated o-benzoquinones (2a-c) participate in Michael addition reaction with 3b to the corresponding benzofuranoquinones (8a-c, 10a-c). The electrochemical synthesis of these compounds has been successfully preformed at a carbon rod electrode with good yields using an environmentally friendly method. [Abstract/Link to Full Text]

Qin J, Chen D, Hu H, Cui Q, Qiao M, Chen B
Surface modification of RGD-liposomes for selective drug delivery to monocytes/neutrophils in brain.
Chem Pharm Bull (Tokyo). 2007 Aug;55(8):1192-7.
In the present study, RGD peptide was coupled with ferulic acid (FA) liposomes for binding to monocytes and neutrophils in peripheral blood for brain targeting in response to leukocyte recruitment. Cholesterol (Ch) was esterified with succinic anhydride to introduce a carboxylic end group (Ch-COOH). Soybean phosphatidylcholine, cholesterol and Ch-COOH were in a molar ratio of 1 : 0.23 : 0.05. FA was loaded into liposomes with 80.2+/-5.2% entrapment efficiency (EE) using a calcium acetate gradient method since it was difficult to load FA by other methods. RGD peptide was a novel compound coupled with Ch-COOH via carbodiimide and N-hydroxysulfosuccinimide. The results of the in vitro flow cytometric study showed that RGD conjugation liposomes (RGD-liposomes) could bind to monocytes/neutrophils efficiently. The rats were subjected to intrastriatal microinjections of 100 microl of human recombinant IL-1beta to produce brain inflammation and subsequently sacrificed after 15, 30, 60 and 120 min of administration of three formulations (FA solution, FA liposome, RGD-coated FA liposome). The body distribution results showed that RGD-liposomes could be directed to the target site, i.e. the brain, by cell selectivity in case of an inflammatory response. For RGD coated liposomes, the concentration of FA in brain was 6-fold higher than that of FA solution and 3-fold higher than that of uncoated liposomes. MTT assay and flow cytometry were used in the pharmacodynamic studies where it was found that FA liposomes exhibited greater antioxidant activity to FA solution on U937 cell. [Abstract/Link to Full Text]

Ninomiya K, Morikawa T, Zhang Y, Nakamura S, Matsuda H, Muraoka O, Yoshikawa M
Bioactive constituents from Chinese natural medicines. XXIII. Absolute structures of new megastigmane glycosides, sedumosides A(4), A(5), A(6), H, and I, and hepatoprotective megastigmanes from Sedum sarmentosum.
Chem Pharm Bull (Tokyo). 2007 Aug;55(8):1185-91.
The methanol-eluted fraction of the hot water extract from the whole plant of Sedum sarmentosum (Crassulaceae) was found to show hepatoprotective effect on D-galactosamine-induced cytotoxicity in primary cultured mouse hepatocytes. From the active fraction, five new megastigmane glycosides, sedumosides A(4), A(5), A(6), H, and I, were isolated together with 22 megastigmane constituents. Their absolute stereostructures were elucidated on the basis of chemical and physicochemical evidence. Among them, sedumoside F(1) (IC(50)=47 microM), (3S,5R,6S,9R)-megastigmane-3,9-diol (61 microM), and myrsinionosides A (52 microM) and D (62 microM) were found to show the strong hepatoprotective activity. [Abstract/Link to Full Text]

Recent Articles in The Journal of Experimental Medicine

Yang CS, Lee DS, Song CH, An SJ, Li S, Kim JM, Kim CS, Yoo DG, Jeon BH, Yang HY, Lee TH, Lee ZW, El-Benna J, Yu DY, Jo EK
Roles of peroxiredoxin II in the regulation of proinflammatory responses to LPS and protection against endotoxin-induced lethal shock.
J Exp Med. 2007 Mar 19;204(3):583-94.
Mammalian 2-Cys peroxiredoxin II (Prx II) is a cellular peroxidase that eliminates endogenous H(2)O(2). The involvement of Prx II in the regulation of lipopolysaccharide (LPS) signaling is poorly understood. In this report, we show that LPS induces substantially enhanced inflammatory events, which include the signaling molecules nuclear factor kappaB and mitogen-activated protein kinase (MAPK), in Prx II-deficient macrophages. This effect of LPS was mediated by the robust up-regulation of the reactive oxygen species (ROS)-generating nicotinamide adenine dinucleotide phosphate (NADPH) oxidases and the phosphorylation of p47(phox). Furthermore, challenge with LPS induced greater sensitivity to LPS-induced lethal shock in Prx II-deficient mice than in wild-type mice. Intravenous injection of Prx II-deficient mice with the adenovirus-encoding Prx II gene significantly rescued mice from LPS-induced lethal shock as compared with the injection of a control virus. The administration of catalase mimicked the reversal effects of Prx II on LPS-induced inflammatory responses in Prx II-deficient cells, which suggests that intracellular H(2)O(2) is attributable, at least in part, to the enhanced sensitivity to LPS. These results indicate that Prx II is an essential negative regulator of LPS-induced inflammatory signaling through modulation of ROS synthesis via NADPH oxidase activities and, therefore, is crucial for the prevention of excessive host responses to microbial products. [Abstract/Link to Full Text]

Zhu J, Koelle DM, Cao J, Vazquez J, Huang ML, Hladik F, Wald A, Corey L
Virus-specific CD8+ T cells accumulate near sensory nerve endings in genital skin during subclinical HSV-2 reactivation.
J Exp Med. 2007 Mar 19;204(3):595-603.
Cytotoxic CD8(+) T cells play a critical role in controlling herpes simplex virus (HSV) infection and reactivation. However, little is known about the spatiotemporal dynamics of CD8(+) T cells during HSV lesion evolution or about their involvement in immune surveillance after lesion resolution. Using quantum dot-conjugated peptide-major histocompatibility complex multimers, we investigated the in vivo localization of HSV-2-specific CD8(+) T cells in sequential biopsies of human genital skin during acute, resolving, and healed stages of HSV-2 reactivation. Our studies revealed that functionally active CD8(+) T cells selectively infiltrated to the site of viral reactivation. After lesion healing in concert with complete reepithelialization and loss of HSV DNA from skin biopsies, HSV-2-specific CD8(+) T cells persisted for more than two months at the dermal-epidermal junction, adjacent to peripheral nerve endings. In two out of the six sequentially studied individuals, HSV-2 DNA reappeared in clinically and histologically normal-appearing skin. Detection of viral DNA was accompanied by increased numbers of both HSV-specific and total CD8(+) T cells in the dermis. These findings indicate that the frequency and clinical course of HSV-2 reactivation in humans is influenced by virus-specific CD8(+) T cells that persist in peripheral mucosa and genital skin after resolution of herpes lesions. [Abstract/Link to Full Text]

