In vitro effect of important herbal active constituents on human cytochrome P450 1A2 (CYP1A2) activity

This study was designed to investigate eight herbal active constituents (andrographolide, asiaticoside, asiatic acid, madecassic acid, eupatorin, sinensetin, caffeic acid, and rosmarinic acid) on their potential inhibitory effects on human cytochrome P450 1A2 (CYP1A2) activity. A fluorescence-based enzyme assay was performed by co-incubating human cDNA-expressed CYP1A2 with its selective probe substrate, 3-cyano-7-ethoxycoumarin (CEC), in the absence or presence of various concentrations of herbal active constituents. The metabolite (cyano-hydroxycoumarin) formed was subsequently measured in order to obtain IC₅₀ values. The results indicated that only eupatorin and sinensetin moderately inhibited CYP1A2 with IC₅₀ values of 50.8 and 40.2 μM, while the other active compounds did not significantly affect CYP1A2 activity with IC₅₀ values more than 100 μM. K_i values further determined for eupatorin and sinensetin were 46.4 and 35.2 μM, respectively. Our data indicated that most of the investigated herbal constituents have negligible CYP1A2 inhibitory effect. In vivo studies however may be warranted to ascertain the inhibitory effect of eupatorin and sinensetin on CYP1A2 activity in clinical situations     

Attenuation of abnormal glutamate release in zinc deficiency by zinc and Yokukansan

The mechanism of the abnormal increase in extracellular glutamate concentration in the hippocampus induced with 100 mM KCl in zinc deficiency is unknown. In the present study, the changes in glutamate release (exocytosis) and GLT-1, a glial glutamate transporter, expression were studied in young rats fed a zinc-deficient diet for 4 weeks. Exocytosis at mossy fiber boutons was enhanced as reported previously and GLT-1 protein was increased in the hippocampus. The enhanced exocytosis is thought to increase extracellular glutamate concentration. However, the basal concentration of extracellular glutamate in the hippocampus was not increased by zinc deficiency, suggesting that GLT-1 protein increased serves to maintain the basal concentration of extracellular glutamate. The enhanced exocytosis was attenuated in the presence of 100 μM ZnCl₂, which attenuated the abnormal increase in extracellular glutamate induced with high K⁺ in zinc deficiency. The present study indicates that zinc attenuates abnormal glutamate release in zinc deficiency. The enhanced exocytosis was also attenuated in slices from zinc-deficient rats administered Yokukansan, a herbal medicine, in which the abnormal increase in extracellular glutamate induced with high K⁺ was attenuated. It is likely that Yokukansan is useful for prevention or cure of abnormal glutamate release. The enhanced exocytosis in zinc deficiency is a possible mechanism on abnormal increase in extracellular glutamate in the hippocampus induced with high K⁺.     

Applications of LC-MS in the study of the uptake, distribution, metabolism and excretion of bioactive polyphenols from dietary supplements

Specific and quantitative analyses of the bioactive components and their metabolites in body fluids are essential to assess the interaction between groups of compounds in dietary supplements and preparations of psychoactives. Reverse-phase LC separations combined with tandem mass spectrometry provide the necessary specificity and sensitivity. In this paper, applications of these methods are described for the analysis of isoflavones, salvinorin A, synephrine isomers and their metabolites in serum, urine and aqueous humor.     

Association of genetic variants with response to iron supplements in pregnancy

The incidence of iron deficiency anemia in pregnancy is high in India where iron supplementation is a regular practice. The response to oral iron is influenced by several factors such as age, body mass index, gravida, socioeconomic status, food, vitamin deficiency and compliance to supplements. The major challenge is to understand the various modulators of iron status in this high-risk group so that we can improve the diagnosis and the management of these patients. The current study was designed to evaluate the iron status during pregnancy and to identify factors which might be influencing their response to oral iron. We investigated a total of 181 pregnant women with anemia (Hb < 11 g/dl) and evaluated the impact of probable factors on anemia and their iron status. Assessment of the response was based on hemoglobin and serum ferritin or transferrin saturation level after 8 and 20 weeks of iron supplementation. Socioeconomic, clinical, hematological, biochemical and genetic factors were all evaluated. Molecular analysis revealed that HFE variant allele (G) (rs1799945) was significantly associated with an adequate response to iron supplementation. We identified five subjects with a sustained poor response, and targeted re-sequencing of eleven iron-related genes was performed in them. We have identified seven novel variants in them, and in silico analysis suggested that these variants may have an iron regulatory effect. Taken together, our findings underscore the association of genetic variants with response to supplements in pregnancy, and they can be extended to other diseases where anemia and iron deficiency coexist.

