Urinary excretion profile of higenamine in females after oral administration of supplements - Doping scenario.

In 2017, higenamine was added to the World Antidoping Agency's (WADA) Prohibited list under group S3: beta-2 agonists and it is banned for athletes both in - and out of competition. Aim of this study was to characterize the urinary excretion profile of higenamine and its metabolite coclaurine after oral administration of multiple doses of higenamine capsules. For this purpose, an administration study including female basketball players was performed. For the detection of higenamine and cocalurine in the collected urine samples, a new, fast, and highly sensitive quantitative on-line SPE LC HRMS method was developed and validated. The method was applied for the quantification of higenamine and cocalurine in urine and their excretion pattern was defined. Results obtained show substantial inter-individual differences in the excretion profile of higenamine and coclaurine. For higenamine, half-lives were estimated to be between 4 and 27 h, and for coclaurine between 5 and 25 h. Furthermore, the data indicate that the elimination of coclaurine is rate-limited by its formation. Higenamine could be detected at a urine concentration above 10 ng/mL for at least 20 h after the last application for all study participants.

Xanthine oxidase activity in patients with age-related cataract associated with hypertension.

Reactive oxygen species and lipid peroxidation are important factors that contribute to the development of age-related cataract. The study included 130 patients with age-related cataract, 69 of whom were diagnosed with hypertension (HT), 20 with hypertension and type 2 diabetes mellitus (DM), and 41 had no accompanying condition. The following parameters were measured in the serum of the examinees: products of lipid peroxidation malondialdehyde (MDA) and lipofuscin-like fluorophores (LLF), activity of prooxidative enzymes xanthine oxidase (XO) and myeloperoxidase (MPO), antioxidant enzymes superoxide dismutase (SOD) and glutathione peroxidase (GPx), the concentration of thiol groups, and the ferric reducing activity of plasma. The activity of prooxidative enzymes XO and MPO was higher in the plasma of patients with HT (XO=9.0±1.2 U/L; MPO=77.3±8.4 U/L) and with HT and DM (XO=11.9±0.9 U/L; MPO=89.5±5.0 U/L) compared to patients with age-related cataract (XO=6.2±0.9 U/L; MPO=52.4±6.3 U/L; P<0.01). Our research has shown that patients with age-related cataract and hypertension were exposed to increased oxidative damage of biomolecules, based on the increased plasma LLF and MDA content and decreased levels of thiol groups. Oxidative changes of biomolecules in these patients were associated with increased activity of the XO, MPO, and GPx enzymes and a lower extracellular SOD activity and total ferric reductive ability of plasma.

Influence of pharmacological education on perceptions, attitudes and use of dietary supplements by medical students.

BACKGROUND: The ready availability and use of dietary supplements (DS) by the public means that healthcare professionals require education in this area. In the Republic of Serbia, education related to use of DS is included in undergraduate medical training and it is therefore important to assess the effectiveness of this education. The aim of our survey was to investigate the influence of pharmacological education on the use, attitudes and perceptions of risks associated with DS among medical students. METHODS: Medical students at the University of Kosovska Mitrovica participated in the survey. Three hundred eighty questionnaires were distributed, yielding a response rate of 89% (n = 334). Data were categorized by year of study, completion of a one-year course in pharmacology and having passed the final exam. The results were compared between 192 (58%) medical students educated in pharmacology (MSEP) and 142 (42%) medical students not educated in pharmacology (MSNEP). The questionnaire was divided into 4 parts: socio-demographic and lifestyle/behavioral characteristics, use of DS, attitudes about efficacy, safety and perception of risk due to DS use. Chi-square test, Student's t-test, and Mann-Whitney U test were used for statistical analysis. RESULTS: About 53% of respondents used some form of DS. Attitudes regarding the safety of DS consumption showed a difference between the groups. MSEP were more likely to agree that DS have the potential to cause adverse reactions (Likert scale mean 4.1 vs. 3.5, p < 0.001) as well as interactions with conventional drugs (Likert scale mean 4.2 vs. 3.2, p < 0.001) than MSNEP. Finally, MSEP ranked St. John's wort and ginkgo as the most dangerous DS, but creatine and vitamin C were both ranked as relatively safe. Conversely, MSNEP considered ginkgo and vitamin C the most harmful DS, claiming that omega-3 fatty acids and vitamin D had the least hazardous side effects. CONCLUSION: Our results showed that pharmacological education gives young medical students a better understanding of the risks of DS-drug interactions and potential adverse effects. However, their overall attitudes and perception of risk indicate the need for further education.

Determination of higenamine in dietary supplements by UHPLC/MS/MS method.