Nombela-Arrieta C, Mempel TR, Soriano SF, Mazo I, Wymann MP, Hirsch E, Mart�nez-A C, Fukui Y, von Andrian UH, Stein JV
A central role for DOCK2 during interstitial lymphocyte motility and sphingosine-1-phosphate-mediated egress.
J Exp Med. 2007 Mar 19;204(3):497-510.
Recent observations using multiphoton intravital microscopy (MP-IVM) have uncovered an unexpectedly high lymphocyte motility within peripheral lymph nodes (PLNs). Lymphocyte-expressed intracellular signaling molecules governing interstitial movement remain largely unknown. Here, we used MP-IVM of murine PLNs to examine interstitial motility of lymphocytes lacking the Rac guanine exchange factor DOCK2 and phosphoinositide-3-kinase (PI3K)gamma, signaling molecules that act downstream of G protein-coupled receptors, including chemokine receptors (CKRs). T and B cells lacking DOCK2 alone or DOCK2 and PI3Kgamma displayed markedly reduced motility inside T cell area and B cell follicle, respectively. Lack of PI3Kgamma alone had no effect on migration velocity but resulted in increased turning angles of T cells. As lymphocyte egress from PLNs requires the sphingosine-1-phosphate (S1P) receptor 1, a G(alphai) protein-coupled receptor similar to CKR, we further analyzed whether DOCK2 and PI3Kgamma contributed to S1P-triggered signaling events. S1P-induced cell migration was significantly reduced in T and B cells lacking DOCK2, whereas T cell-expressed PI3Kgamma contributed to F-actin polymerization and protein kinase B phosphorylation but not migration. These findings correlated with delayed lymphocyte egress from PLNs in the absence of DOCK2 but not PI3Kgamma, and a markedly reduced cell motility of DOCK2-deficient T cells in close proximity to efferent lymphatic vessels. In summary, our data support a central role for DOCK2, and to a lesser extent T cell-expressed PI3Kgamma, for signal transduction during interstitial lymphocyte migration and S1P-mediated egress. [Abstract/Link to Full Text]

Worbs T, Mempel TR, B�lter J, von Andrian UH, F�rster R
CCR7 ligands stimulate the intranodal motility of T lymphocytes in vivo.
J Exp Med. 2007 Mar 19;204(3):489-95.
In contrast to lymphocyte homing, little is known about molecular cues controlling the motility of lymphocytes within lymphoid organs. Applying intravital two-photon microscopy, we demonstrate that chemokine receptor CCR7 signaling enhances the intranodal motility of CD4(+) T cells. Compared to wild-type (WT) cells, the average velocity and mean motility coefficient of adoptively transferred CCR7-deficient CD4(+) T lymphocytes in T cell areas of WT recipients were reduced by 33 and 55%, respectively. Both parameters were comparably reduced for WT T lymphocytes migrating in T cell areas of plt/plt mice lacking CCR7 ligands. Importantly, systemic application of the CCR7 ligand CCL21 was sufficient to rescue the motility of WT T lymphocytes inside T cell areas of plt/plt recipients. Comparing the movement behavior of T cells in subcapsular areas that are devoid of detectable amounts of CCR7 ligands even in WT mice, we failed to reveal any differences between WT and plt/plt recipients. Furthermore, in both WT and plt/plt recipients, highly motile T cells rapidly accumulated in the subcapsular region after subcutaneous injection of the CCR7 ligand CCL19. Collectively, these data identify CCR7 and its ligands as important chemokinetic factors stimulating the basal motility of CD4(+) T cells inside lymph nodes in vivo. [Abstract/Link to Full Text]

Le Floc'h A, Jalil A, Vergnon I, Le Maux Chansac B, Lazar V, Bismuth G, Chouaib S, Mami-Chouaib F
Alpha E beta 7 integrin interaction with E-cadherin promotes antitumor CTL activity by triggering lytic granule polarization and exocytosis.
J Exp Med. 2007 Mar 19;204(3):559-70.
Various T cell adhesion molecules and their cognate receptors on target cells promote T cell receptor (TCR)-mediated cell killing. In this report, we demonstrate that the interaction of epithelial cell marker E-cadherin with integrin alpha(E)(CD103)beta(7), often expressed by tumor-infiltrating lymphocytes (TILs), plays a major role in effective tumor cell lysis. Indeed, we found that although tumor-specific CD103(+) TIL-derived cytotoxic T lymphocyte (CTL) clones are able to kill E-cadherin(+)/intercellular adhesion molecule 1(-) autologous tumor cells, CD103(-) peripheral blood lymphocyte (PBL)-derived counterparts are inefficient. This cell killing is abrogated after treatment of the TIL clones with a blocking anti-CD103 monoclonal antibody or after targeting E-cadherin in the tumor using ribonucleic acid interference. Confocal microscopy analysis also demonstrated that alpha(E)beta(7) is recruited at the immunological synapse and that its interaction with E-cadherin is required for cytolytic granule polarization and subsequent exocytosis. Moreover, we report that the CD103(-) profile, frequently observed in PBL-derived CTL clones and associated with poor cytotoxicity against the cognate tumor, is up-regulated upon TCR engagement and transforming growth factor beta1 treatment, resulting in strong potentiation of antitumor lytic function. Thus, CD8(+)/CD103(+) tumor-reactive T lymphocytes infiltrating epithelial tumors most likely play a major role in antitumor cytotoxic response through alpha(E)beta(7)-E-cadherin interactions. [Abstract/Link to Full Text]

