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Progress in understanding of structural and functional human genome organization and deciphering primary DNA sequence in human cells allowed for hitherto unreachable new capabilities of medical genetics in identifying the causes and mechanisms of inherited and inborn pathology. Implementation of genetics into medicine is progressively advancing along with improvement of molecular analysis of genome. Knowledge of genome and its functions allows to provide more accurate diagnosis, predict, to a considerable extent, the presence of genetic predisposition of a person to pathology, and to assess the chances for developing one or another disease. This approach became the basis for a new area of medical genetics named predictive medicine. The progress of predictive medicine refl ects success in tremendous upgrowth of molecular genetic methods and new capabilities of studying structure and functions of genome. Within less than 15 years after deciphering genome, medical genetics has travelled a long way from a single gene analysis to whole genome studies, from screening of genetic associations to systems genetics of multifactorial diseases, from translational to high-precision genetics, and from genetic passport idea to electronic genetic health records. The development of a genetic passport, prognostic genetic testing, and genomic chart of reproductive health is especially relevant for current practical medicine.

By means of the method of polimerase chain reaction the frequency of blank allele of genes GSTP1, GSTM1, and the frequency of polymorphous alleles of gene GSTP1 in 40 pairs with habitual noncarrying of pregnancy at early stages were studied. The results of the investigation testify to the high rate of gomozygotes GSTT1 0/0 in pairs with habitual noncarrying of pregnancy in comparison with indices in general population of North-West region of Russia (37,7% and 23,3% correspondingly). The genes GSTM1, GSTT1, GSTP1 code enzymes, which take part in the second phase of xenobiotic detoxication system. Taking into consideration that protein products of these genes to significant extent determine the effectiveness of 2nd phase of detoxication (neutralization of xenobiotics by glutathione binding), we are to suppose that unfavorable environmental factorsplay the important role in etiology and genesis of habitual noncarrying of pregnancy at early stages.

Here, we determined basic differentially expressed genes in endometrial lesions compared to eutopic endometrium of the patients with endometriosis (EM). Endometrial biopsy and tissue sampling were performed in ten women with pelvic EM and nine controls at their mid-secretory phase. NGS of mRNA combined with bioinformatic analysis and complemented by RT-qPCR analysis were used for analysis. A list of differentially expressed genes of ectopic compared to eutopic endometrium during mid-secretary phase has been composed. Functional annotation has found profound gene expression perturbations in numerous EM pathways including extracellular matrix, complement and coagulation, as well as cell cycle-related gene family. The genes of alcohol dehydrogenase 1B (ADH1B) and fatty binding protein 4 (FABP4) were determined as the most dramatically up-regulated genes in ectopic endometrial lesions. Both genes were not reported before to be associated with EM. Several other genes including PLA2G2A were also up-regulated in ectopic EM lesions. Notably, FABP4 and ADH1B are key genes in basic metabolism and known for their function in adipose tissue. The results provide intriguing new avenue for endometriosis studies.

Current problems and some future perspectives of molecular genetic study of connective tissue diseases (CTDs) are reviewed. The corresponding clinical manifestation and mode of heritable disorders of connective tissue (HDCT) inheritance are subdivided into inherited, mostly monogenic, forms and mixed or multifactorial dysplasia of connective tissue (MDCTs). Both forms, especially MDCT, pose significant problems for precise clinical diagnostics. Implementation of modern DNA technologies includes new generation sequencing and clinical exome sequencing which provided a substantial impact in understanding the complex molecular genetic background of CTDs. Owing to new technologies, hundreds of new causative genes of CTDs were found and many commercial gene panels were designed for more precise and objective diagnostics of different CTDs. The results of implementation of NGS for analysis of both HDCTs and especially MDCTs complicated with common syntropic diseases as well as some new data on epigenetic causes of CTDs are briefly summarized. Special attention is paid to biological models and cell culture technologies complemented results of NGS in CTDs. Obvious advantages, relevant problems, and definite limitations in clinical implication of NGS for CTDs studies are discussed.

Summary: In this review, we summarize data on 5-hydroxymethylcytosine, 5-formylcytosine and 5-carboxylcytosine – cytosine modifications which are produced by TET-mediated oxidation of 5-methylcytosine in DNA. We show the biochemistry of modified cytosine as well as methods for its global and location analysis. We also highlight the milestones in the evolution of ideas on the biological role of 5-hydroxymethylcytosine, 5-formylcytosine and 5-carboxylcytosine in the mammalian genome since their discovery in 2009 till present.

