Rachel L. Ruhlen

Estradiol and Bisphenol A Stimulate Androgen Receptor and Estrogen Receptor Gene Expression in Fetal Mouse Prostate Mesenchyme Cells

Background Hormonal alterations during development have lifelong effects on the prostate gland. Endogenous estrogens, including 17β-estradiol (E2), and synthetic estrogenic endocrine disruptors, such as bisphenol A (BPA), have similar effects on prostate development. Increasing exposure to estrogens within the low-dose, physiologic range results in permanent increases in the size and androgen responsiveness of the prostate, whereas exposure within the high-dose, pharmacologic range has the opposite effects. Objectives We tested the hypothesis that the low-dose effects of estrogens on the developing prostate are associated with increased expression of androgen receptor (Ar) and estrogen receptor 1 (α) (Esr1) genes in mesenchyme cells. Methods Ar and Esr1 mRNA levels were quantified in primary cultures of fetal mouse prostate mesenchyme cells treated with E2 and BPA. Discussion Ar and Esr1 mRNA expression increased in response to E2, with thresholds of 0.001 and 0.037 nM, respectively; and in response to BPA, with a threshold of 1 nM for both mRNAs. We did not observe the expected inhibition of Ar mRNA expression by pharmacologic levels of E2 relative to unexposed cells. Conclusions The observed induction of gene expression occurred at concentrations within the range of free E2 previously shown to permanently increase prostate size, thus supporting the involvement of direct effects of estrogens on gene expression in prostate mesenchyme. The effects of BPA occurred within the range of concentrations currently measured in human serum, demonstrating the vulnerability of developing tissues to xenoestrogens.

Soy Isoflavones Have an Antiestrogenic Effect and Alter Mammary Promoter Hypermethylation in Healthy Premenopausal Women

We determined if soy isoflavones have dose-related estrogenic and methylation effects. Thirty-four healthy premenopausal women were randomized to 40 mg or 140 mg isoflavones daily through one menstrual cycle. Breast specific and systemic estrogenic effects were assessed measuring the estrogenic marker complement (C)3 and changes in cytology, whereas methylation assessment of 5 cancer related genes (p16, RASSF1A, RAR β 2, ER, and CCND2) was performed on intraductal specimens. Serum genistein significantly increased after consuming both isoflavone doses. Cytology did not significantly change at either isoflavone dose. Serum C3 levels posttreatment were inversely related to change in serum genistein ( r =–0.76, P = 0.0045) in women consuming low but not high dose isoflavones. The RAR β 2 hypermethylation increase posttreatment correlated with the posttreatment genistein level considering the entire group ( r = 0.67, P = 0.0017) and those receiving high-dose isoflavones ( r = 0.68, P = 0.021). At the low but not the high isoflavone dose, CCND2 hypermethylation increase correlated with posttreatment genistein levels ( r = 0.79, P = 0.011). In summary, the inverse correlation between C3 and genistein suggests an antiestrogenic effect. Isoflavones induced dose-specific changes in RAR β 2 and CCND2 gene methylation, which correlated with genistein levels. This work provides novel insights into estrogenic and methylation effects of dietary isoflavones.

Low Phytoestrogen Levels in Feed Increase Fetal Serum Estradiol Resulting in the “Fetal Estrogenization Syndrome” and Obesity in CD-1 Mice

Background Although estrogenic chemicals can disrupt development of the reproductive system, there is debate about whether phytoestrogens in soy are beneficial, benign, or harmful. Objectives We compared reproductive and metabolic characteristics in male and female mice reared and maintained on non-soy low-phytoestrogen feed or soy-based high-phytoestrogen feed. Methods The low-phytoestrogen diet was non-soy PMI 5K96 (verified casein diet), and the high-phytoestrogen diet consisted of soy-based PMI 5008 during pregnancy and lactation and soy-based PMI 5001 maintenance feed after weaning. Results In fetuses whose mothers consumed the low-phytoestrogen PMI 5K96 feed, we found a paradoxical significant elevation in endogenous serum estradiol, which was associated postnatally with adverse reproductive outcomes referred to as the “fetal estrogenization syndrome (FES)”. In females, this syndrome included early puberty and increased uterine responsiveness to estrogen, and in males, it included reduced testis, epididymis, and seminal vesicle size, but an enlarged prostate. The low-phytoestrogen–fed males and females were lighter at birth, but, between weaning and adulthood, they became obese and developed abnormally high serum leptin levels; these males, but not females, showed impaired glucose regulation. Conclusions Removing phytoestrogens from mouse feed produces an obese phenotype consistent with metabolic syndrome, and the associated reproductive system abnormalities are consistent with FES due to elevated endogenous fetal estradiol. Laboratory rodents may have become adapted to high-phytoestrogen intake over many generations of being fed soy-based commercial feed; removing all phytoestrogens from feed leads to alterations that could disrupt many types of biomedical research.

