[Protocol of Transcriptome Analysis of Decidual Placenta Cells].

The advent of high-throughput sequencing technologies has expanded our understanding of the biological significance of non-coding regions of the genome. In recent years, more and more studies have been devoted to studying the role of noncoding RNAs in the development of diseases, as well as their participation in various cellular processes. Until now, all transcriptome studies of native placental tissue with the description of the noncoding RNA region were carried out without isolating individual cell populations. This approach, due to the high cellular heterogeneity of the placental tissue, significantly complicates the ability to determine the molecular-biological functions of individual cells and their role in the molecular pathogenesis of reproductive disorders. In this work, we propose a technique for obtaining total RNA from single decidual cells of frozen placental tissue obtained by laser-capture microdissection technology for transcriptome sequencing, including a cluster of noncoding RNAs. This technique can be successfully used to study the full-genome expression profile of other placental cell populations. The high accuracy of results on the transcriptome profiling of decidual cells obtained using the developed technique was additionally confirmed by an integrative analysis with the results of a 10x Genomics experiment.

[Natural Selection as a Driver for the Genetic Component of Preeclampsia].

Preeclampsia (PE) is a severe hypertensive pathology and affects 2-8% of pregnancies worldwide. Its etiopathogenesis is poorly understood, and prognostic biomarkers and effective treatments are unavailable for this pregnancy complication, determining the high rates of maternal and perinatal morbidity and mortality. Racial and ethnic differences in PE incidence are of interest to study in terms of evolutionary medicine because such variability can be considered as a side effect of adaptive changes that have occurred in the genetic structure of modern populations since the dispersal of Homo sapiens from Africa. Genetic diversity at 10 regulatory single nucleotide polymorphisms (rSNPs) associated with PE was studied in North Eurasian populations and world populations of the 1000 Genomes Project. The role of natural selection in the formation of this genetic diversity was assessed at the microevolutionary level. High interpopulation diversity was observed with the greatest contribution being made by allele frequencies of NDRG1 rs3802252 (FST = 0.157). Signatures of natural selection were detected for rs10423795 of LHB, rs2167270 of LEP, rs2227262 and rs3802252 of NDRG1, rs56153523 and rs8109071 of SYDE1, and rs72959687 of INHA. The results are consistent with two evolutionary hypotheses of PE, namely, those of ancestral susceptibility and genetic conflicts.

[The Role of Natural Selection in the Formation of the Genetic Structure of Populations by SNP Markers in Association with Body Mass Index and Obesity].

Obesity is one of the major challenges in modern society. More than a third of the world's population suffers froms overweight. This phenotype affects the quality of life and is associated with cardiovascular diseases, diabetes, cancer and reproductive disorders. The population variability of allele frequencies of 26 single nucleotide polymorphisms, in association with obesity and body mass index, according to data from genome-wide association studies (GWASs) is discussed in this study. Genetic variability was analyzed in populations of Northern Eurasia and populations from the human genome diversity project (HGDP). The population samples are characterized by high genetic diversity that correlates with climatic and geographical parameters. The results of the test for searching for natural selection signals revealed a selection effect for rs1167827 of the HIP1 gene, rs7138803 and rs7164727 located in the intergenic region, rs7141420 of the NRXN3 gene, rs7498665 of the SH2B1 gene, and rs7903146 of the TCF7L2 gene.

[Features of balloon angioplasty in the treatment of central vein stenosis in patients with arteriovenous fistula on programmed hemodialysis]. / Osobennosti primeneniya ballonnoi angioplastiki pri lechenii stenozov tsentral'nykh ven u bol'nykh s arteriovenoznoi fistuloi, poluchayushchikh lechenie programmnym gemodializom.