5-hydroxymethylcytosine (5hmC) is an oxidative derivative of 5-methylcytosine (5mC). Recent studies have revealed a sharp difference in the levels of 5hmC in 2 opposite DNA strands of a given chromosome and a chromosome-wide cell-to-cell variability in mammalian cells. This asymmetric 5hmC distribution was found in cultured cells, which may not fully mimic in vivo epigenetic processes. We have checked whether inter-chromosome and inter-cell variability of 5hmC patterns is typical for noncultured human cells. Using indirect immunofluorescence, we analyzed the localization of 5hmC and its co-distribution with 5mC on direct preparations of mitotically active cells from human embryonic lung and chorionic cytotrophoblast samples. We demonstrated 3 types of chromosomes according to the 5hmC accumulation pattern: hydroxymethylated (5hmC in both sister chromatids), hemihydroxymethylated (5hmC in only 1 sister chromatid), and nonhydroxymethylated ones. Each accumulation type was not specific to any particular chromosome, resulting in different 5hmC patterns between homologous chromosomes, among chromosomes within each metaphase plate, among metaphases in one tissue, and between the tissues. The 5mC distribution was stable: chromosomes were methylated in R-bands and, especially in embryonic lung cells, in the heterochromatic regions 1q12, 9q12, and 16q11.2. Our results provide the first evidence of inter-cell and inter-chromosome variability of 5hmC patterns in human noncultured embryonic and extraembryonic cells.

// Olga A. Efimova 1,2,* , Anna A. Pendina 1,2,3,* , Andrei V. Tikhonov 1,2,3,* , Sergey E. Parfenyev 2 , Irina D. Mekina 1 , Evgeniia M. Komarova 1 , Mariia A. Mazilina 1,2 , Eugene V. Daev 2 , Olga G. Chiryaeva 1,4,5 , Ilona A. Galembo 3 , Mikhail I. Krapivin 2 , Oleg S. Glotov 1,2 , Irina S. Stepanova 6,7 , Svetlana A. Shlykova 8 , Igor Yu. Kogan 1 , Alexander M. Gzgzyan 1,2 , Tatyana V. Kuznetzova 1,2 and Vladislav S. Baranov 1,2 1 D.O. Ott Research Institute of Obstetrics, Gynecology and Reproductology, St. Petersburg, Russia 2 St. Petersburg State University, St. Petersburg, Russia 3 Center for Medical Genetics, St. Petersburg, Russia 4 St. Petersburg State Pediatric Medical University, St. Petersburg, Russia 5 S.M. Kirov Military Medical Academy, St. Petersburg, Russia 6 Institute of Cytology RAS, St. Petersburg, Russia 7 Aimed Clinic, St. Petersburg, Russia 8 AVA-Peter Clinic, St. Petersburg, Russia * These authors have contributed equally to this work Correspondence to: Olga A. Efimova, email: // Keywords : 5-hydroxymethylcytosine, semen quality, sperm DNA fragmentation, human spermatogenesis, testicular spermatogenic cells, Pathology Section Received : August 01, 2016 Accepted : May 21, 2017 Published : June 01, 2017 Abstract We performed immunofluorescent analysis of DNA hydroxymethylation and methylation in human testicular spermatogenic cells from azoospermic patients and ejaculated spermatozoa from sperm donors and patients from infertile couples. In contrast to methylation which was present throughout spermatogenesis, hydroxymethylation was either high or almost undetectable in both spermatogenic cells and ejaculated spermatozoa. On testicular cytogenetic preparations, 5-hydroxymethylcytosine was undetectable in mitotic and meiotic chromosomes, and was present exclusively in interphase spermatogonia Ad and in a minor spermatid population. The proportions of hydroxymethylated and non-hydroxymethylated diploid and haploid nuclei were similar among samples, suggesting that the observed alterations of 5-hydroxymethylcytosine patterns in differentiating spermatogenic cells are programmed. In ejaculates, a few spermatozoa had high 5-hydroxymethylcytosine level, while in the other ones hydroxymethylation was almost undetectable. The percentage of highly hydroxymethylated (5-hydroxymethylcytosine-positive) spermatozoa varied strongly among individuals. In patients from infertile couples, it was higher than in sperm donors ( P

Uterine leiomyoma (UL) is the most common benign tumor in women of reproductive age. Gene therapy using suicidal genes appears to be a promising approach for UL treatment. One of key factors for success of gene therapy is the right choice of genetic construct carrier. A promising group of non-viral carriers for cell delivery of expression vectors is cationic Cys-flanked peptides which form tight complexes with DNA due to electrostatic interactions and the presence of interpeptide disulfide bonds. The paper reports a comparative study of the physico-chemical, toxic, and transfectional properties of the DNA-peptide complexes obtained by matrix polymerization or oxidative polycondensation of Cys-flanked peptides using the chain growth terminator 2-amino ethanethiol. We have demonstrated the therapeutic effect of the delivery of the pPTK-1 plasmid carrying the herpes simplex virus type 1 (HSV-1) thymidine kinase gene into PANC-1, and HEK-293T cell culture as well as into primary UL cells. It has been shown that the carriers obtained by oxidative polycondensation transform primary UL cells more efficiently than those produced by matrix polymerization. Treatment with ganciclovir resulted in the death of up to 40% of UL cells transfected with the pPTK-1 plasmid. The perspectives of use of the polyR6 carrier produced by oxidative polycondensation as a tool for the development of modular peptide carriers for the purposes of UL gene therapy were discussed.