Estrogenic Environmental Chemicals and Drugs: Mechanisms for Effects on the Developing Male Urogenital System

Development and differentiation of the prostate from the fetal urogenital sinus (UGS) is dependent on androgen action via androgen receptors (AR) in the UGS mesenchyme. Estrogens are not required for prostate differentiation but do act to modulate androgen action. In mice exposure to exogenous estrogen during development results in permanent effects on adult prostate size and function, which is mediated through mesenchymal estrogen receptor (ER) alpha. For many years estrogens were thought to inhibit prostate growth because estrogenic drugs studied were administered at very high concentrations that interfered with normal prostate development. There is now extensive evidence that exposure to estrogen at very low concentrations during the early stages of prostate differentiation can stimulate fetal/neonatal prostate growth and lead to prostate disease in adulthood. Bisphenol A (BPA) is an environmental endocrine disrupting chemical that binds to both ER receptor subtypes as well as to AR. Interest in BPA has increased because of its prevalence in the environment and its detection in over 90% of people in the USA. In tissue culture of fetal mouse UGS mesenchymal cells, BPA and estradiol stimulated changes in the expression of several genes. We discuss here the potential involvement of estrogen in regulating signaling pathways affecting cellular functions relevant to steroid hormone signaling and metabolism and to inter- and intra-cellular communications that promote cell growth. The findings presented here provide additional evidence that BPA and the estrogenic drug ethinylestradiol disrupt prostate development in male mice at administered doses relevant to human exposures.

Black Cohosh Does Not Exert an Estrogenic Effect on the Breast

Abstract Womens Health Initiative findings indicate that hormone replacement therapy may increase breast cancer and cardiovascular disease risk. Black cohosh extract (BCE) is a popular alternative that reduced menopausal symptoms in several clinical trials. Preclinical studies have addressed the estrogenic properties of BCE, with conflicting results. The estrogenic influence of BCE on the breast has not been investigated. Black cohosh is standardized to triterpenes, but the activity and mechanism of action of these compounds are unknown. The study goals were to determine 1) triterpene content of 2 commercially available BCE preparations and 2) the effect of BCE on circulating and breast-specific estrogenic markers. Two black cohosh preparations were analyzed for triterpene content. Postmenopausal women took BCE for 12 wk followed by a 12-wk washout. One BCE preparation contained trace amounts and another contained 2.5% triterpenes. Women taking BCE with 2.5% triterpenes experienced relief of menopausal symptoms, with reversion toward baseline after washout. BCE had no effect on estrogenic markers in serum and no effect on pS2 or cellular morphology in nipple aspirate fluid. Triterpene content in commercially available black cohosh preparations varies. BCE standardized to 2.5% triterpenes relieved menopausal symptoms without systemic or breast-specific estrogenic effects.

Biomarkers associated with breast cancer are associated with obesity.

BACKGROUND Obesity is linked to the development of postmenopausal breast cancer, and some studies indicate obesity predicts a worse prognosis for premenopausal women who develop the disease. It was our hypothesis that proteins associated with breast cancer would be associated with body mass index (BMI). METHODS We searched our database of women enrolled in breast health translational research trials for information on BMI and markers predictive of breast cancer (basic fibroblast growth factor (bFGF), prostate-specific antigen (PSA), human kallikrein (hK)2, and urinary plasminogen activator (uPA). Information on BMI and one or more nipple aspirate fluid (NAF) or serum biomarkers was available from 382 women. RESULTS In this data set, NAF and serum levels of PSA (nPSA and sPSA), and NAF levels hK2, bFGF and uPA were each associated with pre- and/or postmenopausal breast cancer. sPSA was inversely associated with BMI in both pre- (r=-.56, p=.001) and postmenopausal women (r=-.62, p=.0035) without breast cancer. This association was lost when controlling for plasma volume. In women without breast cancer, NAF bFGF (p=.07, premenopausal subjects) and NAF hK2 (p=.09, postmenopausal subjects) were borderline associated with BMI. In women with breast cancer, nPSA was inversely (r=-.53, p=.049) associated with BMI in premenopausal women and directly associated with BMI in postmenopausal women (r=.37, p=.017). nPSA trended higher in hormone sensitive cancers, especially those that expressed progesterone receptor (p=.059). CONCLUSIONS sPSA was inversely associated with BMI in all pre- and postmenopausal women and specifically in pre- and postmenopausal women without breast cancer. NAF PSA was associated with BMI in pre- and postmenopausal women with breast cancer. Evaluating the change in PSA with changes in weight may provide clues regarding a subjects breast cancer risk.

