Search

Relevance. Acute respiratory infections (ARI) are a serious health problem not only because of the high frequency of their occurrence, but also because of the economic damage they cause both in the form of direct costs (the cost of diagnosis and treatment) and indirect costs (disability, reduced labor productivity, etc.). Pregnant women and children under 5 years of age are included in the group of patients with risk factors for complications of influenza and other ARI, therefore, an analysis of the etiological structure of ARI and influenza in obstetric hospitals is an urgent task. In recent years, there has been an urgent need to create a national complex diagnostic test system based on molecular genetic methods for detecting infectious agents that cause ARI. Aims. The aim of the study is to analyze the etiological structure of ARI and influenza in patients with clinical symptoms and to develop and implement a new test system for rapid screening and diagnosis of infections that cause ARI. Materials & methods. When studying the etiological structure of ARI and influenza, cultural studies of the nasal and pharyngeal mucosa were carried out, followed by identification of microorganisms using MALDI-TOF mass spectrometry and molecular genetic study (real-time PCR) using an experimental test panel containing primers that allow detecting the following viruses: influenza A, B viruses, parainfluenza viruses of the 1st, 2nd, 3rd and 4th types, coronaviruses OS43, HKU1, NL63, E229, respiratory syncytial virus, metapneumovirus, rhinovirus and adenovirus, as well as bacterial pathogens of ARI: Haemophilus influenzae, Streptococcus pneumoniae, Streptococcus pyogenes, Moraxella catarrhalis, Staphylococcus aureus, Klebsiella pneumoniae, Pseudomonas aeruginosa. The test system was developed using the following methods: real-time PCR, a combination of reverse transcription and real-time PCR (RT-PCR) and the next generation sequencing (NGS) method. Results. The etiological structure of ARI and influenza was analyzed in patients with clinical manifestations (cough, tickling/sore throat/hyperemia of the mucous membrane of the palate and the back wall of the pharynx, shortness of breath/difficulty breathing, acute runny nose/nasal congestion). The species spectrum of bacterial and viral pathogens was revealed. A new test system based on PCR, real-time RT-PCR and NGS has been created for complex diagnostics of both viral and bacterial pathogens of ARI, consisting of three separate components: the main test system «ARI», which detects the main viral and bacterial pathogens of ARI, and two additional sets of reagents: «Oseltamivir resistance» and «Oseltamivir/ Zanamivir resistance». Conclusions. The new test system can be used to detect and differentiate nucleic acids of pathogens of ARI of humans. The test system seems to us promising for further use. As a result of the analysis of the etiological structure of acute respiratory infections and influenza, attention is drawn to a significantly smaller variety of identified pathogens in 2020 and a much more pronounced dominance of rhinovirus infection compared to our previous study in 2019.

Objective: to identify molecular genetic predictors of metastatic spread to regional lymph nodes in patients with breast cancer (BC) based on the analysis of gene expression profile of the primary tumor.Materials and methods. The study included 358 patients with BC who underwent surgical treatment in breast cancer department of Academician V.I. Kulakov Research Center of Obstetrics, Gynecology, and Perinatology, Ministry of Health of Russia. Among all included into the study patients, 132 (36.9 %) had metastases in at least one axillary lymph node. Molecular genetic examination of the tumor tissue was carried out using reverse transcription polymerase chain reaction; the diagnostic panel consisted of 45 functional and 3 reference genes. Results. Patients with metastases to regional lymph nodes were generally younger (p = 0.006), had larger primary tumor (pConclusion. TMEM45A, CCND1 and MIA gene expression in the primary tumor were the markers of lymph node involvement in BC. The developed predictive genetic signature can become an additional diagnostic tool to predict the risk of lymph node metastases at the point of planning the volume of axillary surgery in patients with BC.

Study Objective: To evaluate the effect of low molecular heparin (LMH) therapy on non-invasive prenatal DNA screening (NIPS) for chromosomal pathologies. Study Design: cross-sectional study. Materials and Methods. We have examined two groups of pregnant women: group I included 49 patients with constantly low foetal DNA (twice lower than 4%) selected out of 1,505 women examined at Kulakov National Medical Scientific Centre of Obstetrics, Gynaecology and Perinatal Medicine of the Ministry of Health of Russia during the last 2 years. 17 women were administered LMH and 32 women did not receive it. Group II included 113 pregnant women with a normal foetal DNA fraction (at least 4%), that were selected randomly out of patients who were examined during the same period. 56 pregnant women were administered LMH and 57 were not. Next Generation Sequencing was used as an NIPS method. In order to determine the foetal DNA level, we amplified DNA loci with single nucleotide polymorphisms. Study Results. In group I (low foetal DNA fraction), where no LMH therapy was initiated, median values were 3.5% (3.2–3.8%); if LMH was administered, these values made 3.6% (3.4–3.9), while in group II — 7.7% (6.1–9.5%) and 7.9% (6.0–10.5%), respectively; no significant differences (p = 0.29 and p = 0.7, respectively) were recorded in both cases. Conclusion. Use of LMH does not affect NIPS; therefore, it is not necessary to adjust the LMH therapy in pregnant women prior to NIPS. Keywords: non-invasive prenatal DNA screening, heparin, anticoagulants, low molecular heparins, habitual abortion, pregnancy complications.

