PREVALENCE OF 2756A>G POLYMORPHISM OF METHIONINE SYNTHASE GENE IN POPULATIONS OF EASTERN SIBERIA

This aim of the study was to research the prevalence of genotypes and alleles of2756A>G polymorphism of gene of methionine synthase (MTR) gene in Russian and Buryat populations of Eastern Siberia. We used DNA blood database of children and teenagers of the Russian and Buryat nationality living in Eastern Siberia. Genotyping was carried out using standard PCR method. Prevalence of minor G-allele in Russian population was 25,7 %, in Buryat population - 13,2 %. We revealed differences in prevalence of genotypes and alleles between studied groups (p = 0,051; p = 0,032)

Q223R polymorphism of the LEPR and obesity

The problem of overweight and obesity is one of the most urgent health issues in the world. 13 % of girls and 21 % of boys aged 11 suffer from overweight in the Russian Federation. The main causes of pubertal obesity are endocrine pathology, lifestyle and genetic disorders including mutation and polymorphisms of different metabolic pathways. Leptin produced in adipose tissue participates in reproduction regulation, glucose homeostasis, bone formation, etc. These effects are provided by leptin receptors coding LEPR gene. Q223R (rs1137101) polymorphism is associated with an increased serum level of leptin and overweight. There is no exact information about association between this polymorphism and obesity of adolescent females. The objective was to reveal LEPR Q223R polymorphism association between overweight and obesity in adolescent females. 123 Caucasian adolescent females were involved in this study. All samples could be separated into two groups: the girls with normal weight (SDS BM1 ± 1.0; control group), girls with overweight and obesity (SDS BM1 > +1.0-2.0; studied group). Anthropometric measurements (weight, height, waist and hip circumference, body fat percentage) were taken, and genotyping was performed using polymerase chain reaction with electrophoresis detection. G-allele frequency was 43.1 % in control and 40 % in the clinical group. We found no significant differences of the prevalence of polymorphism Q223R between the studied groups (р = 0,862). Furthermore, there was no association between the carriage of AG and GG with weight, BM1, body fat percentage, waist and hip circumference in both groups (р > 0.05). We have not found any association between LEPR Q223R and overweight and obesity in adolescent females

Gene of folate cycle MTHFR and nutrition

Climatic and geographical features always influenced on adaptation to the environment as well as the development level of economy and culture. The formation of certain food behavior in population determined fixation of specific gene alleles and mutation. They were responsible for digestion a typical food for the population. At the moment there is information about such genetic features as metabolism of proteins, carbohydrates and fats for many races and nations. But a few studies are about causes of prevalence differences of gene polymorphisms of folate cycle in the world's populations. The main objective was to explore 677C>T mutation MTHFR gene among Russian and Buryat to compare it with ones in other populations. The total of399 children and adolescents (200 Russians and 199 Buryats) were involved in this study. Genotyping of MTHFR 677C>T was performed by PCR-RT We used soft "STATISTICA 8.0" to compare results. As a result, T-allele frequency was 30 % in Russians and 21,1 % in Buryats. We found significant differences of prevalence of polymorphism 677C>T between studied groups (p = 0.011). Also we found significant difference between Russian and British (p = 0.054), French (p = 0.0263), Spanish (p < 0.0001), Italian (p < 0.0001) and Greek samples (p = 0.0454). The Buryat group had significant differences with Chinese (p < 0.0001), Korean (p < 0.0001), Mongolian (p < 0.004), Japanese (p < 0.0001), Kazakh (p = 0.0198) samples and two samples of Hans (p < 0.0001; p = 0.0390)