Differential expression of cancer associated proteins in breast milk based on age at first full term pregnancy.

BackgroundFirst full term pregnancy (FFTP) completed at a young age has been linked to low long term breast cancer risk, whereas late FFTP pregnancy age confers high long term risk, compared to nulliparity. Our hypothesis was that proteins linked to breast cancer would be differentially expressed in human milk collected at three time points during lactation based on age at FFTP.MethodsWe analyzed breast milk from 72 lactating women. Samples were collected within 10 days of the onset of lactation (baseline-BL), two months after lactation started and during breast weaning (W). We measured 16 proteins (11 kallikreins (KLKs), basic fibroblast growth factor, YKL-40, neutrophil gelatinase-associated lipocalin and transforming growth factor (TGF) β-1 and -2) associated with breast cancer, most known to be secreted into milk.ResultsDuring lactation there was a significant change in the expression of 14 proteins in women < 26 years old and 9 proteins in women > = 26 at FFTP. The most significant (p < .001) changes from BL to W in women divided by FFTP age (< 26 vs. > = 26) were in KLK3,6, 8, and TGFβ2 in women < 26; and KLK6, 8, and TGFβ2 in women > = 26. There was a significant increase (p = .022) in KLK8 expression from BL to W depending on FFTP age. Examination of DNA methylation in the promoter region of KLK6 revealed high levels of methylation that did not explain the observed changes in protein levels. On the other hand, KLK6 and TGFβ1 expression were significantly associated (r2 = .43, p = .0050).ConclusionsThe expression profile of milk proteins linked to breast cancer is influenced by age at FFTP. These proteins may play a role in future cancer risk.

Choice of animal feed can alter fetal steroid levels and mask developmental effects of endocrine disrupting chemicals

Exposure of fetuses to endocrine disrupting chemicals (EDCs), such as the estrogenic drug diethylstilbestrol (DES), disrupts development of the reproductive system and affects other aspects of adult phenotype including diseases, consistent with the developmental origins of health and disease hypothesis. To determine whether diet could influence the effects of DES, we compared mice fed a commonly used combination of soy-based Purina 5008 (breeding and lactation) and 5001 (post-weaning) with mice fed soy-based Purina 5002 throughout life. We exposed fetal CD-1 mice (F1) in utero on different feeds to a 0 (controls), low (0.1 mg/kg/day) or high (50 mg/kg/day) dose of DES via feeding the dam (F0) on gestation days 11–17. Compared to 5008, 5002 feed significantly increased serum estradiol in control fetuses. On 5008 (but not 5002) feed, DES significantly increased fetal serum estradiol at a low dose and reduced it at a high dose. Diet influenced the effects of in utero DES on F1 female onset of puberty and the uterine response to estradiol (an inverted-U dose–response relationship seen for DES on uterine weight with 5008/5001 feed was not observed with 5002). Both low- and high-dose DES reduced daily sperm production (DSP) in adult F1 males on 5008/5001 feed, whereas males fed 5002 showed no DES-induced reduction in DSP. Thus, we observed a number of low-dose effects of in utero DES exposure on Purina 5008/5001 feed that were not observed using Purina 5002, a feed commonly used in industry-funded toxicological studies conducted for regulatory purposes. Received 24 August 2010; Revised 26 October 2010; Accepted 2 December 2010; First published online 28 January 2011

Celecoxib concentration predicts decrease in prostaglandin E2 concentrations in nipple aspirate fluid from high risk women