Uterine leiomyomas are a worrying reproductive health issue that has serious social implications. The aim of this study was to conduct a search for marker single nucleotide polymorphisms (SNPs) associated with uterine leiomyoma. To test the hypothesis about the contribution of genetic predisposition to the pathogenesis of myomas, the initial group of 100 patients with a verified diagnosis of uterine leiomyoma was divided into 2 subgroups: subgroup Ia (women with a family history of the disease) and subgroup 1b (women with no family history of the disease). The control group consisted of 30 postmenopausal patients who did not have a medical history of uterine fibroids and denied uterine fibroids in their close female relatives. DNA sequences were read using Sanger sequencing. Statistically significant differences (p < 0.05) were discovered between the analyzed groups in terms of genotype frequencies for rs12637801 and rs12457644. Also, previously unknown protective SNPs were identified whose rare alleles could predict the reduced risk of uterine leiomyomas.

HPV infects epithelial tissues regardless of their location and penetrates into the cell through microscopic cuts. The clinical manifestations of HPV-associated diseases are diverse and equally relevant for women and men. Despite the fact that exophytic condylomas are regarded as a benign disease and establishing diagnosis does not cause difficulties, a clinician should always have oncological alertness, especially with regard to diffuse, long-lasting condylomatosis.

Relevance. Life expectancy of cystic fibrosis patients mostly depends on the degree of respiratory system damage caused by opportunistic microorganisms, which is due to the fact that 90-95% of deaths of cystic fibrosis patients are caused by lung infections. Goal. To define epidemiologic characteristics of chronic lung infection caused by the most common agents (S. aureus, P. aeruginosa, B. cepacia-like bacteria (Bcc) and Achromobacter spp.) using a novel chronic lung infection in cystic fibrosis patients microbiological diagnosis algorithm. Materials and methods. Over a period of 7 years (2008-2016) 300 children with cystic fibrosis living in Moscow, Moscow region and several other regions of Russian Federation have been checked-up. 260 sputum samples from 100 adult patients, who were under care at the Pulmonology Research Institute, were studied. Sputum samples from children were taken before and after antibiotic therapy with intervals of 15-45 days and over 6 months. 30 of the children were also subjected to a microbiologic monitoring of the state of chronic infection in the period between 4 and 15 months. Sputum sample from adult patients were also taken before and after antibiotic therapy with intervals of 0, 15-45 days and over 6 months. Results. P. aeruginosa, S. aureus, H. influenzae and Burkholderia cepacia-like bacteria were confirmed to be the most common agents of lung infection in cystic fibrosis patients. Children with cystic fibrosis over the years develop foci of chronic lung infection, mainly caused by P. aeruginosa and S. aureus. Conclusions. Chronic lung infection can be caused by community-acquired or nosocomial S. aureus и P. aeruginosa. Chronic lung infection is a complex, dynamically changing disease which requires constant monitoring and is mainly caused by S. aureus, P. aeruginosa, Bcc bacteria and Achromobacter spp. As populations of the agents can be diverse, it is necessary to study all colonies with differing phenotypes (mucoid and non-mucoid variants, small colony variants, variants with different pigments) and to take samples of several colonies when testing antibiotic resistance. Bcc and Achromobacter spp. cannot be eradicated with antibiotics, thus the only effective measure against these bacteria can only be vaccination which requires developing a vaccine.

A new method for selection of bacterium antibiotic resistance genes is proposed and tested for solving the problems related to selection of primers for PCR assay. The method implies clustering of similar nucleotide sequences and selection of group primers for all genes of each cluster. Clustering of resistance genes for six groups of antibiotics (aminoglycosides, β-lactams, fluoroquinolones, glycopeptides, macrolides and lincosamides, and fusidic acid) was performed. The method was tested for 81 strains of bacteria of different genera isolated from patients (K. pneumoniae, Staphylococcus spp., S. agalactiae, E. faecalis, E. coli, and G. vaginalis). The results obtained by us are comparable to those in the selection of individual genes; this allows reducing the number of primers necessary for maximum coverage of the known antibiotic resistance genes during PCR analysis.