The present study reports on the frequency and the spectrum of genetic variants causative of monogenic diabetes in Russian children with non‑type 1 diabetes mellitus. The present study included 60 unrelated Russian children with non‑type 1 diabetes mellitus diagnosed before the age of 18 years. Genetic variants were screened using whole‑exome sequencing (WES) in a panel of 35 genes causative of maturity onset diabetes of the young (MODY) and transient or permanent neonatal diabetes. Verification of the WES results was performed using PCR‑direct sequencing. A total of 38 genetic variants were identified in 33 out of 60 patients (55%). The majority of patients (27/33, 81.8%) had variants in MODY‑related genes: GCK (n=19), HNF1A (n=2), PAX4 (n=1), ABCC8 (n=1), KCNJ11 (n=1), GCK+HNF1A (n=1), GCK+BLK (n=1) and GCK+BLK+WFS1 (n=1). A total of 6 patients (6/33, 18.2%) had variants in MODY‑unrelated genes: GATA6 (n=1), WFS1 (n=3), EIF2AK3 (n=1) and SLC19A2 (n=1). A total of 15 out of 38 variants were novel, including GCK, HNF1A, BLK, WFS1, EIF2AK3 and SLC19A2. To summarize, the present study demonstrates a high frequency and a wide spectrum of genetic variants causative of monogenic diabetes in Russian children with non‑type 1 diabetes mellitus. The spectrum includes previously known and novel variants in MODY‑related and unrelated genes, with multiple variants in a number of patients. The prevalence of GCK variants indicates that diagnostics of monogenic diabetes in Russian children may begin with testing for MODY2. However, the remaining variants are present at low frequencies in 9 different genes, altogether amounting to ~50% of the cases and highlighting the efficiency of using WES in non‑GCK‑MODY cases.

We studied the effect of peptide AEDG on telomere length and mitotic index of PHA-stimulated blood lymphocytes from young (18-22 years, N=5) and middle-aged (49-54 years, N=6) men. In the younger age group, no significant changes in the mitotic index were detected, while in the middle-aged group, a decrease in this parameter was found in one case. The relative length of telomeric regions of metaphase chromosomes was evaluated by in situ fluorescence hybridization with DNA probes specific to telomeres. After incubation with peptide AEDG, significant changes in the relative telomere length were found in 7 of 11 individuals (3 cases in the younger age group and 4 cases in the middle age group). Significant increase in telomere length after exposure to peptide AEDG was revealed in 5 cases, including two individuals of the younger age group (by 41 and 55%) and three individuals of the middle age group (by 156, 18, and 76%). In one individual of the younger age group and in one of the middle-age group, a significant decrease in telomere length (by 37 and 15%, respectively) was found. A tendency to normalization of telomere lengths was noted: this parameter increased in individuals with initially lower telomere length relative to the group mean value and decreased in individuals with initially longer telomeres compared to the mean length in the group.

Uterine leiomyomas (ULs) are common benign tumors affecting women of different ethnicities. A large proportion of UL has mutations in MED12. Multiple and solitary ULs usually manifest with different severities, suggesting that their origin and growth pattern may be driven by different molecular mechanisms. Here, we compared the frequency and the spectrum of MED12 exon 2 mutations between multiple (n=82) and solitary (n=40) ULs from Russian patients. Overall, we detected MED12 exon 2 mutations in 51.6% (63/122) of ULs. The frequency of MED12 exon 2 mutations was almost two-fold higher in samples from the multiple UL patients than in those from the solitary UL patients - 61% (50/82) versus 32.5% (13/40). The increased MED12 exon 2 mutation frequency in the multiple ULs was not accompanied by significant alterations in the spectrum of mutation categories, which included missense mutations, deletions, splicing defects, and multiple (double/triple) mutations. Each mutation category had a unique mutation set, comprising both frequent and rarely encountered mutations, which did and did not overlap between the studied groups, respectively. We conclude that in contrast to the solitary ULs, the multiple ULs predominantly originate through MED12-associated mechanisms. The nature of these mechanisms seems to be similar in solitary and multiple ULs, as they contain similar mutations. In multiple UL patients, they are likely to be nonsporadic, indicating the existence of specific factors predisposing to multiple UL development. These data suggest that to clearly understand UL pathogenesis, solitary and multiple tumors should probably be analyzed as separate sets.

Aims To study the possible association of catechol-O-methyltransferase ( COMT) Val158Met polymorphism with multiple and solitary uterine leiomyomas (ULs) and to check whether the COMT Val/Val genotype is associated with MED12 exon 2 mutations in fibroids. Methods The COMT Val158Met allele and genotype frequencies were compared between age-matched women with ULs (n=104) and controls (n=59). Patients with UL were subcategorised by diagnosis of solitary (n=59) or multiple (n=45) fibroids and by the presence of somatic MED12 exon 2 mutations in at least one fibroid (n=32) or in neither fibroid (n=26). The association of COMT Val/Val genotype with the presence of any ULs, solitary/multiple ULs and ULs positive/negative for MED12 exon 2 mutations was evaluated by χ 2 tests using a dominant genotype model (G/G vs G/A+A/A) and expressed as ORs and 95% CIs. Results The COMT Val/Val genotype frequency did not differ between the patients with UL and the controls (28.8% vs 18.6%, p=0.149, OR 1.77; CI 0.81 to 3.86). However, it was significantly higher in the patients who had multiple UL compared with the solitary UL (40% vs 20.3%, p=0.028, OR 2.61; CI 1.09 to 6.24) and to the controls (40% vs 18.6%, p=0.016, OR 2.91; CI 1.20 to 7.06). No association of the COMT Val/Val genotype with UL-specific MED12 exon 2 mutations was found (p=0.662, OR 0.77; CI 0.23 to 2.53). Conclusions Women with COMT Val/Val genotype are at high risk of developing multiple uterine fibroids either positive or negative for MED12 exon 2 mutations. These data are important to design new strategies for UL prophylaxis and treatment.