Tellam J, Fogg MH, Rist M, Connolly G, Tscharke D, Webb N, Heslop L, Wang F, Khanna R
Influence of translation efficiency of homologous viral proteins on the endogenous presentation of CD8+ T cell epitopes.
J Exp Med. 2007 Mar 19;204(3):525-32.
A significant proportion of endogenously processed CD8(+) T cell epitopes are derived from newly synthesized proteins and rapidly degrading polypeptides (RDPs). It has been hypothesized that the generation of rapidly degrading polypeptides and CD8(+) T cell epitopes from these RDP precursors may be influenced by the efficiency of protein translation. Here we address this hypothesis by using the Epstein-Barr virus-encoded nuclear antigen 1 protein (EBNA1), with or without its internal glycine-alanine repeat sequence (EBNA1 and EBNA1DeltaGA, respectively), which display distinct differences in translation efficiency. We demonstrate that RDPs constitute a significant proportion of newly synthesized EBNA1 and EBNA1DeltaGA and that the levels of RDPs produced by each of these proteins directly correlate with the translation efficiency of either EBNA1 or EBNA1DeltaGA. As a consequence, a higher number of major histocompatibility complex-peptide complexes can be detected on the surface of cells expressing EBNA1DeltaGA, and these cells are more efficiently recognized by virus-specific cytotoxic T lymphocytes compared to the full-length EBNA1. More importantly, we also demonstrate that the endogenous processing of these CD8(+) T cell epitopes is predominantly determined by the rate at which the RDPs are generated rather than the intracellular turnover of these proteins. [Abstract/Link to Full Text]

Dooms H, Wolslegel K, Lin P, Abbas AK
Interleukin-2 enhances CD4+ T cell memory by promoting the generation of IL-7R alpha-expressing cells.
J Exp Med. 2007 Mar 19;204(3):547-57.
The common gamma chain cytokines interleukin (IL)-2 and IL-7 are important regulators of T cell homeostasis. Although IL-2 is implicated in the acute phase of the T cell response, IL-7 is important for memory T cell survival. We asked whether regulated responsiveness to these growth factors is determined by temporal expression of the cytokine-specific IL-2 receptor (R) alpha and IL-7Ralpha chains. We demonstrate that IL-2Ralpha is expressed early after priming in T cell receptor-transgenic CD4(+) T cells, whereas IL-7Ralpha expression is lost. In the later stage of the response, IL-7Ralpha is reexpressed while IL-2Ralpha expression is silenced. This reciprocal pattern of IL-2Ralpha/IL-7Ralpha expression is disturbed when CD4(+) T cells are primed in the absence of IL-2 signals. Primed IL-2(-/-) or CD25(-/-) (IL-2Ralpha(-/-)) CD4(+) T cells, despite showing normal induction of activation markers and cell division, fail to reexpress IL-7Ralpha late in the response. Because the generation of CD4(+) memory T cells is dependent on IL-7-IL-7Ralpha interactions, primed IL-2(-/-) or CD25(-/-) CD4(+) T cells develop poorly into long-lived memory cells. Retrovirus-mediated expression of IL-7Ralpha in IL-2(-/-) T cells restores their capacity for long-term survival. These results identify IL-2 as a factor regulating IL-7Ralpha expression and, consequently, memory T cell homeostasis in vivo. [Abstract/Link to Full Text]

Dougan SK, Rava P, Hussain MM, Blumberg RS
MTP regulated by an alternate promoter is essential for NKT cell development.
J Exp Med. 2007 Mar 19;204(3):533-45.
Microsomal triglyceride transfer protein (MTP), an endoplasmic reticulum lipid transfer protein critical for apolipoprotein B (apoB) secretion, regulates CD1d antigen presentation. We identified MTP variant 1 (MTPv1), a novel splice variant of mouse MTP, by polymerase chain reaction and Northern analysis in non-apoB-secreting tissues, including thymocytes and antigen-presenting cells (APCs). Edman degradation of MTPv1 isolated from transfected cells revealed three unique residues; however, recombinant MTP and MTPv1 had an equivalent protein disulfide isomerase association, subcellular localization, triglyceride transfer, phospholipid transfer, response to inhibitors, and ability to support apoB secretion. MTP and MTPv1 efficiently transferred phosphatidylethanolamine to CD1d in vitro. NKT cells fail to develop in fetal thymic organ culture (FTOC) treated with MTP antagonists. MTP-inhibited FTOCs produced negligible numbers of CD1d tetramer-positive cells and exhibited marked defects in IL-4 production upon stimulation with anti-CD3 or alpha-galactosylceramide-pulsed APCs. CD1d expression on CD4(+)CD8(+) FTOC cells was unaffected by MTP inhibition. Thus, our results demonstrate that MTPv1 in thymocytes is critical to NKT cell development. We hypothesize that, when MTP is inactive, CD1d traffics to the cell surface and presents no lipid or a lipid that is incapable of mediating NKT cell selection and/or is refractory to lysosomal editing. [Abstract/Link to Full Text]

Min-Oo G, Fortin A, Pitari G, Tam M, Stevenson MM, Gros P
Complex genetic control of susceptibility to malaria: positional cloning of the Char9 locus.
J Exp Med. 2007 Mar 19;204(3):511-24.
Mouse strains AcB55 and AcB61 are resistant to malaria by virtue of a mutation in erythrocyte pyruvate kinase (Pklr(I90N)). Linkage analysis in [AcB55 x A/J] F2 mice detected a second locus (Char9; logarithm of odds = 4.74) that regulates the blood-stage replication of Plasmodium chabaudi AS independently of Pklr. We characterized the 77 genes of the Char9 locus for tissue-specific expression, strain-specific alterations in gene expression, and polymorphic variants that are possibly associated with differential susceptibility. We identified Vnn1/Vnn3 as the likely candidates responsible for Char9. Vnn3/Vnn1 map within a conserved haplotype block and show expression levels that are strictly cis-regulated by this haplotype. The absence of Vnn messenger RNA expression and lack of pantetheinase protein activity in tissues are associated with susceptibility to malaria and are linked to a complex rearrangement in the Vnn3 promoter region. The A/J strain also carries a unique nonsense mutation that leads to a truncated protein. Vanin genes code for a pantetheinase involved in the production of cysteamine, a key regulator of host responses to inflammatory stimuli. Administration of cystamine in vivo partially corrects susceptibility to malaria in A/J mice, as measured by reduced blood parasitemia and decreased mortality. These studies suggest that pantetheinase is critical for the host response to malaria. [Abstract/Link to Full Text]