The aim of this study was to determine the changes of metabolomic profiles in embryonic culture media (ECM) for the evaluation of quality and implantation potential of human embryos. ECM (n=163) were collected on day 5 before transfer or cryopreservation. Some embryos were used in preimplantation genetic screening for detection of aneuploidy karyotypes. Samples were subdivided into groups according to embryo morphological classification (by Gardner), genetic analysis and implantation data. ECM were extracted with methanol, precipitates were separated by centrifugation and metabolite production of individual embryo was analysed by LC-MS (the positive ion mode). After peak detection and retention time alignment, data were analysed using the PCA algorithm. MS fingerprinting analysis of embryo culture medium showed significant differences between morphologically divided groups. Intragroup comparisons did not reveal differences between subclasses. Genetic screening of embryos revealed 33 aneuploid karyotypes. It was shown that chromosome number did not affect the metabolite profiles comparing with the normal group. The culture media of embryos that were positive or negative for successful implantation showed specific signatures that allowed to distinguish embryos with different outcomes.The characterization of ECMs by LC-MS may facilitate more accurate selection of the best embryo for the implantation, improving single-embryo transfer and thus eliminating the risk and undesirable effects of multiple pregnancies.Tsel'iu issledovaniia iavlialos' opredelenie izmeneniia profileĭ metabolitov v émbrional'nykh pitatel'nykh sredakh (ÉPS) dlia otsenki kachestva i implantatsionnogo potentsiala kul'tiviruemykh émbrionov cheloveka. ÉPS (163 sht.) byli sobrany na 5-ĭ den' do perenosa ili kriokonservatsii émbrionov. Chast' émbrionov voshla v programmu predimplantatsionnogo geneticheskogo skrininga s tsel'iu obnaruzheniia aneuploidnykh kariotipov. Obraztsy byli razdeleny na gruppy soglasno morfologicheskoĭ klassifikatsii émbrionov (po Gardneru) v sootvetstvii s dannymi geneticheskogo analiza i informatsii ob implantatsii posle perenosa. ÉPS ékstragirovali metanolom, osadok otdeliali tsentrifugirovaniem i produktsiiu metabolitov otdel'nykh émbrionov detektirovali s ispol'zovaniem VÉZhKh-MS v rezhime polozhitel'nykh ionov. Posle obnaruzheniia pikov i vyravnivaniia khromatogramm dannye obrabatyvali pri pomoshchi analiza glavnykh komponent (PCA). Profilirovanie metabolitov v ÉPS vyiavilo sushchestvennye razlichiia mezhdu morfologicheskimi klassami émbrionov. Vnutrigruppovye sravneniia ne obnaruzhili dostovernykh otlichiĭ mezhdu predstaviteliami odnogo klassa. Geneticheskiĭ skrining émbrionov vyiavil 33 anéuploidnykh kariotipa. Bylo pokazano, chto chislo khromosom ne vliiaet na profil' metabolitov po sravneniiu s normal'noĭ gruppoĭ émbrionov togo zhe klassa. V ÉPS implantirovavshikhsia i neimplantirovavshikhsia émbrionov byli vyiavleny spetsificheskie molekuliarnye signatury, pozvoliaiushchie differentsirovat' émbriony s raznym iskhodom implantatsii. Predpolagaetsia, chto profilirovanie metabolitov v ÉPS s pomoshch'iu VÉZhKh-MS mozhet sposobstvovat' bolee tochnomu otboru dlia perenosa odnogo émbriona s tsel'iu snizheniia riska nezhelatel'nykh posledstviĭ mnogoplodnoĭ beremennosti.

Objective: to investigate the distribution of the genotypes and alleles of the PTPN22, TNFAIP3, CTLA4, TNFA, IL6, IL6R, IL10, MCP1, and ICAM1 genes in patients with rheumatoid arthritis (RA) and in the control group of healthy individuals, to estimate their significance as molecular genetic markers for predisposition to RA; and to analyze the correlation between the gene polymorphisms included in the study and the production of anti-cyclic citrullinated peptide antibodies (ACCPA) and IgM rheumatoid factor (RF).Subjects and methods. The investigation was conducted within the framework of the «Early arthritis: Diagnosis, outcome, criteria, active treatment program». The prospective follow-up study included 122 patients with RA fulfilling the 1987 American College of Rheumatology (ACR) criteria; with disease duration of ≤ 2 years. 73 (59.8%) patients were included during the first 6 months after the onset of the disease. 74 (60.7%) and 81 (66.5%) patients were found to be positive for ACCPA and IgM RF, respectively. 314 healthy blood donors served as a control group. A real-time polymerase chain reaction was used in the patients and control individuals to study the distribution of the polymorphic variants of PTPN22 (+1858 C >T, rs2476601), TNFAIP3 (rs675520, rs6920220, rs10499194), CTLA4 (+49A>G, rs231775 ), TNFА (-308A>G, rs1800629), IL6 (-174G>C, rs1800795), IL6R (+358A>C, rs8192284), IL10 (-592A>C, rs1800872, -1082 A>G, rs1800896), MCP1/CCL2 (+2518A>G, rs1024611), and ICAM1 (721G>A, rs1799969) genes. Results and discussion. This analysis revealed an association of PTPN22 (+1858 C >T, rs2476601) and TNFAIP3 (rs675520, rs10499194) polymorphisms with the risk of RA (odds ratio (OR), 1.5; 95% confidence interval (CI), 1.0–2.3; p = 0.05; OR, 1.5; 95% CI, 1.1–2.0; p = 0.02; OR, 0.5; 95% CI, 0.4–0.8; p = 0.01, respectively. Further, there was a tendency towards a positive association of TNFAIP3 (rs6920220) and IL6R (rs8192284) polymorphisms with a predisposition to RA (p = 0.056). IL6 (rs1800795), IL10 (rs1800872, rs1800896), MCP1/CCL2 (rs1024611), and ICAM1 (rs1799969) polymorphisms were not associated with the risk of RA. An analysis of the findings after patient stratification by ACCPA and IgM RF (a binary variable) showed that none of the polymorphisms in question was associated with RF state. At the same time, PTPN22 (rs2476601), TNFAIP3 (rs675520), TNFAIP3 (rs10499194), and TNFА (rs1800629) polymorphisms were found to be significantly related to ACCPA state (a binary variable). The level of ACCPA as a quantitative variable was statistically significantly associated with CTLA4 (rs231775) and TNFА (rs1800629) polymorphisms in a dose-dependent fashion (р = 0.025 and р = 0.015, respectively). There was a marked tendency towards an association of ACCPA levels and IL6R gene polymorphism (p = 0.07). IL6 (rs1800795), IL10 (rs1800872, rs1800896), MCP1/CCL2 (rs1024611), and ICAM1 (rs1799969) polymorphisms were not correlated with ACCPA state (binary and quantitative variables).Conclusion. The findings suggest that a number of genes are implicated in the pathogenesis of RA and that they are involved in the development of ACCPA-positive and ACCPA-negative RA subtypes. No relationship was found between the production of IgM RF and the polymorphisms of the genes under study. The findings suggest that there appears to be different mechanisms for the formation of autoantibodies (ACCPA and IgM RF) in RA.