BackgroundEpidemiologic studies suggest that long term low dose celecoxib use significantly lowers breast cancer risk. We previously demonstrated that 400 mg celecoxib taken twice daily for 2 weeks lowered circulating plasma and breast nipple aspirate fluid (NAF) prostaglandin (PG)E2 concentrations in post- but not premenopausal high risk women. We hypothesized that circulating concentrations of celecoxib influenced PGE2 response, and that plasma levels of the drug are influenced by menopausal status. To address these hypotheses, the aims of the study were to determine: 1) if circulating plasma concentrations of celecoxib correlated with the change in plasma or NAF PGE2 concentrations from baseline to end of treatment, and 2) whether menopausal status influenced circulating levels of celecoxib.MethodsMatched NAF and plasma were collected from 46 high risk women who were administered celecoxib twice daily for two weeks, 20 subjects receiving 200 mg and 26 subjects 400 mg of the agent. NAF and plasma samples were collected before and 2 weeks after taking celecoxib.ResultsIn women taking 400 mg bid celecoxib, plasma concentrations of the agent correlated inversely with the change in NAF PGE2 levels from pre- to posttreatment. Nonsignificant trends toward higher celecoxib levels were observed in post- compared to premenopausal women. There was a significant decrease in NAF but not plasma PGE2 concentrations in postmenopausal women who took 400 mg celecoxib (p = 0.03).ConclusionIn high risk women taking 400 mg celecoxib twice daily, plasma concentrations of celecoxib correlated with downregulation of PGE2 production by breast tissue. Strategies synergistic with celecoxib to downregulate PGE2 are of interest, in order to minimize the celecoxib dose required to have an effect.