van Zelm MC, Szczepanski T, van der Burg M, van Dongen JJ
Replication history of B lymphocytes reveals homeostatic proliferation and extensive antigen-induced B cell expansion.
J Exp Med. 2007 Mar 19;204(3):645-55.
The contribution of proliferation to B lymphocyte homeostasis and antigen responses is largely unknown. We quantified the replication history of mouse and human B lymphocyte subsets by calculating the ratio between genomic coding joints and signal joints on kappa-deleting recombination excision circles (KREC) of the IGK-deleting rearrangement. This approach was validated with in vitro proliferation studies. We demonstrate that naive mature B lymphocytes, but not transitional B lymphocytes, undergo in vivo homeostatic proliferation in the absence of somatic mutations in the periphery. T cell-dependent B cell proliferation was substantially higher and showed higher frequencies of somatic hypermutation than T cell-independent responses, fitting with the robustness and high affinity of T cell-dependent antibody responses. More extensive proliferation and somatic hypermutation in antigen-experienced B lymphocytes from human adults compared to children indicated consecutive responses upon additional antigen exposures. Our combined observations unravel the contribution of proliferation to both B lymphocyte homeostasis and antigen-induced B cell expansion. We propose an important role for both processes in humoral immunity. These new insights will support the understanding of peripheral B cell regeneration after hematopoietic stem cell transplantation or B cell-directed antibody therapy, and the identification of defects in homeostatic or antigen-induced B cell proliferation in patients with common variable immunodeficiency or another antibody deficiency. [Abstract/Link to Full Text]

Wells WA
The returning tide: how China, the world's most populous country, is building a competitive research base.
J Exp Med. 2007 Feb 19;204(2):210-35.
When China turned its back on the Cultural Revolution, it aimed to build a thriving capitalist sector. It got one. Now, it wants a world-class research enterprise. How far has it progressed in the biosciences, how did it get there, and how far does it have to go? [Abstract/Link to Full Text]

Trinchieri G
Interleukin-10 production by effector T cells: Th1 cells show self control.
J Exp Med. 2007 Feb 19;204(2):239-43.
Interleukin (IL)-10 is a cytokine that modulates both innate and adaptive immunity, primarily by exerting antiinflammatory effects. IL-10 was originally thought to be produced only by T helper (Th)2 cells, but is now known to be made by a variety of cell types. During many infections, CD4(+) T cells produce both interferon (IFN)-gamma, the signature Th1 cytokine, and IL-10. New data now show that the IL-10 produced by effector Th1 cells helps limit the collateral damage caused by exaggerated inflammation. But this control may also limit the effectiveness of the immune response, resulting in a failure to fully eliminate pathogens. [Abstract/Link to Full Text]

Jenkinson SR, Intlekofer AM, Sun G, Feigenbaum L, Reiner SL, Bosselut R
Expression of the transcription factor cKrox in peripheral CD8 T cells reveals substantial postthymic plasticity in CD4-CD8 lineage differentiation.
J Exp Med. 2007 Feb 19;204(2):267-72.
Most T cells belong to either of two lineages defined by the mutually exclusive expression of CD4 and CD8 coreceptors: CD4 T cells are major histocompatibility complex (MHC) II restricted and have helper function, whereas CD8 T cells are MHC I restricted and have cytotoxic function. The divergence between these two lineages occurs during intrathymic selection and is thought to be irreversible in mature T cells. It is, however, unclear whether the CD4-CD8 differentiation of postthymic T cells retains some level of plasticity or is stably maintained by mechanisms distinct from those that set lineage choice in the thymus. To address this issue, we examined if coreceptor or effector gene expression in mature CD8 T cells remains sensitive to the zinc finger transcription factor cKrox, which promotes CD4 and inhibits CD8 differentiation when expressed in thymocytes. We show that cKrox transduction into CD8 T cells inhibits their expression of CD8 and cytotoxic effector genes and impairs their cytotoxic activity, and that it promotes expression of helper-specific genes, although not of CD4 itself. These observations reveal a persistent degree of plasticity in CD4-CD8 differentiation in mature T cells. [Abstract/Link to Full Text]

Shamshiev AT, Ampenberger F, Ernst B, Rohrer L, Marsland BJ, Kopf M
Dyslipidemia inhibits Toll-like receptor-induced activation of CD8alpha-negative dendritic cells and protective Th1 type immunity.
J Exp Med. 2007 Feb 19;204(2):441-52.
Environmental factors, including diet, play a central role in influencing the balance of normal immune homeostasis; however, many of the cellular mechanisms maintaining this balance remain to be elucidated. Using mouse models of genetic and high-fat/cholesterol diet-induced dyslipidemia, we examined the influence of dyslipidemia on T cell and dendritic cell (DC) responses in vivo and in vitro. We show that dyslipidemia inhibited Toll-like receptor (TLR)-induced production of proinflammatory cytokines, including interleukin (IL)-12, IL-6, and tumor necrosis factor-alpha, as well as up-regulation of costimulatory molecules by CD8alpha(-) DCs, but not by CD8alpha(+) DCs, in vivo. Decreased DC activation profoundly influenced T helper (Th) cell responses, leading to impaired Th1 and enhanced Th2 responses. As a consequence of this immune modulation, host resistance to Leishmania major was compromised. We found that oxidized low-density lipoprotein (oxLDL) was the key active component responsible for this effect, as it could directly uncouple TLR-mediated signaling on CD8alpha(-) myeloid DCs and inhibit NF-kappaB nuclear translocation. These results show that a dyslipidemic microenvironment can directly interfere with DC responses to pathogen-derived signals and skew the development of T cell-mediated immunity. [Abstract/Link to Full Text]

Smith AL, Ganesh L, Leung K, Jongstra-Bilen J, Jongstra J, Nabel GJ
Leukocyte-specific protein 1 interacts with DC-SIGN and mediates transport of HIV to the proteasome in dendritic cells.
J Exp Med. 2007 Feb 19;204(2):421-30.
Dendritic cells (DCs) capture and internalize human immunodeficiency virus (HIV)-1 through C-type lectins, including DC-SIGN. These cells mediate efficient infection of T cells by concentrating the delivery of virus through the infectious synapse, a process dependent on the cytoplasmic domain of DC-SIGN. Here, we identify a cellular protein that binds specifically to the cytoplasmic region of DC-SIGN and directs internalized virus to the proteasome. This cellular protein, leukocyte-specific protein 1 (LSP1), was defined biochemically by immunoprecipitation and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. LSP1 is an F-actin binding protein involved in leukocyte motility and found on the cytoplasmic surface of the plasma membrane. LSP1 interacted specifically with DC-SIGN and other C-type lectins, but not the inactive mutant DC-SIGNDelta35, which lacks a cytoplasmic domain and shows altered virus transport in DCs. LSP1 diverts HIV-1 to the proteasome. Down-regulation of LSP1 with specific small interfering RNAs in human DCs enhanced HIV-1 transfer to T cells, and bone marrow DCs from lsp1(-/-) mice also showed an increase in transfer of HIV-1(BaL) to a human T cell line. Proteasome inhibitors increased retention of viral proteins in lsp1(+/+) DCs, and substantial colocalization of virus to the proteasome was observed in wild-type compared with LSP1-deficient cells. Collectively, these data suggest that LSP1 protein facilitates virus transport into the proteasome after its interaction with DC-SIGN through its interaction with cytoskeletal proteins. [Abstract/Link to Full Text]