Electronic supplementary material

The online version of this article (doi:10.1007/s12263-015-0474-2) contains supplementary material, which is available to authorized users.     

Clinical and Analytical Toxicology of Dietary Supplements: A Case Study and a Review of the Literature

The use of dietary supplements has grown dramatically in the last decade. A large number of dietary and herbal supplements escape regulatory and quality control; components of these preparations are poisonous and may contain, among other toxins, heavy metals. Uncontrolled use of dietary and herbal supplements by special populations, such as the military, may therefore pose a health risk. Clinical symptoms are not always properly attributed to dietary supplements; patients often do not mention supplement use to their health care provider. Therefore, a health risk estimate is hard to make on either the individual or the population level. The literature on this issue was reviewed and discussed in the light of a representative clinical-chemical case study. This case study was performed on a host of preparations that were used by one single individual in the military. Both essential (chromium, copper, zinc, and iron) and poisonous (arsenic, lead, and nickel) trace elements were determined using inductively coupled plasma combined with optical emission spectrometry (ICP-OES) or with mass spectrometry (ICP-MS). Arsenic and lead were detected at exposure levels associated with health risks. These health risks were detected predominantly in hormone-containing supplements and the herbs and botanicals used for performance enhancement. To the extent that this is a representative sample, there is an underestimation of supplement use and supplement risk in the US military, if not in the general population. Since clinical symptoms may be attributed to other causes and, unless patients are specifically asked, health care providers may not be aware of their patients' use of dietary supplements, a strong support of laboratory diagnostics, such as a toxicological screening of blood or urine, is required. In addition, screening of the preparations themselves may be advised.     

Encapsulation and modified-release of thymol from oral microparticles as adjuvant or substitute to current medications

The aim of this study was to encapsulate, thymol, in natural polymers in order to obtain (i) taste masking effect and, then, enhancing its palatability and (ii) two formulations for systemic and local delivery of herbal drug as adjuvants or substitutes to current medications to prevent and treat several human and animal diseases. Microspheres based on methylcellulose or hydroxypropyl methylcellulose phthalate (HPMCP) were prepared by spray drying technique. Microparticles were in vitro characterized in terms of yield of production, drug content and encapsulation efficiency, particle size, morphology and drug release. Both formulations were in vivo orally administered and pharmacokinetic analysis was carried out. The polymers used affect the release and, then, the pharmacokinetic profile of thymol. Encapsulation into methylcellulose microspheres leads to short half/life but bioavailability remarkably increases compared to the free thymol. In contrast, enteric formulation based on HPMCP shows very limited systemic absorption. These formulations could be proposed as alternative or adjuvants for controlling pathogen infections in human or animal. In particular, methylcellulose microspheres can be used for thymol systemic administration at low doses and HPMCP particles for local treatment of intestinal infections.     

Immunomodulation property of hexane fraction of leaves of Cinnamomum tamala Linn. in rats

The leaves of Cinnamomum tamala Linn. (CT) (Lauraceae) clinically used in Ayurveda as antidiabetic and diuretic, but no reports are available towards immunomodulating property. Its hexane fraction (CTH) was orally given to rats for 10 days and delayed type of hypersensitivity (DTH), antibody producton against sheep red blood cells (SRBCs), mitotic index in bone marrow cells and concanavalin A (Con A) mediated proliferation of lymphocytes were assessed. Further on 30 days treatment, change in body weight (BW), spleen weight, thymus weight, bone marrow cellularity and hematological changes were observed. It inhibited significantly the DTH response (IC₅₀ 1475 ± 57.19 mg kg^?1 BW), antibody production, suppressed mitotic index in bone marrow cells along with the suppression of lymphocyte proliferation against Con A (IC₅₀ 63.33 ± 1.95 µg mL^?1). In all experiments, cyclophasphamide and dexamethasone had been used as reference drug for in vivo and in vitro studies, respectively. On 30 days treatment, the CTH (800 mg kg^?1 BW and above) significantly suppressed growth rate, increase of spleen and thymus weight and low bone marrow cellularity. In hematological examination, it inhibited total white blood cell and lymphocytes count and increased per cent of polymorphs. Thus, it could be suggested that the fraction possesses immunosuppressive property at doses, higher than 800 mg kg^?1 BW in rats. Copyright © 2010 John Wiley & Sons, Ltd.     