Aim. To identify gene polymorphisms (OPG, RANKL, VDR, SOST) and evaluate their relationship with bone mineral density (BMD) and fractures for further optimization of diagnosis and treatment of postmenopausal osteoporosis (PMO). Materials and methods. The study included 236 postmenopausal women living in Moscow and the Moscow region. The main group (I) consisted of 174 patients, aged 50 to 83 years, with BMD < - 2.5 SD for the T-score. The comparison group (II) consisted of 62 postmenopausal women with BMD within the normal range and the lack of previous fractures, aged 50 to 77 years. Molecular genetic analysis of polymorphisms of VDR (rs10735810, rs1544410), RANKL (rs9594738, rs9594759) and OPG (rs3102735, rs3102735 and rs4355801), SOST (rs1230399) was performed by polymerase chain reaction in all subjects with PMO and in the control group. Results. In the presence of the T allele of the gene RANKL (rs9594759 and rs9594738) risk of reduced BMD at L1-L4 was increased by 2 times. Women with genotype C/C gene polymorphism of OPG (rs3102735) the risk of fractures of the distal radius (PR) was increased by 17 times, regardless of BMD. Genotype G/G rs1544410 polymorphism of VDR gene in patients with PMO is associated with an increased risk of fractures of the distal radius by three times. In patients with PMO genotype C/C in gene SOST (rs1230399) only cause differences in body mass index. According to an autosomal recessive pattern, homozygous genotype C/C was significantly associated with obesity in the PMO. Conclusions. Polymorphisms of genes RANKL (rs9594759 and rs9594738), OPG (rs3102735) and VDR (rs1544410) are associated with the risk of fractures and PMO. Gene SOST (rs1230399) had no statistically significant effect on BMD and fracture risk.

The review examined the role of HPV in anogenital diseases. There are presented modern data on the prevalence of anal intraepithelial neoplasia (AIN), HPV infection of the anal region among women at risk (cervical intraepithelial neoplasia - CIN, vulvar - VIN, vaginal - VaIN). There are considered methods of diagnosis of HPV-associated anogenital diseases. Analyzes the literature on the importance of the development of screening AIN in patientswith CIN, VIN, VaIN. There are presented data of the preliminary results of the research of HPV infection of the anal region in patients with genital neo-plasias, the data on different tropism of HPV to cervical epithelium and the epithelium of the anal region.