Maillard MH, Cotta-de-Almeida V, Takeshima F, Nguyen DD, Michetti P, Nagler C, Bhan AK, Snapper SB
The Wiskott-Aldrich syndrome protein is required for the function of CD4(+)CD25(+)Foxp3(+) regulatory T cells.
J Exp Med. 2007 Feb 19;204(2):381-91.
The Wiskott-Aldrich syndrome, a primary human immunodeficiency, results from defective expression of the hematopoietic-specific cytoskeletal regulator Wiskott-Aldrich syndrome protein (WASP). Because CD4(+)CD25(+)Foxp3(+) naturally occurring regulatory T (nTreg) cells control autoimmunity, we asked whether colitis in WASP knockout (WKO) mice is associated with aberrant development/function of nTreg cells. We show that WKO mice have decreased numbers of CD4(+)CD25(+)Foxp3(+) nTreg cells in both the thymus and peripheral lymphoid organs. Moreover, we demonstrate that WKO nTreg cells are markedly defective in both their ability to ameliorate the colitis induced by the transfer of CD45RB(hi) T cells and in functional suppression assays in vitro. Compared with wild-type (WT) nTreg cells, WKO nTreg cells show significantly impaired homing to both mucosal (mesenteric) and peripheral sites upon adoptive transfer into WT recipient mice. Suppression defects may be independent of antigen receptor-mediated actin rearrangement because both WT and WKO nTreg cells remodeled their actin cytoskeleton inefficiently upon T cell receptor stimulation. Preincubation of WKO nTreg cells with exogenous interleukin (IL)-2, combined with antigen receptor-mediated activation, substantially rescues the suppression defects. WKO nTreg cells are also defective in the secretion of the immunomodulatory cytokine IL-10. Overall, our data reveal a critical role for WASP in nTreg cell function and implicate nTreg cell dysfunction in the autoimmunity associated with WASP deficiency. [Abstract/Link to Full Text]

Marangoni F, Trifari S, Scaramuzza S, Panaroni C, Martino S, Notarangelo LD, Baz Z, Metin A, Cattaneo F, Villa A, Aiuti A, Battaglia M, Roncarolo MG, Dupr� L
WASP regulates suppressor activity of human and murine CD4(+)CD25(+)FOXP3(+) natural regulatory T cells.
J Exp Med. 2007 Feb 19;204(2):369-80.
A large proportion of Wiskott-Aldrich syndrome (WAS) patients develop autoimmunity and allergy. CD4(+)CD25(+)FOXP3(+) natural regulatory T (nTreg) cells play a key role in peripheral tolerance to prevent immune responses to self-antigens and allergens. Therefore, we investigated the effect of WAS protein (WASP) deficiency on the distribution and suppressor function of nTreg cells. In WAS(-/-) mice, the steady-state distribution and phenotype of nTreg cells in the thymus and spleen were normal. However, WAS(-/-) nTreg cells engrafted poorly in immunized mice, indicating perturbed homeostasis. Moreover, WAS(-/-) nTreg cells failed to proliferate and to produce transforming growth factor beta upon T cell receptor (TCR)/CD28 triggering. WASP-dependent F-actin polarization to the site of TCR triggering might not be involved in WAS(-/-) nTreg cell defects because this process was also inefficient in wild-type (WT) nTreg cells. Compared with WT nTreg cells, WAS(-/-) nTreg cells showed reduced in vitro suppressor activity on both WT and WAS(-/-) effector T cells. Similarly, peripheral nTreg cells were present at normal levels in WAS patients but failed to suppress proliferation of autologous and allogeneic CD4(+) effector T cells in vitro. Thus, WASP appears to play an important role in the activation and suppressor function of nTreg cells, and a dysfunction or incorrect localization of nTreg cells may contribute to the development of autoimmunity in WAS patients. [Abstract/Link to Full Text]

Alcaide P, Jones TG, Lord GM, Glimcher LH, Hallgren J, Arinobu Y, Akashi K, Paterson AM, Gurish MA, Luscinskas FW
Dendritic cell expression of the transcription factor T-bet regulates mast cell progenitor homing to mucosal tissue.
J Exp Med. 2007 Feb 19;204(2):431-9.
The transcription factor T-bet was identified in CD4(+) T cells, and it controls interferon gamma production and T helper type 1 cell differentiation. T-bet is expressed in certain other leukocytes, and we recently showed (Lord, G.M., R.M. Rao, H. Choe, B.M. Sullivan, A.H. Lichtman, F.W. Luscinskas, and L.H. Glimcher. 2005. Blood. 106:3432-3439) that it regulates T cell trafficking. We examined whether T-bet influences homing of mast cell progenitors (MCp) to peripheral tissues. Surprisingly, we found that MCp homing to the lung or small intestine in T-bet(-/-) mice is reduced. This is reproduced in adhesion studies using bone marrow-derived MCs (BMMCs) from T-bet(-/-) mice, which showed diminished adhesion to mucosal addresin cellular adhesion molecule-1 (MAdCAM-1) and vascular cell adhesion molecule-1 (VCAM-1), endothelial ligands required for MCp intestinal homing. MCp, their precursors, and BMMCs do not express T-bet, suggesting that T-bet plays an indirect role in homing. However, adoptive transfer experiments revealed that T-bet expression by BM cells is required for MCp homing to the intestine. Furthermore, transfer of WT BM-derived dendritic cells (DCs) to T-bet(-/-) mice restores normal MCp intestinal homing in vivo and MCp adhesion to MAdCAM-1 and VCAM-1 in vitro. Nonetheless, T-bet(-/-) mice respond vigorously to intestinal infection with Trichinella spiralis, eliminating a role for T-bet in MC recruitment to sites of infection and their activation and function. Therefore, remarkably, T-bet expression by DCs indirectly controls MCp homing to mucosal tissues. [Abstract/Link to Full Text]