Black cohosh: Insights into its mechanism(s) of action

Objective: To investigate the effects of a daily multinutrient supplement on plasma indicators of glycemic and lipemic control and psychological wellbeing in type 2 diabetic patients. Design: Double-blind, randomised, cross-over pilot intervention study. Subjects: Twenty-nine subjects (15 males and 14 females) with non-insulin-dependent type 2 diabetes. Intervention: Either a multinutrient supplement or placebo were provided daily during two intervention periods of 3 months separated by a 4 week washout. Results: There were no significant changes of multinutrient treatment compared with placebo in HbA1c, fasting or postprandial plasma glucose and insulin concentrations and fasting plasma lipid concentrations. Using a validated wellbeing questionnaire (W-BQ 22) designed for diabetic subjects, the multinutrient supplement resulted in improvements in the secondary outcome of wellbeing of the volunteers in terms of anxiety (p = 0.020), vitality (p = 0.013) and general wellbeing (p = 0.021), relative to placebo. Conclusions: Findings from this pilot study suggest that a multinutrient supplement may enhance the wellbeing of diabetic patients, even in the absence of a significant improvement in clinical parameters. If substantiated in a full clinical study the results would have important implications for the prevention of late complications of diabetes, as psychological factors can hinder successful management of the condition and adversely affect metabolic control.From the aerial parts of Ruta chalepensis L., grown in Jordan, two furanocoumarins (bergapten and chalepensin), one flavonoid glycoside (rutin) as well as several minor compounds have been isolated. The structural elucidation of these compounds was established based on spectral data (UV, IR, MS,1H-NMR and 13C-NMR). In Jordan, R. chalepensis is recommended for the treatment of rheumatism, mental disorders and menstrual problems. Fresh and dried leaves are used as flavoring agent in food and beverages. Antiplatelet activities of the crude methanolic and ethylacetate extracts in addition to the three isolated major compounds were measured by the aggrometric method according to Beretz and Casenave. Optical aggregometer connected to dual channel recorder was used for measuring aggregation. Both, ethylacetate and methanol extracts inhibited ADP- induced platelet aggregation (ADP-IA) of human blood. However, only ethylacetate extract was able to induce 50% inhibition of collagen-induced platelet aggregation (Co-IA) platelet rich plasma. Bergapten was more active against ADP-IA compared to chalepensin while the latter was more active against Co-IA compared to bergapten.Background: Hypertension is a major health problem with serious medical and financial consequences. Experimental studies in animals and clinical studies in humans have demonstrated that acupuncture can reduce blood pressure significantly in hypertensive patients. The objective was to assess the effect of acupuncture on blood pressure in hypertensive patients treated at a complementary medicine clinic. Methods: Blood pressure values measured before and following acupuncture were recorded from the charts of hypertensive patients who came to the clinic for treatment of other problems. The therapy used was the Kiiko Matsumoto technique for blood pressure imbalance. Results: Twenty-nine patients were studied (18 [62%] women). The mean age was 58.5 ± 16.3 years. Systolic blood pressure dropped significantly as a result of the treatment and there was a non-significant trend to reduced diastolic pressure. Weekly acupuncture therapy led to a continuous reduction in systolic blood pressure. Conclusions: Acupuncture has a beneficial effect on hypertension, particularly on systolic pressure. Further studies with larger study groups for longer periods of time can confirm this observation and contribute to our understanding of combination therapy with acupuncture and conventional medications for hypertension.Background: Population-based data about utilization of complementary and alternative medicine (CAM) among those with chronic conditions is lacking. Objective: To describe whether CAM use by California adults with cancer and other chronic conditions reflects condition specific patterns or a general tendency to use CAM modalities. Methods: Interviews of 9,187 respondents including all participants with cancer from a prior representative survey of California households, and a stratified sample of all other respondents. Almost 74% of the respondents reported at least one chronic health problem. Results: Use of all forms of CAM among those with chronic health problems is high. Those with a diagnosis of cancer are more likely to use prayer, dietary supplements, and support groups, and less likely to use CAM providers and special diets. Overall, individuals diagnosed with most chronic problems use a similar set of CAM modalities. Demographic correlates of CAM use differ in their impact and vary according to what type of CAM is being used. Conclusions: Clinicians should be aware that while a diagnosis of cancer is associated with a greater use of some forms of CAM, overall patterns of CAM use are similar to those with most other chronic problems.We propose the formation of an International PsychoSocial and Cultural Bioinformatics Project (IPCBP) to explore the research foundations of Integrative Medical Insights (IMI) on all levels from the molecular-genomic to the psychological, cultural, social, and spiritual. Just as The Human Genome Project identified the molecular foundations of modern medicine with the new technology of sequencing DNA during the past decade, the IPCBP would extend and integrate this neuroscience knowledge base with the technology of gene expression via DNA/proteomic microarray research and brain imaging in development, stress, healing, rehabilitation, and the psychotherapeutic facilitation of existentional wellness. We anticipate that the IPCBP will require a unique international collaboration of, academic institutions, researchers, and clinical practioners for the creation of a new neuroscience of mind-body communication, brain plasticity, memory, learning, and creative processing during optimal experiential states of art, beauty, and truth. We illustrate this emerging integration of bioinformatics with medicine with a videotape of the classical 4-stage creative process in a neuroscience approach to psychotherapy.In this essay, we posit that modern medicine, in its Asclepian focus, has