This paper presents an overview of the most significant discoveries and findings in gerontology for 2011–2014. It considers results of the whole genome association studies (GWAS), obtained in a representative cohort of centenarians, protective mutations of Alzheimer’s disease, new data on the role of the hypothalamus as a command center of human aging, results of experimental studies of the geroprotective and anticarcinogenic action of rapamycin and metformin, and meta-data indicating a negative effect of antioxidants in the treatment of lung cancer. Special attention is paid to the problems of aging epigenetics and epigenetic mechanisms of gene activity regulation. Data on the long-term epigenetic effects that affect stable reprogramming of the genome at an early age and clinical observations and experimental data demonstrating the transgenerational transmission of epigenetic changes in the parents to their offspring are examined. There has been a tendency emerging in recent years to the transfer from studies of the dormant genome (anatomy of the genome) to the study of the dynamic genome (its physiology), which opens up new opportunities to clarify the underlying mechanisms of aging and ways of maintaining active longevity.

Background. HLA-G is a non-classical MHC class Ib molecule predominantly expressed in cytotrophoblasts and under pathological conditions also in chronically inflamed and in malignant tissues. Polymorphic sites present in coding and non-coding regions of the HLA-G gene may potentially affect all of these biological features. Materials and methods: Using the PCR method we have studied polymorphism of HLA G gene (alleles G*0101-G0107, -725C/G , 3741del/ins14.) in 118 people aged from 20 till 40 years of the North-West Region of Russia (St. Petersburg). Results: To determine the common alleles of the HLA-G gene the system of allele-specific PCR primers was elaborated. We have determined HLA-G allele distribution in North-west Region of Russian Federation. The minor allele frequencies varied in wide ranges - from 0 % to 39 %. The genetic distances among the Spanish, Chinese, Japanese, Danish, Indian, African Shona, Brazilian, Polish, German and North-west Region of Russian Federation were calculated. Conclusion: The evaluation of alleles rates in North-west of Russian Federation and in populations of other countries revealed the similarity in alleles G*0101-G0107 and polymorphism 3741del/ins14 distribution among the Danish, German and North-west Region populations.

The present review summarizes data on 5-hydroxymethylcytosine - a modification of cytosine with a recently discovered epigenetic effect. The biochemical mechanisms of 5-hydroxymethylcytosine formation and further modification in the mammalian genome are discussed; the role of 5-hydroxymethylcytosine in the epigenetic reprogramming during mammalian gametogenesis and early embryogenesis as well as in the regulation of gene expression is analyzed; data on the diseases and the adverse environmental factors, linked to the DNA hydroxymethyaltion disruptions is shown.

We present a comparative analysis of the allelic polymorphisms of the matrix metalloproteinase (MMP) gene family, including MMP3 (rs3025058), MMP7 (rs11568818), MMP9 (rs17576, rs2250889), MMP12 (rs2276109), and MMP13 (rs2252070), in patients with external genital endometriosis (EGE) and in a control group of healthy women proven to be free of disease by laparoscopic inspection. We found significant differences in the incidence of particular MMP3 and MMP9 alleles, which substantiate the role of matrix metalloproteinases in EGE pathogenesis. We used the Multifactor Dimensionality Reduction (MDR) analysis to show that 14 allelic combinations of the MMP containing MMP3 (rs3025058) × MMP7 (rs11568818) × MMP9 (rs17576) alleles showed a statistically significant association with an increased risk of EGE, while 10 other combinations correlated with a reduced risk of the disease. MDR analysis produced two statistically significant models for MMP allelic combinations involved in EGE progression, both with 100% penetrance and 83 and 78% accuracy.

To compare the frequency and the spectrum of karyotype abnormality in the first trimester miscarriages in women aged under and over 35 years, who conceived naturally (NC) and who conceived through in vitro fertilization (IVF). Comparative analysis of cytogenetic data obtained by karyotyping of miscarriages in patients who conceived naturally, and who conceived through IVF. Patients were subcategorized by their age

Вy PCR-RFLP procedures in groups of children of the Northwest region of Russia we investigated associations of some variants of CYP2J2, CYP4A11 and PTGIS genes participating in a metabolism of arachidonic acid and variants of CDH13, MTHFR, CDKN2BAS genes, and also an extra gene molecular marker rs11191548 T/C revealed on the basis of the GWAS method as candidates of risk of arterial hypertension, with development of a hypertension. The association of these markers with development of a hypertension in children didn't prove to be true.