Jankovic D, Kullberg MC, Feng CG, Goldszmid RS, Collazo CM, Wilson M, Wynn TA, Kamanaka M, Flavell RA, Sher A
Conventional T-bet(+)Foxp3(-) Th1 cells are the major source of host-protective regulatory IL-10 during intracellular protozoan infection.
J Exp Med. 2007 Feb 19;204(2):273-83.
Although interferon gamma (IFN-gamma) secretion is essential for control of most intracellular pathogens, host survival often also depends on the expression of interleukin 10 (IL-10), a cytokine known to counteract IFN-gamma effector functions. We analyzed the source of regulatory IL-10 in mice infected with the protozoan parasite Toxoplasma gondii. Unexpectedly, IFN-gamma-secreting T-bet(+)Foxp3(-) T helper type 1 (Th1) cells were found to be the major producers of IL-10 in these animals. Further analysis revealed that the same IL-10(+)IFN-gamma(gamma) population displayed potent effector function against the parasite while, paradoxically, also inducing profound suppression of IL-12 production by antigen-presenting cells. Although at any given time point only a fraction of the cells appeared to simultaneously produce IL-10 and IFN-gamma, IL-10 production could be stimulated in IL-10(-)IFN-gamma(+) cells by further activation in vitro. In addition, experiments with T. gondii-specific IL-10(+)IFN-gamma(+) CD4 clones revealed that although IFN-gamma expression is imprinted and triggered with similar kinetics regardless of the state of Th1 cell activation, IL-10 secretion is induced more rapidly from recently activated than from resting cells. These findings indicate that IL-10 production by CD4(+) T lymphocytes need not involve a distinct regulatory Th cell subset but can be generated in Th1 cells as part of the effector response to intracellular pathogens. [Abstract/Link to Full Text]

Behfar A, Perez-Terzic C, Faustino RS, Arrell DK, Hodgson DM, Yamada S, Puceat M, Niederl�nder N, Alekseev AE, Zingman LV, Terzic A
Cardiopoietic programming of embryonic stem cells for tumor-free heart repair.
J Exp Med. 2007 Feb 19;204(2):405-20.
Embryonic stem cells have the distinct potential for tissue regeneration, including cardiac repair. Their propensity for multilineage differentiation carries, however, the liability of neoplastic growth, impeding therapeutic application. Here, the tumorigenic threat associated with embryonic stem cell transplantation was suppressed by cardiac-restricted transgenic expression of the reprogramming cytokine TNF-alpha, enhancing the cardiogenic competence of recipient heart. The in vivo aptitude of TNF-alpha to promote cardiac differentiation was recapitulated in embryoid bodies in vitro. The procardiogenic action required an intact endoderm and was mediated by secreted cardio-inductive signals. Resolved TNF-alpha-induced endoderm-derived factors, combined in a cocktail, secured guided differentiation of embryonic stem cells in monolayers produce cardiac progenitors termed cardiopoietic cells. Characterized by a down-regulation of oncogenic markers, up-regulation, and nuclear translocation of cardiac transcription factors, this predetermined population yielded functional cardiomyocyte progeny. Recruited cardiopoietic cells delivered in infarcted hearts generated cardiomyocytes that proliferated into scar tissue, integrating with host myocardium for tumor-free repair. Thus, cardiopoietic programming establishes a strategy to hone stem cell pluripotency, offering a tumor-resistant approach for regeneration. [Abstract/Link to Full Text]

Anderson CF, Oukka M, Kuchroo VJ, Sacks D
CD4(+)CD25(-)Foxp3(-) Th1 cells are the source of IL-10-mediated immune suppression in chronic cutaneous leishmaniasis.
J Exp Med. 2007 Feb 19;204(2):285-97.
Nonhealing forms of leishmaniasis in humans are commonly associated with elevated levels of the deactivating cytokine IL-10, and in the mouse, normally chronic infections can be cleared in the absence of IL-10. Using a Leishmania major strain that produces nonhealing dermal lesions in a T helper type 1 (Th1) cell-polarized setting, we have analyzed the cellular sources of IL-10 and their relative contribution to immune suppression. IL-10 was produced by innate cells, as well as CD4(+)CD25(+)Foxp3(+) and CD4(+)CD25(-)Foxp3(-) T cells in the chronic lesion. Nonetheless, only IL-10 production by antigen-specific CD4(+)CD25(-)Foxp3(-) T cells, the majority of which also produced IFN-gamma, was necessary for suppression of acquired immunity in Rag(-/-) reconstituted mice. Surprisingly, Rag(-/-) mice reconstituted with naive CD4(+) T cells depleted of natural T regulatory cells developed more severe infections, associated with elevated levels of IL-10 and, especially, Th2 cytokines in the site. The data demonstrate that IL-10-producing Th1 cells, activated early in a strong inflammatory setting as a mechanism of feedback control, are the principal mediators of T cell-derived IL-10-dependent immune suppression in a chronic intracellular infection. [Abstract/Link to Full Text]

Andersen-Nissen E, Smith KD, Bonneau R, Strong RK, Aderem A
A conserved surface on Toll-like receptor 5 recognizes bacterial flagellin.
J Exp Med. 2007 Feb 19;204(2):393-403.
The molecular basis for Toll-like receptor (TLR) recognition of microbial ligands is unknown. We demonstrate that mouse and human TLR5 discriminate between different flagellins, and we use this difference to map the flagellin recognition site on TLR5 to 228 amino acids of the extracellular domain. Through molecular modeling of the TLR5 ectodomain, we identify two conserved surface-exposed regions. Mutagenesis studies demonstrate that naturally occurring amino acid variation in TLR5 residue 268 is responsible for human and mouse discrimination between flagellin molecules. Mutations within one conserved surface identify residues D295 and D367 as important for flagellin recognition. These studies localize flagellin recognition to a conserved surface on the modeled TLR5 structure, providing detailed analysis of the interaction of a TLR with its ligand. These findings suggest that ligand binding at the beta sheets results in TLR activation and provide a new framework for understanding TLR-agonist interactions. [Abstract/Link to Full Text]