Metabolism and obesity: role of leptin receptor gene

Background. Currently more than 119 obesity-related polymorphisms is known to participate in adult obesity. One of them is LEPR Q223R. Many researches shown association of this polymorphism with adult obesity. However, the role of LEPR Q223R in adolescent overweight and obesity is the matter of dispute. Aim: to determine association of polymorphism Q223R of LEPR gene with some biochemical and hormonal measurements of blood in female adolescents with normal weight and with overweight and obesity. Materials and methods. A total of 103 female adolescents (14-17 years of age) was examined. All girls were divided into 2 groups: 43 girls with normal weight (SDS BM 10.311 ± 0.585), and 65 girls with overweight and obesity (SDS BMI 2.255± 0.739) (р < 0.0001). Height, weight, BM1, SDS BM1 were measured. Laboratory tests included triglycerides, total cholesterol and its fraction, TTH, free thyroxin and leptin. All girls were genotyped on carrier of LEPR Q223R. Statistical analysis was provided by software Statistica 8.0 using nonparametric Mann - Whitney methods and Chi-square test with Yates correction. Results. Significant association of carrying RR-genotype with increase of SDS BM1 (p = 0.006), THS (p = 0.006) and decrease of free thyroxin was shown in control group. Conclusion. Our results showed the association of R-allele with increase of SDS BM1, THS and decrease T4 free in control group

Beyond bone biology: Lessons from team science

Today, research in biomedicine often requires the knowledge and technologies in diverse fields. Therefore, there is an increasing need for collaborative team science that crosses traditional disciplines. Here, we discuss our own lessons from both interdisciplinary and transdisciplinary teams, which ultimately ushered us to expand our research realm beyond bone biology. This article is protected by copyright. All rights reserved

FREQUENCIES OF POLYMORPHISMS IN THE CYTOCHROME’S P450 GENES OF WARFARIN TRANSFORMATION IN A EUROPEAN POPULATION OF EASTERN SIBERIA

Background. Genotypes of the cytochrome p450 isoform (CYP2C9 and CYP4F2) determine warfarin dose requirements. Frequencies of risk alleles and genotypes of CYP2C9 and CYP4F2 gene vary in different races and ethnic groups.Aim. This study analyzed the frequencies of *2, *3 alleles of CYP2C9 gene and the 1347 C&gt;T allele of CYP4F2 gene in the Caucasians of Eastern Siberia, and compare with other populations.Materials and methods. Participants were 147 patients (Caucasians): 67 (45.58 %) man and 80 (54.42 %) women), taking warfarin for the prevention of thrombosis with a mean age of 64.74 ± 14.29 years. There were patients with atrial fibrillation – 77 (52.38 %) persons, coronary artery disease – 10 (6.80 %), pulmonary embolism – 5 (3.40 %), 15 (10.20 %) patients after implantation of an mechanical heart valve, etc. The subjects were genotyped for CYP2C9 (*1,*2,*3), and CYP4F2 (1347 C&gt;T) by real-time polymerase chain reaction (RT-PCR) using “Pharmacogenetics Warfarin” reagent kits (DNA technology, Russia).Results. 69.4 % of Caucasians of Eastern Siberia (Russians), have two functional alleles (*1/*1) of CYP2C9 (they’re extensive/normal metabolizers), the number of intermediate metabolizers (*1/*2, *1/*3) was 29.8 % and 0.68 % of slow metabolizers (*3/*3). Homozygous carriers of two non-functional alleles *2 and *3 (*2/*2, *2/*3) were absent. Carriers of one coumarin-resistant Т-allele of CYP4F2 were 57 (38.7 %) respondents, two coumarin-resistant alleles – 10 (6.8 %) respondents.Conclusions. Frequencies of polymorphisms in the Cytochrome’s p450 genes of warfarin transformation in a European population of Eastern Siberia have no differences with other European populations of the worl

The experience in the development and validation of method for testosterone measurement in blood serum of premenopausal women using HPLC-MS/MS

Testosterone assessment is essential for detecting biochemical hyperandrogenism, one of the important diagnostic criteria of polycystic ovary syndrome (PCOS) both in clinical practice and in epidemiological studies. Currently, tandem liquid chromatography-mass spectrometry (LC-MS/MS) is the most preferred technique to measure testosterone level in women. Its validation is important to reproducibility of androgen tests results for clinical practice and for epidemiological studies of the prevalence PCOS.The aim of the study. To develop and validate a method for determining total testosterone in blood serum using highly efficient LC-MS/MS to assess androgenemia in the epidemiological study of the prevalence of PCOS and its phenotypes in Eastern Siberia (ESPEP STUDY).Materials and methods. We determined a total testosterone level in serum blood using triple quadrupole mass spectrometer LCMS-8060 (Shimadzu, Japan). The protocol of technique was developed using self-prepared purified human testosteronefree serum with a known concentration of analyzed compound. We used the serum samples of women of reproductive age to test the developed method.Results. Optimum chromatographic conditions were obtained with a Kromasil 100-2.5-C18 column (2.1 mm × 100 mm; AkzoNobel, Netherlands), and an isocratic elution mode using a mobile phase consisting of acetonitrile and 0.1 % aqueous solution of formic acid. The total flow rate was 0.35 ml/min. The lower limit of quantification was 5 ng/dl with an average accuracy of 100.2 %. During the approbation of the method in a test population sample of 1138 premenopausal women (mean age – 34.3 ± 6.3 years), the median testosterone concentration was 26.9 ng/dl.Conclusion. It was found that the proposed method for determining testosterone in blood serum has acceptable linearity and reproducibility and meets the requirements for bioanalytical methods under the regulatory documentation. This method can be used for clinical practice and epidemiological study of the prevalence of PCOS