Background. The early diagnosis of ovarian cancer (OC) is an important problem in modern gynecological oncology due to significant detection rates for late-stage tumors. Intensive screening of patients from high-risk groups that include OC predisposition gene mutation carriers is indicated.Subjects and methods. An unselected group of 202 patients with OC and two control groups of blood donors: 591 healthy females; 1197 persons (including 591 females, 606 males) were examined. Patients and healthy individuals who identified themselves as ethnic Russians and residents of the Russian Federation participated in the study. Whole peripheral blood samples were collected at the Clinical Subdivisions of the N.N. Blokhin Russian Cancer Research Center and at the Department of Transfusiology of the Acad. B.V. Petrovsky Russian Research Center of Surgery in 2012–2013. Informed consent was obtained from all the participants. DNA was extracted using a Prep-GS-Genetics reagent kit. Real-time polymerase chain reaction genotyping assay was carried out by melting-curve analysis employing an BRCA SNP genotyping kit(BRCA1 and BRCA2 gene mutations) and original oligonucleotides (CHEK2, NBN, and BLM gene mutations). Thirteen population-specific mutations, including 7 (185delAG, 4153delA, 5382insC, 3819delGTAAA, 3875delGTCT, 300T>G, and 2080delA) in the BRCA1 gene, 1 (6174delT) in the BRCA2 gene, 3 (1100delC, IVS2+1G>A, and 470T>C) in the CHEK2 gene, 1 (657delACAAA) in the NBN gene, and 1 (1642C>T) in the BLM gene, were genotyped. Polymerase chain reaction was performed using a DTprime real-time detection thermal cycler.Results and discussion. BRCA1 and BRCA2 gene mutations were detected in 46 (22.8 %) patients with OC; the prevailing mutation in the BRCA1 gene was 5382insC (58.7 %). OC was diagnosed in 32.6 % of the patients aged 51 years or older. The rate of moderate-penetrance mutations (1100delC and IVS2+1G>A in the CHEK2 gene, 657del5 in the NBN gene, and 1642C>T in the BLM gene) was 0.5–1.0 % in the group of OC patients and 0–0.3 % in the control group of healthy women. The majority of these patients (5/6) were diagnosed with OC at age less than 50 years. The CHEK2 mutation, 470T>C, was more frequently encountered in the control group (6.6 %) than in the OC patient group (5.0 %). High rate of the CHEK2 mutation, IVS2+1G>A, was first shown for OC patients in the Russian Federation, the odds ratio was 11.9 (95 % confidence interval, 9.5–14.3; p = 0.056). It was preliminarily concluded that it played an important role in the development of OC in the Russian population; our findings should be verified in further investigations. The difference in the rate of mutations, such as 1100delC, IVS2+1G>A and 470T>C in the CHEK2 gene, 657del5 in the NBN gene, 1642C>T in the BLM gene, were insignificant in the patient and control groups, which may be related to the low population rate of these genetic markers and, in case of the CHEK2 mutation, 470T>C, may be linked to its low penetrance. By taking into account the fact that numerous studies have proven the clinical significance of all examined moderate-penetrance mutations whose prevalence in the Russian population has been confirmed by the authors of this paper, the inclusion of the mutations in a diagnostic panel to detect hereditary predisposition to OC is substantiated. The associated risks are higher for the rare mutations leading to the formation of truncated nonfunctional proteins, which are 1100delC and IVS2+1G>A in the CHEK2 gene, 657del5 in the NBN gene, and 1642C>T in the BLM gene. The penetrance of the CHEK2 mutation, 470T>C, is lower, which should be kept in mind during medical genetic counselling.Conclusion. The total rate of mutations in the BRCA1, BRCA2, CHEK2, NBN, and BLM genes in patients with OC was 30.7 %, which determines the expediency of molecular genetic screening in this category of patients.

Golovatyuk K. P., Nosenko O. M., Makshayeva E. T., Trofimov D. Yu., Donnikov A. E., Kolin V. V. Molecular-genetic aspects of the endometrium state on the day of the tentative implantation window in women with recurrent miscarriage in the programs of assisted reproductive technologies. Journal of Education, Health and Sport. 2017;7(9):621-626. eISSN 2391-8306. DOI http://dx.doi.org/10.5281/zenodo.1068361 http://ojs.ukw.edu.pl/index.php/johs/article/view/5076 The journal has had 7 points in Ministry of Science and Higher Education parametric evaluation. Part B item 1223 (26.01.2017). 1223 Journal of Education, Health and Sport eISSN 2391-8306 7 © The Authors 2017; This article is published with open access at Licensee Open Journal Systems of Kazimierz Wielki University in Bydgoszcz, Poland Open Access. This article is distributed under the terms of the Creative Commons Attribution Noncommercial License which permits any noncommercial use, distribution, and reproduction in any medium, provided the original author(s) and source are credited. This is an open access article licensed under the terms of the Creative Commons Attribution Non Commercial License (http://creativecommons.org/licenses/by-nc/4.0/) which permits unrestricted, non commercial use, distribution and reproduction in any medium, provided the work is properly cited. This is an open access article licensed under the terms of the Creative Commons Attribution Non Commercial License (http://creativecommons.org/licenses/by-nc/4.0/) which permits unrestricted, non commercial use, distribution and reproduction in any medium, provided the work is properly cited. The authors declare that there is no conflict of interests regarding the publication of this paper. Received: 15.08.2017. Revised: 20.09.2017. Accepted: 29.09.2017. MOLECULAR-GENETIC ASPECTS OF THE ENDOMETRIUM STATE ON THE DAY OF THE tentative IMPLANTATION WINDOW INWOMEN WITH recurrent miscarriage IN THE PROGRAMS OF ASSISTED REPRODUCTIVE TECHNOLOGIES 1K. P. Golovatyuk, 2O. M. Nosenko, 1E. T. Makshayeva, 3D. Yu. Trofimov, 4A. E. Donnikov, 3V. V. Kolin 1Limited Liability Company Medical Center for Reproductive Health "Gameta", Odessa, Ukraine; 2Odessa National Medical University, Ukraine; 3Limited Liability Company "NPF DNA-Technology", Moscow, Russia; 4FGBU Scientific Center for Obstetrics, Gynecology and Perinatology. named after acad. V.I. Kulakov Ministry of Health care and Social Development of Russia, Moscow, Russia; e-mail: nosenko.olena@gmail.com Abstract More than 50% of pregnant women after the programs of assisted reproductive technologies (ART) face the problem of recurrent miscarriage (RMC), especially in the first trimester. Significant role in the development of RMC has infectious factor and chronic inflammation in the endometrium. The aim: to reveal the peculiarities of immune response mRNA genes of the inflammatory component expression in the period of the tentative implantation window (TIW) in women with RMC in ART programs. Material and methods. The main group consisted of 240 patients with RMC in ART programs; the control group included 100 conditionally healthy fertile women. On the ground of PCR reverse transcription, the mRNA of the IL-1β, IL-2, IL-10, Foxp3, TLR9, IL-2Rα cytokine genes was examined in endometrial samples obtained with the help of biopsy on the TIW day. Results. Analysis of the transcriptional profile of the immune response genes in the endometrium on TIW day revealed that the relative level of mRNA expression of the IL-1β, IL-2, Foxp3, TLR9, IL-2Rα genes did not differ significantly in the main and control groups. Statistically significant decrease in mRNA expression of IL-10 gene was observed in women with RPL. Conclusions. A feature of mRNA expression of the inflammatory component of the immune response in TIW period in women with RMC in ART programs is a decrease in the expression level of the IL-10 gene mRNA, which may be one of the reasons for the unfavorable outcomes of the onset pregnancy. Key words: recurrent miscarriage, assisted reproductive technologies, immune response, reverse transcription-polymerase chain reaction, tentative implantation window.