Hammad H, Kool M, Soulli� T, Narumiya S, Trottein F, Hoogsteden HC, Lambrecht BN
Activation of the D prostanoid 1 receptor suppresses asthma by modulation of lung dendritic cell function and induction of regulatory T cells.
J Exp Med. 2007 Feb 19;204(2):357-67.
Prostaglandins (PGs) can enhance or suppress inflammation by acting on different receptors expressed by hematopoietic and nonhematopoietic cells. Prostaglandin D(2) binds to the D prostanoid (DP)1 and DP2 receptor and is seen as a critical mediator of asthma causing vasodilation, bronchoconstriction, and inflammatory cell influx. Here we show that inhalation of a selective DP1 agonist suppresses the cardinal features of asthma by targeting the function of lung dendritic cells (DCs). In mice treated with DP1 agonist or receiving DP1 agonist-treated DCs, there was an increase in Foxp3(+) CD4(+) regulatory T cells that suppressed inflammation in an interleukin 10-dependent way. These effects of DP1 agonist on DCs were mediated by cyclic AMP-dependent protein kinase A. We furthermore show that activation of DP1 by an endogenous ligand inhibits airway inflammation as chimeric mice with selective hematopoietic loss of DP1 had strongly enhanced airway inflammation and antigen-pulsed DCs lacking DP1 were better at inducing airway T helper 2 responses in the lung. Triggering DP1 on DCs is an important mechanism to induce regulatory T cells and to control the extent of airway inflammation. This pathway could be exploited to design novel treatments for asthma. [Abstract/Link to Full Text]

Besseyrias V, Fiorini E, Strobl LJ, Zimber-Strobl U, Dumortier A, Koch U, Arcangeli ML, Ezine S, Macdonald HR, Radtke F
Hierarchy of Notch-Delta interactions promoting T cell lineage commitment and maturation.
J Exp Med. 2007 Feb 19;204(2):331-43.
Notch1 (N1) receptor signaling is essential and sufficient for T cell development, and recently developed in vitro culture systems point to members of the Delta family as being the physiological N1 ligands. We explored the ability of Delta1 (DL1) and DL4 to induce T cell lineage commitment and/or maturation in vitro and in vivo from bone marrow (BM) precursors conditionally gene targeted for N1 and/or N2. In vitro DL1 can trigger T cell lineage commitment via either N1 or N2. N1- or N2-mediated T cell lineage commitment can also occur in the spleen after short-term BM transplantation. However, N2-DL1-mediated signaling does not allow further T cell maturation beyond the CD25(+) stage due to a lack of T cell receptor beta expression. In contrast to DL1, DL4 induces and supports T cell commitment and maturation in vitro and in vivo exclusively via specific interaction with N1. Moreover, comparative binding studies show preferential interaction of DL4 with N1, whereas binding of DL1 to N1 is weak. Interestingly, preferential N1-DL4 binding reflects reduced dependence of this interaction on Lunatic fringe, a glycosyl transferase that generally enhances the avidity of Notch receptors for Delta ligands. Collectively, our results establish a hierarchy of Notch-Delta interactions in which N1-DL4 exhibits the greatest capacity to induce and support T cell development. [Abstract/Link to Full Text]

Dunn SE, Ousman SS, Sobel RA, Zuniga L, Baranzini SE, Youssef S, Crowell A, Loh J, Oksenberg J, Steinman L
Peroxisome proliferator-activated receptor (PPAR)alpha expression in T cells mediates gender differences in development of T cell-mediated autoimmunity.
J Exp Med. 2007 Feb 19;204(2):321-30.
Peroxisome proliferator-activated receptor (PPAR)alpha is a nuclear receptor that mediates gender differences in lipid metabolism. PPARalpha also functions to control inflammatory responses by repressing the activity of nuclear factor kappaB (NF-kappaB) and c-jun in immune cells. Because PPARalpha is situated at the crossroads of gender and immune regulation, we hypothesized that this gene may mediate sex differences in the development of T cell-mediated autoimmune disease. We show that PPARalpha is more abundant in male as compared with female CD4(+) cells and that its expression is sensitive to androgen levels. Genetic ablation of this gene selectively removed the brake on NF-kappaB and c-jun activity in male T lymphocytes, resulting in higher production of interferon gamma and tumor necrosis factor (but not interleukin 17), and lower production of T helper (Th)2 cytokines. Upon induction of experimental autoimmune encephalomyelitis, male but not female PPARalpha(-/-) mice developed more severe clinical signs that were restricted to the acute phase of disease. These results suggest that males are less prone to develop Th1-mediated autoimmunity because they have higher T cell expression of PPARalpha. [Abstract/Link to Full Text]

Boissonnas A, Fetler L, Zeelenberg IS, Hugues S, Amigorena S
In vivo imaging of cytotoxic T cell infiltration and elimination of a solid tumor.
J Exp Med. 2007 Feb 19;204(2):345-56.
Although the immune system evolved to fight infections, it may also attack and destroy solid tumors. In most cases, tumor rejection is initiated by CD8(+) cytotoxic T lymphocytes (CTLs), which infiltrate solid tumors, recognize tumor antigens, and kill tumor cells. We use a combination of two-photon intravital microscopy and immunofluorescence on ordered sequential sections to analyze the infiltration and destruction of solid tumors by CTLs. We show that in the periphery of a thymoma growing subcutaneously, activated CTLs migrate with high instantaneous velocities. The CTLs arrest in close contact to tumor cells expressing their cognate antigen. In regions where most tumor cells are dead, CTLs resume migration, sometimes following collagen fibers or blood vessels. CTLs migrating along blood vessels preferentially adopt an elongated morphology. CTLs also infiltrate tumors in depth, but only when the tumor cells express the cognate CTL antigen. In tumors that do not express the cognate antigen, CTL infiltration is restricted to peripheral regions, and lymphocytes neither stop moving nor kill tumor cells. Antigen expression by tumor cells therefore determines both CTL motility within the tumor and profound tumor infiltration. [Abstract/Link to Full Text]