OBJECTIVE: To analyze AVF patency rates after balloon angioplasty and palliative open surgery in patients with native AVF and central vein stenosis. MATERIAL AND METHODS: The study included 39 patients who underwent percutaneous balloon angioplasty (main group) and 41 patients who underwent palliative interventions (comparison group): thrombectomy, proximalization of arteriovenous anastomosis, AVF blood flow reduction. All patients were diagnosed with stenosis of subclavian vein, innominate vein, inferior vena cava or multiple lesions. RESULTS: Primary annual patency rate in the main group was 15.4% [95% CI 6.2; 28.3], in the comparison group - 0% (p=0.0011). Secondary annual patency rate was 66.7% [95% CI 49.6; 79.1] and 19.5% [95% CI 9.2; 32.7], respectively (p<0.0001). In the main group, a strong negative correlation between primary patency rate and stenosis-free period was revealed (r= -0.627 [95%CI -0.787; -0.388], p<0.0001). We did not find such correlation in the comparison group (r=0,049 [95% CI -0.262; 0.351], p=0.7599). Thus, balloon angioplasty is less effective for delayed stenosis. This feature should be considered in planning vascular approach. Different primary and secondary patency rates in the comparison group indicate that AVF blood flow reduction is an effective palliative treatment. CONCLUSION: Balloon angioplasty can significantly increase duration of AVF patency. Effectiveness of this procedure significantly depends on the period of manifestation of central vein stenosis. Balloon angioplasty cannot be considered a radical method in case of significant stenosis. This method only prolongs period for creation of new permanent contralateral vascular approach. Reduction of AVF blood flow can significantly reduce clinical severity of central vein stenosis and slightly extends the period of AVF patency.

The interaction effect of angiogenesis and endothelial dysfunction-related gene variants increases the susceptibility of recurrent pregnancy loss.

PURPOSE: The role of genetic polymorphisms in the pathogenesis of recurrent pregnancy loss (RPL) has been studied intensively. Complex diseases, including miscarriage, are believed to have a polygenic basis, and gene-gene interactions can play a significant role in the etiology of the disease. This study was conducted to investigate the association of gene-gene interactions with angiogenesis, endothelial dysfunction-related gene polymorphisms, and RPL. METHODS: A case-control study was conducted with 253 unrelated RPL patients with 2 or more spontaneous pregnancy losses and 339 healthy women with no history of pregnancy complications. Genotyping of single-nucleotide polymorphisms (SNPs) was performed using real-time polymerase chain reaction (real-time PCR), restriction fragment length polymorphism (RFLP), or allele-specific polymerase chain reaction methods. RESULTS: The genotypes 677TT of the MTHFR gene, 936TT, 936CT, and 634CC, 634GC of the VEGF gene, and allele 894T of the NOS3 gene were associated with a predisposition to RPL in the Russian population. A significant role of additive and epistatic effects in the gene-gene interactions of the SNPs of SERPINE-1, ACE, NOS3, MTHFR, and VEGF genes in RPL was demonstrated. CONCLUSIONS: The results showed that gene-gene interactions are important for RPL susceptibility. Additionally, analysis of the genotype combinations of several allelic variants provides more information on RPL risk than analysis of independent polymorphic markers.

[Difficulties in diagnosis of dislocated permanent dialysis catheter followed by perforation of right internal jugular and massive hemothorax]. / Trudnosti diagnostiki dislotsirovannogo permanentnogo dializnogo katetera, privedshei k massivnomu gemotoraksu pri perforatsii pravoi vnutrennei iaremnoi veny.

Great vein perforation followed by massive hemothorax occurred after implantation of permanent dialysis catheter. Diagnostic difficulties in this case were caused by placement of catheter's shadow in the projection of right atrium on the frontal X-ray scan. Injury of initial segment of superior vena cava was diagnosed during emergency surgery for ongoing bleeding.

Quadrupole mass filter acceptance in stability island created by double-frequency quadrupole excitation.

Increase in quadrupole mass filter resolution at separation in narrow band stability island (X-band) formed by biharmonic resonance excitation of ion oscillation is discussed. X-band and the normal working quadrupole mass filter modes are compared at theoretical resolution of 10,000 and different separation times. Transmission curves, acceptance ellipses parameters, and acceptance characteristics are obtained by numerical simulation. Transmission coefficients are approximately the same in both modes. Dependence of acceptance ellipses parameters on ion inlet phases has a complicated oscillating form in the X-band mode. Acceptance contours calculated for given transition levels have been compared. At low acceptance level, the combined acceptance in the X-band mode was found to be one order of magnitude higher than in the normal mode.

[Association study of genetic markers of schizophrenia and its cognitive endophenotypes].