A highly sensitive test system, based on the immuno-PCR method, was developed for the detection of two staphylococcal toxins: enterotoxin A (SEA) and toxic shock syndrome toxin (TSST). A key element of the developed systems was to obtain supramolecular complexes of bisbiotinylated oligodeoxynucleotides and streptavidin, which were to be used as DNA-tags. Specificity studies showed no cross-reactivity when determining SEA and TSST. The sensitivity of detection of these toxins in the culture supernatants S. aureus was not lower than 10 pg/mL.

The viral nature of many female genital cancers is now beyond question. By taking into account this fact, the problem of qualitative assessment ofthenatureofcervicalintraepithelialneoplasia(CIN)anditsfocusonprogressiontoinvasivecarcinomabecomesquitenatu ral.Studiesof a number of biological markers of carcinogenesis have recently provided a possibility for prospective prediction. The paper con siders the as- pects of importance of the molecular biological markers of proliferation and apoptosis in the etiopathogenesis of genital cancers. It gives the results of examinations of 16 patients with histologically verified squamous cell carcinoma of the cervix uteri, 40 patients di agnosed as having CIN of different grades (CIN-1, CIN-2, CIN-3 — squamous cell carcinoma in situ), and 6 patients with the morphologically unaltered cervi- calepithelium,whosecervicalscrapeswereanalyzedfortheexpressionofthemRNAgenesССNB1,Ki-67,BA G,BCL-2,ESR1,andPRG. It is shown that the molecular genetic findings may be new prognostic markers that reflect the possible disease developmental pathways, sug- gesting the need for further investigation of biomarkers in order to prevent malignancies and to reduce their morbidity.

We propose a method of quantitative functional activity assessment in cells isolated via sorting on a flow cytometer. We show that cell populations vary in the mRNA expression of cytokine genes immediately after isolation and sorting, while the maintenance of homogenous populations in culture without stimulation results in an increase in these gene mRNA expression. Using the original system, it is now possible to detect mRNA cytokine genes with high sensitivity, starting from 90 cells per specimen. This approach permits genetic and immunogenetic analysis of gene expression with the goal of determining their functions in the in vitro studies.

Background.. To investigate expression of cytokines genes parameters in skin and blood in severe atopic dermatitis (AD) and cutaneous T-cell lymphoma (CTCL) patients comparing with healthy donors. Materials and methods. 20 severe AD patients, 20 CTCL patients and 20 healthy donors were included in the study. Skin samples and peripheral blood were used as material for immunological study. Interleukins — (IL)1B, IL2, IL2r, IL4, IL5, IL6, IL7, IL8, IL10, IL12A, IL12B, IL15 (total), IL15 , IL17A, IL18, IL23, IL28, IL29, Interferon γ (IFNγ), tumor necrosis factor α (TNFα), transforming growth factor beta 1 (TGFB1), forkhead box P3 (FOXP3) gene expression was defined in the skin and peripheral blood of severe AD patients, CTCL patients and healthy donors by real-time reverse transcription polymerase chain reaction (RT-PCR). Results. Statistically significant increase of cytokines genes IL4,IL12B,IL17A, TNFα in peripheral blood of severe AD patients compared with CTCL patients was marked. Studying of skin samples from CTCL patients has shown statistically significant increase of cytokines IL8, IL10, IL15, IFNγ genes expression and decrease of IL18 gene expression in comparison with skin samples from severe AD patients. Conclusion. Obtained cytokines genes expression cytokines genes parameters in peripheral blood of severe AD and CTCL patients had a certain similarities consisting of increased IL8, IFNγ and decreased IL6, IL23 genes expression in comparison with healthy donors. Substantial differences in peripheral blood of patients in comparison with healthy donors: increased IL-5, IL-12B genes expression in AD patients and decreased IL-8, IL-12A genes expression in CTCL patients, at the same time increased expression of IL-6, IL-8, IL-10, IFNγ genes in severe AD and CTCL patients were shown.