subordinated the need and importance of Hygieian healing and caring, and in so doing has lost a quality that is essential to medicine, and fundamental to its lasting moral value. We argue that an integrative medicine must be based upon a core philosophical foundation that re-enjoins Asclepian and Hygieian approaches in true conceptual and practical dialectic, such that integration represents a synthesis of these orientations in epistemic, humanitarian and ethical domains. While we assert that a core philosophy is critical to the development and sustainability of an integrative medicine, such claims remain vacant in the absence of some meaningful attempt to put these concepts into action. We believe that to apply such philosophical foundations, an approach is necessary that simultaneously engages education, research, practice and policy. This involves not simply studying and co-opting new (or older, more ancient) modalities in a curative paradigm, but represents a paradigm shift that requires and is based upon understanding of, and skills for the application(s) of the most appropriate types of treatment(s) to affect disease, illness and health in a patient-centered model of care.Roughly 90 years of research demonstrate the relevance of dietary nutrients for mental health. Some of the earliest research studies on nutrients relevant to mental illness observed irritability and mood problems in people known to be deficient in the B vitamins1, as well as reported positive improvements in mental illness when treated with such nutrients as manganese2,3 and nicotinic acid;4 regardless of whether or not the patients could be found to be deficient. Although interest in such studies have declined since the introduction of psychiatric medications in the 1950’s, recent work on folic acid (vitamin B9) suggests that low levels may be associated with depressive symptomatology and poor response to antidepressant medication.5 Increasing evidence about the effects of trace elements on brain and behavioral functioning is appearing as well. Zinc, copper, and magnesium may play an important modulatory role in controlling a subtype of glutamate receptor (NMDA receptor),6 glutamate being the primary transmitter for most excitatory neurons in the cerebral cortex. This NMDA receptor has been implicated in various forms of cortical functioning;7 therefore it appears that decreased levels of these nutrients may produce abnormal NMDA activity and subsequent abnormal behavior. Given the accumulating evidence from PET and fMRI imaging studies showing that schizophrenia and affective disorders are associated with abnormal cortical activity,8 it is logical to state that such conditions could result, at least in part, from abnormalities in the nutritional status of neurons. Other studies regarding the relevance of nutrients and schizophrenia have been conducted as well. Comparison studies have shown that 26 medication-free schizophrenics were found to have significantly low serum iron,9 in addition to a study in Israel where both the cerebrospinal fluid and serum of people with schizophrenia were tested to be low in magnesium.10 Still others have studied essential fatty acid-related membrane processes. Among 38 schizophrenics and 22 controls, in the cutaneous flushing response to aqueous methyl nicotinate,: 83% of the people with schizophrenia (but only 23% of the controls) exhibited the absence of a flushing response, indicative of deficient levels of arachidonic acid.11 This particular study is relevant due to the fact that some minerals (e.g. Zinc) are thought to be rate-limiting factors in essential fatty acid conversion pathways.12 High dose vitamin therapy has been studied with a number of genetic diseases. The molecular basis of disease arising from as many as one-third of the mutations in a gene is an increased Michaelis constant, or KM (decreasing binding affinity) of an enzyme for the vitamin-derived coenzyme or substrate, which in turn lowers the rate of the reaction.13 The KM is defined as the concentration of ligand required to fill one-half of the ligand binding sites. Therapeutic vitamin regimens are thought to increase intracellular (cofactor) concentration, thus activating a defective enzyme, which alleviates the primary defect and remediates the disease. The proportion of mutations in a disease gene that is responsive to high concentrations of a vitamin or substrate may be one-third or greater.14,15,16 The battle to reduce the stigma associated with nutritional therapies is still very present today, fifty years later. More commonly labeled “alternative medicine/therapy,” nutritional therapies are considered just that: an alternative, a last resort, or are not considered at all. Over the years, major medical textbooks have claimed that “routine prescription of vitamin preparations is indefensible, it is poor medical practice,17 and that “multivitamins are not necessary.18 Goodwin went so far as to say that a bias exists in this particular area; where “positive results are viewed with suspicion,” and “negative results are published in the best journals.”19 One study found that most doctors do not feel comfortable discussing alternative therapies with their patients, despite the fact that 55% of patients have requested more information about herbal (or natural) medicine.20 However, despite the presence of skeptics, criticisms and lack of information to the public, natural therapies continue to be used. We will review a number of substances and their potential use in psychiatry. Essential fatty acids Essential Fatty Acids (EFA) must be obtained either from diet or through supplementation. Premenstrual Dysphoric Disorder (PMDD) affects many women in varying degrees, ranging from only physical symptoms before the menses to varying degrees of irritability, anger, and depression. Ranging from mild to severe, PMDD can be treated with Evening Primrose Oil (EPO). EPO contains two essential precursors for prostaglandin synthesis, 70% cis-linoleic acid, and 8 to 14% gamma-linolenic acid. By providing the body with these essential fatty acids, EPO facilitates the synthesis of Prostaglandin E1 (PGE1); a substance that women with PMDD may lack in the central nervous system as well as in other tissue including the breast tissue.21 In an open trial, 18 women with PMDD of more than 1 year’s duration received 8 capsules/day of evening primrose oil in the last half of the menstrual cycles for 5 cycles.22 Irritability (p < 0.001), depression (p < 0.001), anxiety (p < 0.01), and fatigue (p < 0.01) were