A replicative analysis of associations of 15 SNPs located in the regions of 11 genes (TCF4, VRK2, NOTCH4, ZNF804A, AGBL1, RELN, ZFP64P1, KCNB2, CSMD1, CPVL, NRIP1) and three intergenic regions (SLCO6A1/LINCOO491, LOC105376248/LOC105376249, SPA17/NRGN) with schizophrenia was conducted in the Russian population of the Siberian region. These SNPs were previously identified in genome-wide association studies (GWAS) of schizophrenia and cognitive abnormalities. The present study confirmed associations of KCNB2 rs2247572, CSMD1 rs2616984, and intergenic rs12807809 located in SPA17/NRGN with schizophrenia. It was established that the frequency of the CSMD1 rs2616984 G/G genotype was higher in patients compared to the control group (OR = 1.73; CI: 1.14­2.62; р = 0.0337). The frequencies of the KCNB2 rs2247572 TT genotype (OR = 0.41; CI: 0.20­0.87; р = 0.0485) and intergenic rs12807809 CT genotype located in SPA17/NRGN (OR = 0.70; CI: 0.53­0.94; р = 0.0464) were significantly decreased in patients compared to the control group.

[Panel of X-linked single-nucleotide polymorphic markers for DNA identification (XSNPid) based on multiplex genotyping by multilocus PCR and MALDI-TOF mass spectrometry].

Human genetic markers linked with the X chromosome (X-linked) are used in the field of population and medical genetics, as well as for DNA identification of individuals in forensic science and forensic medicine. We proposed an XSNPid panel that consists of 66 unlinked single nucleotide X chromosome markers and developed a protocol for their multiplex genotyping using multilocus PCR and MALDI-TOF mass spectrometry. The XSNPid panel is genotyped within two multiplexes (36 and 30 markers). The developed protocol provides an efficient genotype reading; the fraction of determined genotypes is 98.29%. The high level of gene diversity (0.461) for the X-linked SNPs included in the panel is characteristic of the Russian population. A total of 63 out of 66 markers that provide a high efficiency of genotyping and independent inheritance are suitable for DNA identification purposes. The XSNPid panel is characterized by a very high discriminating ability when studying the Russian population. The probability of genotype coincidence in two unrelated individuals is 9 × 10^(-27) for women and 2 × 10^(-18) for men. Also, the XSNPid panel has a greater multiplex capacity in addition to a higher discriminating ability compared to the other closest analogues of the X chromosome SNP sets, which makes it more cost effective and less time consuming. The XSNPid panel is a convenient tool, not only for individual DNA identification, but also for population genetic studies.

[Detection of novel genetic markers of susceptibility to preeclampsia based on an analysis of the regulatory genes in the placental tissue].

Regulatory single nucleotide polymorphisms (rSNPs) are the least-studied group of SNP; however, they play an essential role in the development of human pathology by altering the level of candidate genes expression. In this work, we analyzed 29 rSNPs in 17 new candidate genes associated with preeclampsia (PE) according to the analysis of the transcriptome in placental tissue. Three ethnic groups have been studied (yakut, russian, and buryat). We have detected significant associations of PE with eight rSNPs in six differentially expressed genes, i.e., rs10423795 in the LHB gene; rs3771787 in the HK2 gene; rs72959687 in the INHA gene; rs12678229, rs2227262, and rs3802252 in the NDRG1 gene; rs34845949 in the SASH1 gene; and rs66707428 in the PPP1R12C gene. We used a new approach to detecting genetic markers of multifactorial diseases in the case of PE based on a combination of genomic, transcriptomic, and bioinformatic approaches. This approach proved its efficiency and may be applied to detecting new potential genetic markers in genes involved in disease pathogenesis, which reduces missing heritability in multifactorial diseases.

[Evolution of genetic diversity and human diseases].