Цель: Изучить эффективность золедроновой кислоты в терапии постменопаузального остеопороза (ПМО) с учетом полиморфизма гена фарнезилдифосфат-синтетазы (FDPS). Методы исследования: клинический, определение биохимических маркеров костного метаболизма (БМКР) - электрохемилюминесцентным методом («Хоффманн-Ла Рош ЛТД», Швейцария). Измерение МПК проводили с помощью двухэнергетической рентгеновской абсорбциометрии позвонков поясничной области (L1-L4) и шейки бедренной кости (Neck left, Neck total left). Молекулярно-генетический (определение однонуклеотидного полиморфизма гена FDPS (rs2297480) - методом «примыкающих проб» с анализом кривых плавления (ЗАО «НПФ ДНК-Технология, Россия)). Результаты. Обследовано 225 женщин с постменопаузальным остеопорозом. Средний возраст составил 59 (5466) лет. Средняя продолжительность периода постменопаузы 7 (2-13) лет. ИМТ=27,2(23,6-29,05) кг/м2. Распределение аллелей (АА:АС:СС) составило 55,1(n=144):39,5(n=103):5, 4(n=14). Базальные уровни биохимических маркеров костного ремоделирования (БМКР) соответствовали периоду постменопаузы и статистически значимо не различались в зависимости от генотипа FDPS. Средние показатели МПК в поясничном отделе позвоночника и/или шейке бедра соответствовали остеопорозу. Всем пациенткам проводили терапию золедроновой кислотой (zol) в дозе 5 мг в виде внутривенной инфузии 1 раз в 12 месяцев в течение 2 лет. Дополнительно все пациенты получали кальций 1000 мг/сут и витамин Д3 800МЕ/сут. Реакция БМКР на инфузию zol различалась в зависимости от генотипа FDPS. Через 6 месяцев после 1-ой инфузии достоверно более выраженное снижение B-Crosslaps 92 (91-93)% отмечено у пациентов с С/С генотипом (p=0,056), по сравнению с 82% у носителей аллеля А (p=0.07). Через 9 месяцев после инфузии B-Crosslaps у носителей аллеля А повышались до 75%, остеокальцина - 44,4% от исходного. Тогда как у носителей генотипа C/C как B-Crosslaps - (87%), так и osteocalcine -(53,6%) оставались по-прежнему низкими (р=0,056). К 12 месяцу после инфузии уровни B-Crosslaps и osteocalcin у носителей аллеля A повышались и соответствовали премено-паузальным значениям, тогда как у носителей С/С генотипа оставались в пределах ниже нижней границы нормы для пременопаузы. У пациентов с СС генотипом на фоне выраженного снижения БМКР после 2ой инфузии золедроновой кислоты произошло снижение МПК на 5,6%. Выводы. Различная реакция на терапию золедроновой кислотой у пациенток с ПМО связана с полиморфизмом гена FDPS (rs2297480). Для пациентов с генотипом С/С характерно «гиперторможение» костного обмена в ответ на терапию золедроновой кислотой, что возможно, было причиной отрицательной динамики МПК в поясничном отделе позвоночника на втором году терапии золедроновой кислотой.

Aneuploidies as quantitative chromosome abnormalities are a main cause of failed development of morphologically normal embryos, implantation failures, and early reproductive losses. Preimplantation genetic screening (PGS) allows a preselection of embryos with a normal karyotype, thus increasing the implantation rate and reducing the frequency of early pregnancy loss after IVF. Modern PGS technologies are based on a genome-wide analysis of the embryo. The first pilot study in Russia was performed to assess the possibility of using semiconductor new-generation sequencing (NGS) as a PGS method. NGS data were collected for 38 biopsied embryos and compared with the data from array comparative genomic hybridization (array-CGH). The concordance between the NGS and array-CGH data was 94.8%. Two samples showed the karyotype 47,XXY by array-CGH and a normal karyotype by NGS. The discrepancies may be explained by loss of efficiency of array-CGH amplicon labeling.

An increasing number of hormonal contraceptives and their varying system effects on the body of a woman makes it necessary to personify their prescription and search for means of predicting their safety and tolerability. The article presents the research data and shows the various informative clinical predictors in the development of adverse reactions in the background. It was found that the most significant clinical predictors of occurrence of side effects and complications on the background of the use of the HC are the existence of gynecological diseases associated with menstrual cycles and poor portability of HC in history; an independent predictor of adverse effects on the background of the COC are chronic cholecystitis and dysfunction of the sphincter of Oddi; dynamics of lipid parameters, biochemical blood spectrum, as well as some parameters of hemostasis with the use of different variants of HC; genotype women polymorphic locus rs2414096 aromatase gene (CYP19A1) is a significant predictor of complications and side effects when using HC.

The PCR method in the real-time mode helped to analyze the quantitative characteristics of the normal and conditionally pathogenic aerobic/optionally anaerobic, anaerobic biota, mycoplasmas and Candida fungi in the urethra, cervical channel and vagina in healthy women aged 18-45. The analysis revealed absence of any differences between the biotopes by the bacterial spectrum and quantity of both aerobic and anaerobic bacteria with regard to Lactobacillus spp. The biota of all of the women who were younger than 40 and some of the women who were older than 40 had mostly lactobacilli. The microbial composition of the biocenosis in some women older than 40 is characterized by the reduction in the quantity of lactobacilli and their replacement with anaerobic microorganisms/groups of microorganisms, mainly such as Atopobium vaginae, Megasphaera spp./Veillonella spp./Dialister spp., Gardnerella vaginalis/Prevotella bivia/Porphyromonas spp. and Eubacterium spp. Any change in the microbial composition of the urogenital tract biota can serve as a marker of hypoestrogenia.