significantly less compared to baseline after the first cycle of treatment. Total PMS scores were significantly improved (p < 0.001). A Cochran Database review concludes that limited evidence gives support to a hypothesis suggesting that the symptoms of schizophrenia may result from altered neuronal membrane structure and metabolism.23 The latter are dependent on blood plasma levels of certain essential fatty acids (EFAs) and their metabolites. They found several studies showing that those with schizophrenia often have low levels of the particular EFAs necessary for normal nerve cell membrane metabolism. Four relatively small trials (total n = 204) showed low levels of loss to follow up and adverse effects for those taking essential fatty acids. Early results from a few trials suggest a positive effect of eicosapentaenoic acid (EPA) over placebo for scale-derived mental state outcomes. The data, however, was limited, making these results difficult to analyze and interpret with confidence. A single small study (n = 30) investigated the value of using EPA as sole treatment for people hospitalized for relapse. Results suggested that EPA could help one third of people avoid instigation of standard antipsychotic drugs for 12 weeks (RR 0.6, CI 0.4–0.91). There were no clear effects of primrose oil (omega-6) EFA supplementation. Omega-3-Fatty acids have been used as treatment for depression,24 especially eicosapentaenoic acid.25,26,27Introduction The history of the European Union goes back to the 1950’s, when a new approach in economic cooperation started between some states of the continent. As it gradually developed, the four basic freedoms (free movement of persons, goods, services, and capital) assured by the participating countries became the basis of the cooperation. The creation of the European Union in 1992 brought forth a wider sphere of authority than already existed regarding the common economic issues. Some of the provisions related to health care even gained a great publicity, like the introduction of the E111 card or the European Court decision concerning the Simap and Valencia case (as a result of the latter, the member states are bound to regard the time spent on call by doctors in primary health care teams in its entirety as working time)(1). Although matters connected to alternative medicine generally fall within the authority of the member states, policies of the EU do have a certain impact in this area as well. Since a specifi c legal regulation is absent or incomplete in several countries, an indirect unifying impact of EU activities might emerge and reach a signifi cant level (2)(3). Alternative medical practices and products are widely used and applied by the citizens of the member states (4). Due to this increasing acceptance some might experience a growing EU activity, like speeches in the parliament, consultations with EU institutions, and advisory panels (2). However promising these manifestations might be at a fi rst glance, all but the directives are not binding for the member states. Up to now, beyond the favoring voices no specifi c law or signifi cant fi nancial support appeared. Two programs, starting in 2007 might become a turning point in the relation of the EU to alternative medicine. First, the ‘‘Programme of Community Action in the Field of Health 2007–2013” attempts to support mobility between patients and providers of the member states and stresses the assurance of patients’ free choice within health care (5). Second, the Seventh Framework Programme (FP7)—that will be approved before 2006—enables the fi nancial support of research in the case of alternative medicine also (6). The legal activity of the EU by means of its provisions or specifi c policies infl uences even unintentionally, or indirectly the developmental pathways of alternative medicine. Attempting to highlight some of them, the paper discusses the impact of education, research and migration policies on alternative medicine.It has been well reported that complementary medicines can significantly alter the way the body handles conventional drugs, leading to potential fatal herb-drug interactions. The aim of the present study was to investigate the molecular mechanism of drug interactions involving St John’s wort (SJW) (Hypericum perforatum L), a popular herbal medicine widely used for depression, particularly examining changes in the expression of cytochrome P450 CYP3A, the most abundant drug metabolising CYP enzymes in man. Eighteen Sprague-Dawley (SD) rats were assigned randomly into 3 groups (n = 6/group): control, low dose and high dose (500 and 1000 mg/kg/day of SJW, equal to 1500 and 3000 μg/kg/day of Hypericin). Each group was treated with SJW or control preparation, by gastric gavage, for 14 consecutive days. Liver and intestinal CYP3A activity and protein and mRNA levels, from fi ve segments of the intestine, were examined using CYP3A-dependent erythromycin N-demethylation activity assay, quantitative immuno-blotting and real-time RT-PCR. Increase in CYP3A activity and protein level by SJW was observed in some intestinal regions, with a 3.0 fold increase in liver CYP3A activity and a 10.6 fold increase in liver CYP3A1 mRNA (p 0.05) in a dose dependent manner. The results suggested that up regulation of liver CYP3A mRNA and differential induction of intestinal CYP3A play an important role in the molecular mechanism of herb-drug interactions.The Women’s Health Initiative found that combination estrogen and progesterone hormone replacement therapy increases breast cancer and cardiovascular disease risk, which compelled many women to seek herbal alternatives such as black cohosh extract (BCE) to relieve their menopausal symptoms. While several clinical trials document the efficacy of BCE in alleviating menopausal symptoms, preclinical studies to determine how BCE works have yielded conflicting results. Part of this is because there is not a universally accepted method to standardize the dose of black cohosh triterpenes, the presumed active ingredients in the extract. Although the mechanism by which BCE relieves symptoms is unknown, several hypotheses have been proposed: it acts 1) as a selective estrogen receptor modulator, 2) through serotonergic pathways, 3) as an antioxidant, or 4) on inflammatory pathways. We found that while the most prominent triterpene in BCE, 23-epi-26-deoxyactein, suppresses cytokine-induced nitric oxide production in brain microglial cells, the whole BCE extract actually enhanced this pathway. A variety of activities have been reported for