The problem of development and dispersion of complex diseases in human populations requires new views, approaches, hypotheses, and paradigms. Evolutionary medicine provides one of the promising approaches to this problem, putting the disease into an evolutionary context. Unlike classic approaches oriented to proximate issues on structure and mechanisms of a disease, evolutionary considerations are broader. It provides the basis for understanding the origin, dispersion, and maintenance of the high frequencies of pathological phenotypes in modern human populations. In the current paper, we try to review the modern concepts on the evolution of human genetic diversity, to shape the outlines of evolutionary medicine, and to illustrate evolutionary medical problems using our experimental data. Data on genome-wide search for the signals of decanalization and adaptation in the human genome and on related biological processes and diseases are presented. Some hypotheses and concepts of evolutionary medicine may be productive for revealing the mechanisms of origin and dispersion of complex diseases and for pathogenetics of multifactorial diseases. One of such concepts is the hypothesis of decanalization of genome­phenome relationships under natural selection during modern human dispersion. Probably, the high frequency of alleles associated with complex diseases (and partially the high prevalence of diseases themselves) could be explained in the framework of the hypothesis.

[Haplotype Analysis of Oculopharyngeal Muscular Dystrophy (OPMD) Locus in Yakutia].

Oculopharyngeal muscular dystrophy (OPMD) is a hereditary neuromuscular disease with autosomal dominant and rarely with autosomal recessive inheritance types. This study included 50 patients with a clinical diagnosis of OPMD, 23 asymptomatic carriers of the mutation from 45 unrelated families, and 56 healthy relatives, as well as population samples of four ethnic groups of Yakutia: Yakuts, Evens, Evenks, Yukaghirs. It was found that the cause of OPMD development in all investigated families is the same increase in.GCN repeats to 14 copies in the PABPN1 gene. The molecular structure ofthe (GCN)14 mutant allele is (GCG)10(GCA)3GCG. The genetic variability of ten SNPs at the OPMD locus was studied in patient families and population samples. The haplotypes of OPMD were determined by a segregation analysis technique using the EM algorithm in the groups of patients, mutation carriers, and population samples. Only one haplotype of four SNPs (ATCG) linked with the (GCN)14 mutant allele was found in Yakuts and Russian patients and OPMD mutation carriers. Probably, this indicates the accumulation of mutations as a result of the founder effect.

[Genetic diversity of X-linked STR-markers in Siberian populations].

X-chromosome markers are informative tool for studying of the genetic diversity in human populations and become useful for DNA identification when certain complex kinship cases need to be unraveled. In this work, we present population genetic data of 10 X-chromosome STRs (DXS8378, DXS9898, DXS8377, HPRTB, GATA172D05, DXS7423, DXS6809, DXS7132, DXS101 and DXS6789). Average level of expected heterozygosity (He) in populations under study was 0.73. Total level of genetic differentiation for 10 studied populations was relatively low (Fst = 0.031 comparing to autosomal and Y-chromosomal markers. Allele frequencies and parameters of forensic interest for panel of X-STRs were calculated. The overall values of PDf and PDm are 0.9999999997, 0.999998 respectively. The overall level of genetic differentiation for 10 population (Fst) is 0.03186.

[Prevalence of gene polymorphisms associated with immune-dependent diseases in the populations of North Eurasia].

The data on distribution of genetic diversity in gene polymorphisms associated with autoimmune and allergic diseases and with regulation of immunoglobulin E and cytokines levels in 26 populations of the Northern Eurasia is presented. Substantial correlation between the values of average expected heterozygosity by 44 gene polymorphisms with climatic and geographical factors has not been revealed. Clustering of population groups in correspondence with their geographic locations is observed. The degree of gene differentiation among populations and the selective neutrality of gene polymorphisms have been assessed. The results of our work evidence the substantial genetic diversity and differentiation of human populations by studied genes.

[Analysis of Eight Polymorphic Alu Elements in the Teleuts Population].

Allele frequencies and genetic diversity in the population of Teleuts were assessed by the Alu repeat polymorphism at eight autosomal loci (ACE, APOA1, PLAT, F13, PV92, A25, CD4, Dl). For comparison, the study included previously obtained data on the Alu polymorphism in 19 indigenous populations of Siberia. On the dendrogram of genetic distances, the Teleut population is located in the cluster of Siberian ethnic groups, which are similar in origin, geography, and cultural traditions.

[Genetic Variability and Structure of SNP Haplotypes in the DMPK Gene in Yakuts and Other Ethnic Groups of Northern Eurasia in Relation to Myotonic Dystrophy].