ACGH is a modern and highly efficient approach for full-genomic scanning of wide spectrum of chromosomal disbalance. Here, we discuss diagnostic capabilities of aCGH and give examples of diagnostic cases.

Using high-density microarrays, we analyzed polymorphism of more than 1500 genetic markers associated with risk of a wide range of multifactorial diseases. Based on functional annotation of genes by bioinformatics resources DAVID and GFINDer we selected a group of 31 genes, whose products are associated with the risk of preeclampsia. Population frequencies of alleles and genotypes for the following genes: ACE, ADIPOQ, ADRB2, ADRB3, AGT, APOE, CRP, CTLA4, CYP1A1, CYP2D6, CYP2E1, EDNRA, ESR1, ESR2, F5, HLA-DQA1, HSPA1A, IL1A, IL1RN, IL6, IL6R, LEP, LEPR, LPL, MTHFR, NOS3, PON1, TAP2, TGFB1, TNFA, VEGFA were established. Comparative analysis between the Russian and Central European population groups revealed statistically significant differences in allele and genotype frequencies for 6 genes: CYP2D6, CTLA4, AGT, NOS3, PON1, ADRB2. The data suggest similar basis of genetic risk of vascular diseases in pregnancy in Russian and European populations and may be used for other genetic and epidemiological studies

Famous Free Radical Theory (FRT) of aging, the 50th year anniversary of which is celebrated in 2015 postulates a crucial role of Reactive Oxygen Species (ROS) in aging. Still it is the most robust theory of aging as mitochondria ROS production (mtROSp) correlates well with four principal ''rules" of aging being universal, endogenous, progressive, and deleterious. Vast number of experiments in different species prove mutagenic effect of ROS and their carcinogenic properties. So far, FRT stimulates the search of new pharmaceuticals with antioxidant activity. However, some recent experimental data and clinical findings render doubt to ROS as a principal senescence drivers and come in conflict with original version of FRT. Growth stimulating effects of ROS and their modest antitumor properties support these objections. One should remember that FRT is only one of the numerous theories of aging. Molecular mechanisms of senescence involve all living systems and numerous metabolic pathways which are also variable owing to the unique properties of individual genome and unique epigenetic modulations operating throughout the lifetime thus making aging a unique private matter. Universal theory of aging that incorporates and explains all known and suggested mechanisms of aging, is illusive. However, knowledge of unique peculiarities of individual genome, its feasible editing and efficient epigenetic regulation of metabolic pathways give a chance to postpone aging and extend period of active longevity.

A genetic certificate is a DNA database reflecting the specific features of a personal genome. The review considers the concept of individual genetic certificates and their significance for prolongation of active longevity and for paving the way to the longest possible lifespan. The involvement of the biological clock and weak link genes in aging processes in human beings is outlined. The significance of genetic tests for allelic polymorphisms and marker genes associated with common multifactorial disorders is stressed. Special attention is paid to the Genome Wide Association Studies technology implemented for analysis of genetic profiles and candidate genes associated with common diseases. Relevant scientific problems are discussed, as well as the public interest in creation of personalized DNA databanks (genetic certificates) for pregnant women, children, athletes, etc. The relationship of the genetic certificate concept to the current international Personal Genome Project is highlighted. The prospects of the introduction of genetic certificates into medical practice are discussed in the context of gerontological problems.

Pre-eclampsia (PE) is a complication of pregnancy that affects 5–8% of women after 20 weeks of gestation. It is usually diagnosed based on the de novo onset of hypertension and proteinuria. Preexisting hypertension in women developing PE, also known as superimposed PE on chronic hypertension (SPE), leads to elevated risk of maternal and fetal mortality. PE is associated with an altered microRNA (miRNA) expression pattern in the placenta, suggesting that miRNA deregulation is involved in the pathogenesis of PE. Whether and how the miRNA expression pattern is changed in the SPE placenta remains unclear. The present study analyzed the placental miRNA expression profile in pregnancies complicated by SPE. miRNA expression profiles in SPE and normal placentas were investigated using an Ion Torrent sequencing system. Sequencing data were processed using a comprehensive analysis pipeline for deep miRNA sequencing (CAP-miRSeq). A total of 22 miRNAs were identified to be deregulated in placentas from patients with SPE. They included 16 miRNAs previously known to be associated with PE and 6 novel miRNAs. Among the 6 novel miRNAs, 4 were upregulated (miR-518a, miR-527, miR-518e and miR-4532) and 2 downregulated (miR-98 and miR-135b) in SPE placentas compared with controls. The present results suggest that SPE is associated with specific alterations in the placental miRNA expression pattern, which differ from alterations detected in PE placentas, and therefore, provide novel targets for further investigation of the molecular mechanisms underlying SPE pathogenesis.