Chitosan Supplementation and Fecal Fat Excretion in Men

Objective : Few weight loss supplements are clinically tested for efficacy, yet their proliferation continues. Chitosan‐based supplements are sold as fat trappers and fat magnets. They purportedly block fat absorption and cause weight loss without food restriction. We quantified the in vivo effect of a chitosan product on fat absorption. Research Methods and Procedures : Participants (n = 15) consumed five meals per day for 12 days. Energy intake was not restricted. Participants consumed no supplements during a 4‐day control period and two capsules five times per day (4.5 g chitosan/d), 30 minutes before each meal, during a 4‐day supplement period. All feces were collected from days 2 to 12. Oral charcoal markers permitted division of the feces into two periods. The two fecal pools were analyzed for fat content. Results : Participants were male, 26.3 ± 5.9 years old, BMI of 25.6 ± 2.3 kg/m². Subjects consumed 133 ± 23 g of fat/d and 12.91 ± 1.79 MJ/d (3084 ± 427 kcal/d). Individual meals averaged 26.3 ± 9.3 g of fat. With chitosan supplementation at 10 capsules/day, fecal fat excretion increased by 1.1 ± 1.8 g/d (p = 0.02), from 6.1 ± 1.2 to 7.2 ± 1.8 g/d. Discussion : The effect of chitosan on fat absorption is clinically negligible. Far from being a fat trapper, at 0.11 ± 0.18 g of fat trapped per 0.45‐g capsule or 1.1 g (9.9 kcal) fat trapped per day, this product would have no significant effect on energy balance. The fat trapping claims associated with chitosan are unsubstantiated.     

益气活血中药复方对CVB3大鼠心肌细胞感染模型ATP6基因表达的影响

目的：研究益气活血中药复方对CVB3大鼠心肌细胞感染模型ATP6（ATP synthase F0 subunit 6）基因表达的作用机制。方法：本实验用新生2-3dWistar大鼠心肌细胞,建立CVB,病毒感染模型,通过改良的抑制性消减杂交技术（SSH）,克隆了受CVB,攻击的心肌细胞中被中药（益气活血中药复方）调控的基因。结果：ATP6基因,在中药组中高表达,而病毒组中表达减弱或抑制。结论：益气活血中药复方能影响受CVB,病毒攻击的宿主细胞ATP6基因的表达,有效保护心肌、阻断病程进度,从而实现治疗病毒性心肌炎的目的。     

Nutrient intakes in women and risks of anophthalmia and microphthalmia in their offspring

BACKGROUND: There is a paucity of information about risk factors for the human eye anomalies anophthalmia and microphthalmia. In this population-based case-control study we investigated whether periconceptional intakes of supplemental folic acid, dietary folate, vitamin A, and several other nutrients were associated with these eye defects. METHODS: This study included data on deliveries that had estimated due dates from 1997-2002 and were part of the National Birth Defects Prevention Study (the National Birth Defects Prevention Study is a population-based case-control study of a wide spectrum of birth defects, incorporating data from 10 birth defects surveillance systems in the United States [Arkansas, California, Georgia/Centers for Disease Control and Prevention, Iowa, Massachusetts, New Jersey, New York, North Carolina, Texas, and Utah]). Cases were those infants or fetuses born with either anophthalmia or microphthalmia. Liveborn infants without major malformations were eligible as controls. Maternal interviews were conducted, primarily by telephone, in English or Spanish. Participation in the interview was 71% among case mothers and 68% among control mothers. Interviews were completed with 89 case mothers and 4,143 control mothers. A shortened version of the food frequency questionnaire from the Nurse's Health Study was used to assess frequency of intake of 58 food items during the year before pregnancy. RESULTS: Our results did not indicate reduced risks for these eye malformations associated with maternal intake of vitamin supplements containing folic acid. The data did not show an association between malformation risk and higher or lower intakes of vitamin A. We also did not observe strong evidence that an abundance or a lack of dietary intake of any other nutrient was associated with increased risk of the studied eye malformations. CONCLUSIONS: Our observations contribute to a limited body of findings on these rare eye defects.