The genetic variability of the DMPK locus has been studied in relation to six SNP markers (rs2070736, rs572634, rs1799894, rs527221, rs915915, and rs10415988) in Yakuts with myotonic dystrophy (MD) in the Yakut population and in populations of northern Eurasia. Significant differences were observed in the allele frequencies between patients and a population sample of Yakuts for three SNP loci (rs915915, rs1799894, and rs10415988) associated with a high chance of disease manifestation. The odds ratios (OR) of MD development in representatives of the Yakut population for these three loci were 2.59 (95% CI, p = 0,004), 4.99 (95% CI, p = 0.000), and 3.15 (95% CI, p = 0.01), respectively. Haplotype TTTCTC, which is associated with MD, and haplotype GTCCTT, which was observed only in Yakut MD patients (never in MD patients of non-Yakut origin), were revealed. A low level of variability in the locus of DMRK gene in Yakuts (H(e) = 0.283) compared with other examined populations was noted. An analysis of pairwise genetic relationships between populations revealed their significant differentiation for all the examined loci. In addition, a low level of differentiation in territorial groups of Yakut populations (F(ST) = 0.79%), which was related to the high subdivision of the northern Eurasian population (F(ST) = 11.83%), was observed.

[Replicative study of susceptibility to childhood-onset schizophrenia in Kazakhs].

This paper reports the results of replicative analysis of associations of 15 SNPs in a region of 14 genes previously identified in genome-wide association studies (GWAS) with early-onset schizophrenia in Kazakhs. An association of early-onset schizophrenia with genetic markers in three genes (VRK2, KCNB2, and CPVL) was found. An association of rs2312147 in the VRK2 gene with schizophrenia was also previously reported in the Chinese population, so this marker may be considered as possibly race-specific. Two groups consisting of four and six genes demonstrating intergenic epistatic interactions were revealed by multifactor dimensionality reduction methods. The gene ontologies of 14 studied genes were reduced to variants of one molecular function (peptidase activity) and one biological process (positive regulation of biosynthesis processes). Bioinformatic analysis of the protein-protein interactions of products of the genes under study demonstrates that the products of six out of 14 genes may be involved in a single interrelated network, the major connecting link of which is represented by their ubiquitination by the UBC protein.

[OPTION OF THE TRANSPLANT URINARY TRACT RECONSTRUCTION WITH RECIPIENT'S OWN URETER IN CASE OF THE RENAL TRANSPLANT PATHOLOGY].

This article describes an extremely rare case of reconstruction of allograft ureteropelvic junction by ureter of contralateral kidney. The authors present a concise literature review of this pathology treatment options.

[Replicative association analysis of genetic markers of cognitive traits with Alzheimer's disease in a Russian population].

Replicative association analysis of Alzheimer's disease (AD) with 15 genetic markers associated with cognitive traits in genome-wide association studies was performed. In a Russian populations associations of rs2616984 in CSMD1 gene with AD (OR = 1.50, 95% CI = 1.07-2.09, p-value = 0.018) and putative associations with the disease of rs3131296 in NOTCH4 gene (OR = 1.53, 95% CI = 0.98-2.39, p-value = 0.06) and rs2229741 of NRIP1 gene (OR = 1.35, CI = 0.99-1.85, p-value = 0.061) were revealed. Combinations of epistatic interacting genes (CSMD1 and NRIP1; NOTCH4, CSMD1 and NRIP1; TLR4, CSMD1 and NRIP1) were found, as well as their genotypes combinations significantly associated with AD and characterized by highest predictive values. Probable molecular mechanisms implicated in the relation of genes under study to AD pathogenesis are discussed. Bioinformatic analysis of biological processes, molecular functions and protein-protein interactions of BA genes demonstrated that genes under study may play modulating and modifying role by participation in various regulatory and signal pathways involved in a disease development.

[Association of the genenetic polymorphism of cytokines and their receptors with climate and geographic factors in human populations].

The variability of eight polymorphic variants of the IL4, IL4R, IL10, IL13, IL12A, and IL12RB2 genes encoding key cytokinesand their receptors in 57 world populations has been assessed. A correlation between the allele frequency distribution of the examined genes and climatic and geographic factors was observed.