Genotype and allele frequencies of uncoupling proteins 2 and 3 genes (UCP2 and UCP3) and peroxisome proliferator-activated receptors genes (PPARA, PPARD and PPARG) were studied in 206 residents of the siege and in 139 individuals of more than 69 years old (control group). Studied polymorphisms included UCP2 (Ala55Val), UCP3 (C-55T), PPARA (G/C), PPARD (+294T/C), and PPARG (Pro12Ala). The G allele and the G/G genotype (PPARA) were overrepresented in the group of survivors and C/C (UCP3) genotype prevailed in the women of besieged Leningrad compared to relevant control groups of the persons of the same age who did not suffered hungry disaster. Feasible protective effects of PPARA gene allele G and C allele of UCP2 genes are briefly discussed.

We present a comparative analysis of the allelic polymorphism of the matrix metalloproteinase (MMP) gene family, including MMP3 (rs3025058), MMP7 (rs11568818), MMP9 (rs17576, rs2250889), MMP12 (rs2276109), and MMP13 (rs2252070), in patients with external genital endometriosis (EGE) and in a control group of healthy women proven to be free of disease by laparoscopic inspection. We found significant differences in the incidence of particular MMP3 and MMP9 alleles, which substantiate the role of matrix metalloproteinases in EGE pathogenesis. We used the Multifactor Dimensionality Reduction (MDR) analysis to show that 14 allelic combinations of the MMP containing MMP3 (rs3025058) x MMP7 (rs11568818) x MMP9 (rs17576) alleles showed a statistically significant association with an increased risk of EGE, while 10 other combinations correlated with a reduced risk of the disease. MDR analysis produced two statistically significant models for MMP allelic combinations involved in EGE progression, both with 100% penetrance and 83% and 78% accuracy.

We report the sequential changes in 5-hydroxymethylcytosine (5hmC) patterns in the genome of human preimplantation embryos during DNA methylation reprogramming. We have studied chromosome hydroxymethylation and methylation patterns in triploid zygotes and blastomeres of cleavage-stage embryos. Using indirect immunofluorescence, we have analyzed the localization of 5hmC and its co-distribution with 5-methylcytosine (5mC) on the QFH-banded metaphase chromosomes. In zygotes, 5hmC accumulates in both parental chromosome sets, but hydroxymethylation is more intensive in the poorly methylated paternal set. In the maternal set, chromosomes are highly methylated, but contain little 5hmC. Hydroxymethylation is highly region specific in both parental chromosome sets: hydroxymethylated loci correspond to R-bands, but not G-bands, and have well-defined borders, which coincide with the R/G-band boundaries. The centromeric regions and heterochromatin at 1q12, 9q12, 16q11.2, and Yq12 contain little 5mC and no 5hmC. We hypothesize that 5hmC may mark structural/functional genome ‘units’ corresponding to chromosome bands in the newly formed zygotic genome. In addition, we suggest that the hydroxymethylation of R-bands in zygotes can be treated as a new characteristic distinguishing them from G-bands. At cleavages, chromosomes with asymmetrical hydroxymethylation of sister chromatids appear. They decrease in number during cleavages, whereas totally non-hydroxymethylated chromosomes become numerous. Taken together, our findings suggest that, in the zygotic genome, 5hmC is distributed selectively and its pattern is determined by both parental origin of chromosomes and type of chromosome bands – R, G, or C. At cleavages, chromosome hydroxymethylation pattern is dynamically changed due to passive and non-selective overall loss of 5hmC, which coincides with that of 5mC.

Studies on the genetics have focused on the regulation of bone mineral density (BMD) a major predictor of osteoporotic fracture risk. Evidence for a genetic contribution to BMD regulation comes primarily from twin studies thats hawed that between 5070 % of variance in BMD is caused by genetic factors. A great deal of research has been done on gene candidate VDR.3 on 176 postmenopausal women (108 natural menopause and 69 surgical menopause). Polymorphisms of vitamin D receptor have been associated in our group with the speed of bone turnover, registries by DEXA with interval 12 months and DDR test. The every second patient with genotype tt has chance to accompany group with osteoporosis in postmenopausae. In group of women with highs speed of bone turnover frequency t allel in 3 time more than in group with slow bone turnover.

The review of original and literature data dealing with case-reports of women, who used pharmacological reagents during pregnancies. Critical analysis of these data prooves the becessity of strictly individual approach for each woman. Personal recommendation of genetical councellor should be consider the biochemical mechanisms of the drug action,date of pregnancy doses an d duration of consumpyion. The inefficiency and general useless of invasive prenatal karyotyping of the fetus subjected to teratogenic agents are stressed. While repeated (dynamic) ultrasound examinations remains as a single feasible mean to estimate adverse effect of the drug during pregnanacy. Commercially available computerasid programms of teratogenic agents do not exclude routine medical- genetic councelling with subcequent invasive prenatal diagnosis because of the reasons not concerned with drug consumption.

Prenatal diagnosis (PD), as a relatively new area of ​​medical genetics, is experiencing a period of rapid growth. Methodological problems of PD associated with the selection of women at high risk of congenital and hereditary pathology, complex examination of the fetus using non-invasive (ultrasound) and invasive (cytogenetic, molecular, biochemical) research methods have already been resolved. Further increase in the effectiveness of PD, its scientific and practical significance depends entirely on the level of organization of this service in the field and its funding. An important role in the development of modern PD in Russia was played by the PD laboratory of the N.I. BEFORE. Otta RAMS.