From 1st January 2017 higenamine was added on the WADA (World Anti-doping Agency) Prohibited list under S3 group beta-2 agonists as at all times banned substance for the athletes. The main origine of higenamine (or norcoclaurine) are different plants including Nandina domestica, Aconitum carmichaelii, Asarum heterotropioides, Galium divaricatum, Annona squamosa, Nelumbo nucifera etc. Higenamine main use is related to weight loss and it could be found (un)labeled in different dietary supplements. The objective of this study was development of sensitive and reliable UHPLC/MS/MS method for determination of higenamine in various dietary supplement samples. In order to obtain high method sensitivity, hydrophilic interaction liquid chromatography (HILIC) mode was applied. Separation was carried out on UHPLC Acquity BEH HILIC analytical column (2.1mm×100mm, 1.7µm particle size). Mobile phase consisted of 0.1% formic acid in water and acetonitrile, respectively, was mixed in ratio of 30:70, v/v. Flow rate was set at 0.2mLmin-1. Quercetin was used as an internal standard. ESI (+) source ionization mode using multi reaction monitoring (MRM) mode was utilized and three ion transitions of higenamine were followed 272.08→107.01, 272.08→161.07 and 272.08→77.08. Developed method was fully validated and applied for identification and quantification of higenamine in different dietary supplements. According to the results, the most of investigated supplements were free of higenamine, and on the other hand, presence of higenamine was confirmed in some samples while it was not declared on the label.

[Therapy of Helicobacter pylori infection using Lactobacillus acidophilus]. / Terapija Helicobacter pylori infekcije primenom Lactobacillusa acidohpilus.

INTRODUCTION: In vitro experiments with Lactobacillus acidophilus have revealed its inhibitory effect on Helicobacter pylori and its application in treatment of Helicobacter pylori positive gastritis was examined. MATERIAL AND METHODS: 15 patients have undergone gastroscopy with biopsy and by histopathological examination. Helicobacter pylori positive gastritis was detected. During a two-month period these patients took acidophilus milk (3 x 250 ml a day) prepared according to a special protocol and which contained 4 x 10(9)-1 x 10(10) live cells of Lactobacillus acidophilus at the moment of preparation. Lactobacillus acidophilus strain NAS, gained from lyophilized preparation Bio-Nate (Natren Inc. USA) was used as a test organism. Control gastroscopy was performed 2 months later. RESULTS: 14 patients have completed the examination. All of them were satisfied with the taste of acidophilus milk and could stand it well, whereas in 6 out of 14 Helicobacter pylori was eradicated. DISCUSSION: Helicobacter pylori eradication therapy is recommended in all cases of Helicobacter pylori positive gastritis associated with peptic ulcer, as well as in absence of ulcer when subjective difficulties occur. Antibiotic therapy is often unsuccessful and most often associated with risks of significant adverse effects, being the consequence of intestinal microflora disorders. The aim of using Lactobacillus acidophilus in the therapy is to reduce risks of adverse effects. In our study, by using acidophilus milk only, without other therapy, eradication of Helicobacter pylori was achieved in 6 out of 14 patients. All patients could stand the therapy well and were satisfied with the taste of the preparation. The number of examinees was small in regard to making conclusions, but the results are encouraging and show that apart from established in vitro effect. Lactobacillus acidophilus has a potential in vivo effect.

Lowering of ochratoxin A level in milk by yoghurt bacteria and bifidobacteria.

A decrease in the content of ochratoxin A (OA) was observed in milk samples fermented by yoghurt bacteria and bifidobacteria. OA was added to the milk before fermentation at a rate of 0.05-1.5 mg/L. No residues of OA were found in samples containing 0.05 and 0.1 mg/L of OA, fermented by S. Salivarius subsp. thermophilus, L. delbrueckii subsp. bulgaricus and B. bifidum. Yoghurt bacteria (S. salivarius subsp. thermophilus and L. delbrueckii subsp. bulgaricus) were the most effective since no residues were detected even in fermented samples containing originally 0.5 mg/L OA.

Decrease of aflatoxin B1 in yoghurt and acidified milks.

Fermentation of yoghurt and acidified milks containing aflatoxin B1 (AB1) were studied. AB1 added to milk before fermentation at concentrations of 600, 1000 and 1400 micrograms/kg was reduced in yoghurts (pH 4.0) by 97, 91 and 90%, respectively. Coagulation time was approximately the same as in the controls. Streptococci had longer chains than those in the controls. The main decrease of AB1 occurred during the milk fermentation. A decrease of AB1 (conc. 1000 micrograms/kg) in milks acidified with citric, lactic and acetic acids (pH 4.0) was 90, 84 and 73%, respectively.