Ford ML, Koehn BH, Wagener ME, Jiang W, Gangappa S, Pearson TC, Larsen CP
Antigen-specific precursor frequency impacts T cell proliferation, differentiation, and requirement for costimulation.
J Exp Med. 2007 Feb 19;204(2):299-309.
After a brief period of antigenic stimulation, T cells become committed to a program of autonomous expansion and differentiation. We investigated the role of antigen-specific T cell precursor frequency as a possible cell-extrinsic factor impacting T cell programming in a model of allogeneic tissue transplantation. Using an adoptive transfer system to incrementally raise the precursor frequency of antigen-specific CD8(+) T cells, we found that donor-reactive T cells primed at low frequency exhibited increased cellular division, decreased development of multifunctional effector activity, and an increased requirement for CD28- and CD154-mediated costimulation relative to those primed at high frequency. The results demonstrated that recipients with low CD4(+) and CD8(+) donor-reactive T cell frequencies exhibited long-term skin graft survival upon CD28/CD154 blockade, whereas simultaneously raising the frequency of CD4(+) T cells to approximately 0.5% and CD8(+) T cells to approximately 5% precipitated graft rejection despite CD28/CD154 blockade. Antigenic rechallenge of equal numbers of cells stimulated at high or low frequency revealed that cells retained an imprint of the frequency at which they were primed. These results demonstrate a critical role for initial precursor frequency in determining the CD8(+) T cell requirement for CD28- and CD154-mediated costimulatory signals during graft rejection. [Abstract/Link to Full Text]

Allakhverdi Z, Comeau MR, Jessup HK, Yoon BR, Brewer A, Chartier S, Paquette N, Ziegler SF, Sarfati M, Delespesse G
Thymic stromal lymphopoietin is released by human epithelial cells in response to microbes, trauma, or inflammation and potently activates mast cells.
J Exp Med. 2007 Feb 19;204(2):253-8.
Compelling evidence suggests that the epithelial cell-derived cytokine thymic stromal lymphopoietin (TSLP) may initiate asthma or atopic dermatitis through a dendritic cell-mediated T helper (Th)2 response. Here, we describe how TSLP might initiate and aggravate allergic inflammation in the absence of T lymphocytes and immunoglobulin E antibodies via the innate immune system. We show that TSLP, synergistically with interleukin 1 and tumor necrosis factor, stimulates the production of high levels of Th2 cytokines by human mast cells (MCs). We next report that TSLP is released by primary epithelial cells in response to certain microbial products, physical injury, or inflammatory cytokines. Direct epithelial cell-mediated, TSLP-dependent activation of MCs may play a central role in "intrinsic" forms of atopic diseases and explain the aggravating role of infection and scratching in these diseases. [Abstract/Link to Full Text]

Svensson CI, Zattoni M, Serhan CN
Lipoxins and aspirin-triggered lipoxin inhibit inflammatory pain processing.
J Exp Med. 2007 Feb 19;204(2):245-52.
Inflammatory conditions can lead to debilitating and persistent pain. This hyperalgesia reflects sensitization of peripheral terminals and facilitation of pain signaling at the spinal level. Studies of peripheral systems show that tissue injury triggers not only inflammation but also a well-orchestrated series of events that leads to reversal of the inflammatory state. In this regard, lipoxins represent a unique class of lipid mediators that promote resolution of inflammation. The antiinflammatory role of peripheral lipoxins raises the hypothesis that similar neuraxial systems may also down-regulate injury-induced spinal facilitation of pain processing. We report that the lipoxin A(4) receptor is expressed on spinal astrocytes both in vivo and in vitro and that spinal delivery of lipoxin A(4), as well as stable analogues, attenuates inflammation-induced pain. Furthermore, activation of extracellular signal-regulated kinase and c-Jun N-terminal kinase in astrocytes, which has been indicated to play an important role in spinal pain processing, was attenuated in the presence of lipoxins. This linkage opens the possibility that lipoxins regulate spinal nociceptive processing though their actions upon astrocytic activation. Targeting mechanisms that counterregulate the spinal consequences of persistent peripheral inflammation provide a novel endogenous mechanism by which chronic pain may be controlled. [Abstract/Link to Full Text]

Gourzi P, Leonova T, Papavasiliou FN
Viral induction of AID is independent of the interferon and the Toll-like receptor signaling pathways but requires NF-kappaB.
J Exp Med. 2007 Feb 19;204(2):259-65.
Activation-induced cytidine deaminase (AID) is expressed in germinal centers of lymphoid organs during immunoglobulin diversification, in bone marrow B cells after infection with Abelson murine leukemia retrovirus (Ab-MLV), and in human B cells after infection by hepatitis C virus. To understand how viruses signal AID induction in the host we asked whether the AID response was abrogated in cells deficient in the interferon pathway or in signaling via the Toll-like receptors. Here we show that AID is not an interferon responsive gene and abrogation of Toll-like receptor signaling does not diminish the AID response. However, we found that NF-kappaB was required for expression of virally induced AID. Since NF-kappaB binds and activates the AID promoter, these results mechanistically link viral infection with AID transcription. Thus, induction of AID by viruses could be the result of several signaling pathways that culminate in NF-kappaB activation, underscoring the versatility of this host defense program. [Abstract/Link to Full Text]

Gross S, Tilly P, Hentsch D, Vonesch JL, Fabre JE
Vascular wall-produced prostaglandin E2 exacerbates arterial thrombosis and atherothrombosis through platelet EP3 receptors.
J Exp Med. 2007 Feb 19;204(2):311-20.
Prostanoids, bioactive lipids derived from arachidonic acid (AA), are important for vascular homeostasis. Among them, prostaglandin E2 (PGE2) enhances aggregation of platelets submaximally stimulated in vitro. This results from activation of EP3, one of the four PGE2 receptors, which decreases the threshold at which agonists activate platelets to aggregate. Although PGE2 altered venous thrombosis induced by administration of AA, its role in pathophysiopathological conditions has remained speculative. We report that arterial walls subjected to inflammatory stimuli produce PGE2. In several models, we show that PGE2 produced by the arterial wall facilitates arterial thrombosis. Next, we detected PGE2 in mouse atherosclerotic plaques. We demonstrate that this plaque-produced PGE2 is not altered and is still able to activate EP3. In addition, we present evidence that PGE2 can leave the plaque and activate EP3 on blood platelets. Consistent with these findings, we observed that atherothrombosis induced in vivo by mechanical rupture of the plaque was drastically decreased when platelets lacked EP3. In conclusion, PGE2 facilitates the initiation of arterial thrombosis and, hence, contributes to atherothrombosis. Inhibition of the platelet EP3 receptor should improve prevention of atherothrombosis. [Abstract/Link to Full Text]

Bashyam H
IFNgamma: issuing macrophages a license to kill.
J Exp Med. 2007 Jan 22;204(1):3.
T cells tell macrophages when to start making the toxic soup of lysosomal enzymes, reactive oxygen species, and nitric oxide that destroys intracellular pathogens. In 1983, Carl Nathan proved that this start signal comes in the form of the secreted cytokine IFNgamma. [Abstract/Link to Full Text]