The review summarizes the original data concerning involvement of some genes participating in Phase I and Phase 2 of detoxification systems in the origin, progression and treatment efficiency of such common multifactorial diseases as endometriosis, bronchial asthma, alcoholic cirrhosis, habitual miscarriages, gestosis, lung cancer, chronic bronchitis etc. Of special theoretical and practical value of these studies is an identification of highly nonrandom association of particular functionally inferior alleles of the genes belonging to glutathione-S- transferase super-gene family (GSTM1 (0), GSTT1 (0), GSTPi (s) as well as N-acetyl transferase - NAT-2 (s) with these diseases. Thus molecular testing of these alleles might be recommended as feasible predictive test for personal predisposition evaluation, for estimation of remote prognosis and for the treatment strategy in at least some of these diseases. Somewhat about 5-7% of all population in the North-West of Russia possess the most unfavorable combinations of GST genes genotypes (GSTM10/0; GSTTI0/0, GSTPi s/s). Early identification of these individuals by means of genetic screening supplemented with subsequent adequate social and medical care should be considered as feasible social program for efficient prevention of these diseases at the national level. Relying on these data as well as on the polymorphism studies and common mutation identification of other genes the idea of Genetic Form for the pregnant women is suggested and briefly discussed. Implementation of new sophisticated and highly productive molecular techniques amenable for the widespread population gene polymorphism screening becomes highly advisable.

The pregnancy complications and perinatal outcomes in 12 patients with confined placental mosaicism (CPM) were studied. Among them abnormal high level of MShCG (2MoM) was noticed in five patients. Increased resistance in the uterine artery was found at 20 and 36 weeks of pregnancy in women with CPM. Stillbirth was registered in 1/3 of cases; intrauterine growth retardation was detected in j of patients. Morphological signs of the placental insufficiency and villi immaturity were detected in all placentas. Our data reflect a possible negative influence of CPM to the pregnancy outcomes.

The overview represents the recent most conspicuous findings in aging studies. It includes new data on the whole genome association studies (GWAS) in big cohort of centenaries, recently found mutation protecting from Alzheimer disease, discovery of hypothalamus as a command center of human aging, very important data on the negative effect of common antioxidants in the treatment of lung cancer as well as new data concerning antiaging and anticancer effects of common drugs such as rapamycine and metformin. Substantial part of the review is devoted to the epigenetic problems of senescence and feasible impact of basic epigenetic mechanisms (methylation of DNA and histone proteins, DNA heterochromatization) in regulation of gene expression, long-term genome reprogramming during early childhood, and transgeneration transmission of epigenetic traits. The necessity of transition from molecular studies of dormant human genome (anatomy of human genome) to genome in action (dynamic genome) and thus with special emphasis to epigenetic medicine is stressed.

The ratio between the normal (+) and null (0) alleles of the genes encoding glutatione S-transferases M1 (GSTM1) and T1 (GSTT1) were studied in normal individuals from northwestern Russia (control group) and in patients with bronchial asthma (BA). The frequency of the GSTM1 0/0 genotype in the population sample was statistically significantly lower (37.8%) than in the BA patients (82.1%; χ2 = 16.8;P< 0.001; Wχ2 = 15.7; α = 0.01). For the GSTT1 gene, similar data were obtained. The frequency of the GSTT1 0/0 genotype in healthy donors was statistically significantly higher (16.3%) than in the BA patients (73.7%; χ2 = 28.5;P < 0.001; Wχ2 = 23.22; α = 0.01). A significant preponderance of the compound homozygotes for the GSTM1 and GSTT1 null alleles among the BA patients was observed. The frequency of the GSTM1 0/0, GSTT10/0 individuals among the patients was 57.9%, while it was only 4.7% among the controls (χ2 = 27.4; P < 0.001).

Comparative retrospective analysis of serum marker proteins (AFP and hHG), ultrasound examinations and fetal karyotypes was carried out in 306 high risk pregnancies of chromosomal anomalies. Peculiarities of pregnansy histories, birth complications and their outcome were considered respektive of maternal serum hHG values registered at 13-21 weeks of gestation. Statistically significant deviations of maternal serum proteins (MSP) values were registered almost with equal probabilities ih the groups of high risk pregnancies of chromosomal disorders as well as in the women with noncomplicated pregnansy histories (56,7% and 59,3% respectively). Chromosome abnormalities were registered in 29 out of total 306 fetuses subjected to invasive karyotyping. 11 out 20 women with chromosomally abnormal fetuses revealed substantial deviations of MSP values. All cases of chromosomal abnormalities correlated with maternal age (over 35) and ultrasound markers in the foetuses. Elevated values of hHG in maternal blood correlated with unfavourable pregnansy outcome.