First-in-class humanized FSH blocking antibody targets bone and fat

Blocking the action of FSH genetically or pharmacologically in mice reduces body fat, lowers serum cholesterol, and increases bone mass, making an anti-FSH agent a potential therapeutic for three global epidemics: obesity, osteoporosis, and hypercholesterolemia. Here, we report the generation, structure, and function of a first-in-class, fully humanized, epitope-specific FSH blocking antibody with a KD of 7 nM. Protein thermal shift, molecular dynamics, and fine mapping of the FSH-FSH receptor interface confirm stable binding of the Fab domain to two of five receptor-interacting residues of the FSHβ subunit, which is sufficient to block its interaction with the FSH receptor. In doing so, the humanized antibody profoundly inhibited FSH action in cell-based assays, a prelude to further preclinical and clinical testing

ПРЕДИКТИВНЫЕ МАРКЕРЫ ОТВЕТА НА БЛОКАТОРЫ КОНТРОЛЬНЫХ ТОЧЕК ИММУННОГО ОТВЕТА

Despite the unprecedented success in using immune checkpoint inhibitors in the treatment of lung cancer, melanoma, hypermutable tumors of various localization, etc., a significant proportion of patients receiving these drugs do not respond to treatment. Predictive markers routinely used in the selection of patients for immunotherapy, in particular, the level of expression of PD -L1 and the presence of microsatellite instability, have certain limitations. Over the past decade, many other biomarkers designed to predict response to immunotherapy have been proposed, namely: tymor mutation burden, composition of lymphocytic infiltrate; allelic composition of the major histocompatibility complex; relationship between the numbers of different formed elements of blood as well as between its biochemical parameters; microflora of the digestive tract, etc. These markers can directly or indirectly reflect the immunogenicity of the tumor itself, as well as the state of systemic and intratumoral immune response. The predictive power and reliability of these markers are extremely different. When preparing this review, we conducted a literature search for recent studies regarding predictors of efficacy for immune checkpoint inhibitors published in the journals included in the databases, such as Pubmed, Web of Science, and Scopus.Несмотря на беспрецедентный успех применения ингибиторов контрольных точек иммунного ответа в терапии рака легкого, меланомы, гипермутабельных новообразований различной локализации и проч., существенная доля пациентов, получающих препараты этой группы, не демонстрирует ответа на лечение. Предиктивные маркеры, рутинно используемые при отборе больных для прохождения иммунотерапии, в частности уровень экспрессии PD -L1 и наличие микросателлитной нестабильности, имеют определённые ограничения. За последнее десятилетие было предложено множество иных биомаркеров, призванных предсказать эффект на назначение иммунотерапии: внутриопухолевая мутационная нагрузка, состав лимфоцитарного инфильтрата; аллельная композиция главного комплекса гистосовместимости; соотношения между количеством различных форменных элементов крови и ее биохимическими показателями; особенности микрофлоры ЖКТ и т. д. Эти маркеры могут прямо или косвенно отражать иммуногенность самой опухоли, а также состояние системного и внутриопухолевого иммунитета. Предсказательная сила, достоверность и степень их близости к практическому применению чрезвычайно различны. При подготовке данного обзора был проведен поиск литературы последних лет, касающейся изучения предикторов эффективности ингибиторов контрольных точек иммунного ответа, в журналах, входящих в базы данных Pubmed, Web of Science, Scopus