The virus particles of live mumps virus vaccine widely used for vaccination in Russia have been detected and visualized by the atomic force microscopy. For quantitative estimation of the number of observed virus particles the special method has been developed. The presence of the vaccine virus protein component was tested by ELISA and dot-blot analysis. Using a quantitative real-time PCR assay the number of copies of viral RNA was estimated. The results of the quantitative estimation obtained by real-time PCR corresponded to the atomic force microscopy data.

Glanders and melioidosis are serious infectious diseases of humans and animals. The pathogens of these infections are potential group B bioterrorism agents. In this article we studied the possibility of using primers based on the flagellar gene to identify Burkholderia mallei and Burkholderia pseudomallei and to diagnose experimental glanders and melioidosis. On the basis of the results it was concluded that PCR using the primers developed can be recommended for inclusion in the scheme of laboratory diagnosis of glanders and melioidosis, both for the identification of pure cultures and in the investigation of experimental clinical material.

We studied the relationship between the levels of protamines 1 and 2 (PRM1 and PRM2) and fertilin-β (ADAM-2) mRNA expression and outcomes of infertility treatment using assisted reproductive technologies was studied. Analysis of the relationships between the outcomes of in vitro fertilization and embryo transfer and profiles of the expression of seminal genes PRM1, PRM2, ADAM-2 mRNA, evaluated by reverse transcription quantitative PCR was carried out in 79 couples. Significant differences in the expression of seminal PRM1, PRM2, ADAM-2 mRNA were detected in couples with different outcomes of in vitro fertilization and embryo transfer. The levels of seminal gene expression are potential predictors of the efficiency of in vitro fertilization and embryo transfer.

The expression of mRNA of cytokines and immunoregulatory molecules characterizing the interaction of mesenchymal stromal cells from chorionic villi of postpartum placenta and allogenic mononuclear blood cells was studied during 3-day co-culturing of these cells. The expression of foxp3, il2ra, and il10 mRNA in floating mononuclear cells increased from day 1 to 3 in co-culture, which can refl ect the process of induction of regulatory T cells in the lymphocyte population under the action of mesenchymal stromal cells.

Studies of collagen gene polymorphisms associated with predisposition to early recurrent miscarriages revealed significant differences in the distribution of COL1A1 C-1997A C>A (rs1107946) genotypes and alleles in the group of pregnant patients with early miscarriages in comparison with controls (normal pregnancy). Identification of COL1A1 C-1997A C>A (rs1107946) collagen gene polymorphisms at the stage of pregnancy planning will make it possible to form early miscarriage risk groups for more thorough preparation to gestation and optimization of follow up of this patient population.

mRNA of genes (tgfβ, il6, il10, il1β, and vegf165) involved in cell proliferation, inflammatory, anti-inflammatory, and angiogenic processes were detected and quantified in cultures of mesenchymal stromal cells of different passages derived from human extraembryonic tissues (amniotic sac, umbilical cord, chorionic villi and trophoblast of the placenta). The concentrations of IL-10, IL-6, IL-1β, and TGFβ were measured.

The accuracy of a real-time polymerase-chain-reaction assay for genetically modified sources in food products was determined using two official test systems (kits) of primers and samples. These kits were recommended by the Federal Center of State Sanitary and Epidemiological Surveillance (Russian Ministry of Health) and the European Commission. We used the following three models of thermocyclers: iCycler iQ (Bio-Rad, United States), Rotor-Gene 3000 (Corbett Research, Australia), and DT-322 (DNA-Technology, Russia). Studies of samples that contained 1% genetically modified sources showed that the error of a quantitative assay for genetically modified sources in food products corresponds to 20–30% and does not depend on the kit type and the thermocycler model used.

To determine the correlation between interleukin 28B (IL28B) gene polymorphism in patients with chronic hepatitis C (CHC), the presence or absence a rapid virologic response to antiviral therapy, and a number of immunological characteristics as a basis for a personalized approach to treating the patients.Seventeen CHC patients infected with hepatitis C virus genotype 1b were examined and underwent genetic testing for IL28B gene polymorphism for rs12979860 (CC, CT or TT genotypes) and rs8099917 (TT, TG or GG genotypes) using the modified method of adjacent samples, which revealed single nucleotide substitutions in the genes. Their immunological parameters were identified by a flow cytometry technique by taking into account whether a rapid virologic response had been achieved.The key phenomena of a rapid virologic response in the representatives of different IL28B genotypes are the nonspecific proliferative activity of blood natural killer cells before treatment, as well as the count of regulatory T cells before and 4 weeks after therapy start.To predict the efficiency of antiviral therapy for CHC, it is desirable to supplement genetic studies with immunological data.

Population studies aimed at identifying genetic markers of diseases and at improving the selection of histocompatible donors for allografting suggest that basic research and applied studies of the system of Human Leukocyte Antigens (HLA) are best performed at the molecular-genetic level (DNA typing).