black cohosh and its compounds, but the absorption and tissue distribution of these compounds is unknown.Background: Omega-3 (n-3) polyunsaturated fatty acids (PUFAs) have modulating effects in several chronic inflammatory conditions. The aim of the present study was to test whether prior short-term dietary supplementation with n-3 (fish or seal oil) or n-6 (soy oil) PUFA rich oils would protect the development of dextran sulfate sodium (DSS)-induced colitis in rats. Methods: Forty-eight male Wistar rats were divided into 6 groups: no intervention, sham, DSS, seal oil + DSS, fi sh oil +DSS and soy oil + DSS. Following 7 days of acclimatisation, 1 mL oil (seal, fish or soy) or distilled water (sham) was administered by gavage day 8 to 14. Colitis was induced by 5% DSS in drinking water from day 15 to 21. Rats were sacrificed on day 23. Histological colitis (crypt and inflammation) scores, faecal granulocyte marker protein (GMP) and quantitative fatty acid composition in red blood cells were measured. Results: Pretreatment with fish or seal oils did not significantly influence DSS induced inflammation. In fact, all the oils tended to exacerbate the inflammation. Soy oil increased the mean crypt score (P < 0.04), but not the inflammation score or GMP. The ratio of n-6 to n-3 fatty acids (FAs) was 11 to 1 and 10 to 1 in standard diet and in red blood cells of control rats, respectively. Following administration of DSS, the ratio fell in all treatment groups (P < 0.001). The lowest ratios were seen in the groups receiving DSS + fi sh or seal oils (around 6 to 1). Conclusion: Short-term pretreatment with fish or seal oils did not protect against subsequent induction of colitis by DSS in this rat model. Whether the high ratio of n-6 to n-3 FAs in the standard diet concealed effects of n-3 FA supplementation should be further investigated.This feasibility study investigated the effects of Tai Chi, a mind-body exercise, on management of Type 2 diabetes mellitus. A total of 25 subjects (20-70 years) were recruited to participate in two 60-minute instructed Tai Chi exercise sessions each week for 12 weeks. The primary outcome measures (physiological variables) were hemoglobin A1C (HbA1c) taken at baseline and after 12 weeks of intervention, and self-reported fasting blood glucose level measured at baseline, 3, 6, 9, and 12 weeks of intervention. The secondary outcome measures (psychosocial variables) were Diabetes Quality of Life Questionnaire (Diabetes-39) and Exercise Self-Effi cacy administered at baseline and 12 weeks. A semi-structured interview was conducted at the end of the study (week 12). Paired t-tests was employed to determine all pre- and postintervention measurement changes, while individual growth curves were generated to show changes in fasting blood glucose levels during the study period. A rather high attrition rate of 48% was observed among the participants. The data showed no signifi cant effect of Tai Chi on HbA1c and self-reported fasting blood glucose, although there seemed to be a trend of lowered HbA1c. Analysis of subjects’ response suggested a positive experience for those who completed the intervention.In cancer treatment, apart from studying the effectiveness of chemo or radiotherapy in killing cancer cells, studies should examine ways of reducing drug side effects on patients and ways of enhancing the bodies’ immune system at the same time. Our defence system not only includes immune response, there are also detoxifying enzymes, antioxidant mechanisms, the ability for DNA damage repair and regulation of the hormone metabolism. Harmful environmental oestrogens that enter the human body can cause an increase of 16-α-hydroxyestrone as a harmful estradiol metabolite, the ratio between 16-α-hydroxyestrone and 2-hydroxyestrone relates to the risk of breast cancer. It is suggested that choosing nutritional products (that decrease the amount of 16-α-hydroxyestrone to regulate the hormone metabolism) can help with prevention of breast cancer. Increasing the ratio of monounsaturated fatty acid omega-3 (Ω-3) benefits health. Unsaturated fatty acid omega-6 (Ω-6) appears to be easily oxidised which can lead to DNA damage and increase the occurrence of cancer. The most important aspect to this approach is to reduce the ratio between saturated fatty acid and polyunsaturated fatty acid Ω-6, which is harmful to health. Olive oil has a high content of Ω-3 that benefits health. Ω-3 fatty acid can also be obtained from some fish, green vegetables and nuts. Linoleic acid is the most important source of Ω-6 fatty acid. Linolenic acid is the most important source of Ω-3 fatty acid. Natural foods e.g., purslane, is rich in Ω-3; the mustard family vegetables can increase the activity of detoxifying enzymes. Chinese Kiwi fruit drink reduces the side effects of the chemotherapy drug cyclophosphamide, which is also a DNA damaging agent. Soybean, job’s tears, garlic, mushroom varieties and tea have anti-cancer effects. Properly used nutritional products may assist treatment and recovery. Good balanced nutrition is essential for cancer healing.Spirituality seems to be an important cultural factor for African American women when thinking about their health. It is, however, not clear how spiritual health locus of control (SLOC) impacts health-related outcomes in the context of health message processing models, such as the Extended Parallel Process and the Risk Perception Attitude framework. Using a survey of African American and Caucasian women in the context of breast cancer, the role of SLOC and its interactions with perceived efficacy and risk was examined on four health outcomes—message acceptance, talking about breast cancer, information seeking, and behavioral intentions. For African American women, SLOC had a positive impact for talking about breast cancer through an interaction with risk and efficacy such that women high in both SLOC and perceived efficacy, but low in perceived risk were more likely to talk about breast cancer than when efficacy was low. However, high SLOC exacerbated the negative effects of efficacy on talking about breast cancer regardless of the risk level for Caucasian women. SLOC also had a positive influence on attending to breast cancer information in the media for African American women. SLOC played no role in attending to breast cancer information for Caucasian women. Interestingly, SLOC played no role for African American women on behavioral intentions, however, it worked to decrease behavioral intentions for Caucasian women when risk was high.