Two once-weekly intravenous injections of the polyethylene oxide Polyox WSR-301 (yielding a blood concentration of the order of 5×10−6 g/ml) led to a 38% decrease in the area occupied by sudanophilic lesions in the aortic arch of rats fed an atherogenic diet for two weeks. Perfusion under constant pressure of the formalin-fixed vascular system in the posterior part of the body with physiological saline and then with polyethylene oxide (10−5 g/ml) was without effect in normal rats and in those with mild lipoidosis, but reduced hydrodynamic vascular resistance by 9–14.5% in rats with pronounced lipoidosis. Intravenous injection of polyethylene oxide into anesthetized rats with pronounced lipoidosis in doses that were subthreshold for normal rats (blood concentrations of the polymer were of the order of 10−7 g/ml) caused a 20% decrease in the total peripheral resistance to blood flow, with a 17–20% rise of the blood flow rate in the carotid artery.

A novel phenomenon of unusual selective acridine orange (AO) staining ofpericentromeric heterochromatin regions (HRs) in chromosomal preparations from tissue with known spontaneous mitotic activity (chorionic villi, placenta, embryonic tissues, bone marrow, and testes), as well as embryonic stem cells, is described. Staining with 0.01% AO in a citric-phosphate (pH 5.5) or sodium phosphate (pH 7.0) buffer solution allows the HRs of human chromosomes (1q12, 9q12, 13p11.2, 14p11.2, 15p11.2, 16q11.2, 21p11.2, 22p11.2, and Yq12) and pericentromeric HRs of mouse chromosomes to be reliably detected by the red fluorescence of AO. This method of AO staining does not require any pretreatment. Explanations for metachromatic AO staining of polymorphic pericentromeric HRs in chromosomes of spontaneously dividing cells are suggested. A high reproducibility of the specific AO staining makes it possible to suggest its use as a reliable quick method for detection of polymorphic HRs of human chromosomes in cytogenetic prenatal diagnosis and oncohematology.

The results of detailed phenomenological analysis of self-consciousness disorders as consequences of endogenous psychoses are presented; the authors have examined a large number of patients within the last 15 years. The data have been evaluated in view of their role in rehabilitation. The conclusion has been made that the success of rehabilitation measures through formation of the most adequate self- appraisal in patients with acquired and labile alterations of self-consciousness is more probable in those with manic- depressive psychoses or periodical schizophrenia.

Of 126 families referred for counselling of Duchenne muscular dystrophy (DMD), DNA analysis has been suggested to 119 families with at least one affected child or with an affected close male relative of the woman at risk of being a DMD carrier. A large proportion (about 80 per cent) of the families were represented by sporadic cases (only one affected individual). By means of multiplex polymerase chain reactions with different sets of oligoprimers providing amplification of 10-11 different exons, altogether 49 dystrophin gene deletions were identified (41 per cent). Eighteen deletions clustered in the 5' 'hot spot' region of DMD cDNA and 36 in the distal half of the central rod domain around exons 43-53. An unusually high frequency (18 per cent) of deletions involving exons 17-19 was discovered. Large deletions extending through both 'hot spot' regions and thus occupying over 30-40 exons were recorded in five cases (10 per cent). Seventy-six of 94 families were found to be informative by RFLP analysis for intragenic or extragenetic DNA probes. Carrier status was ascertained in 20 and rejected in 32 female relatives in 40 DMD families. Eight DMD-affected fetuses were diagnosed prenatally by direct deletion testing or by RFLP analysis. Feasible interpopulation variations in the dystrophin gene deletion pattern are discussed. The prospects for more effective prenatal diagnosis and carrier detection in high-risk DMD families in Russia are briefly outlined.

Densitometry of cosmonauts following long-duration missions shows reduction of bone mineral density (BMD). On the average, post-flight BMD remains within the normal range and the broad variability of individual BMD values sometimes is qualified as local osteopenia. Individual reactions are typed by similarity of amount and rate of BMD loss. At present, analysis of functionally significant polymorphism of bone metabolism genes is the most effective instrument for diagnostics of susceptibility to osteopenia and osteoporosis. The investigation was aimed to analyze polymorphism of genes of vitamin-D and (VDR) and calcitonin (CALCR) receptors, and of collagen-1 alpha-1-chain (Col1a-1) in candidate cosmonauts and cosmonauts returned from 5 to 7-mo. missions. According to the results of analysis, in the majority of cosmonauts rapid BMD loss correlated with TT genotype by VDR gene but not with genotypes Tt and tt and associated with carriage of incomplete s-allele in the Col1a1 gene. Yet, in several instances high BMD loss rates were personified with carriers of VDR gene alleles (homo- and heterozygote states--tt and Tt) and heterozygote by Col1a1 gene (Ss).

[The multipurpose probe for real-time assessment of behavior reactions of marine mammals and concurrent temperature and noise pollution measurements was subjected to field testing]

The review was made on gene pass conception as individual DNA data bank reflecting unique genetic peculiarities of each human, its major potential contribution in achievements of active longevity and creation most favorable conditions for maximal duration of individual life-span. Participation of major age-regulated genes such as biological clock genes and the weak chain genes in aging processes of humans is briefly outlined. The significance of genetic testing of allelic polymorphisms and marker-genes implicated in common multifactorial disorders is stressed. The problems evoked by genetic results interpretation are mentioned. Special attention is paid to Genome Wide Association Studies (GWAS) technology implemented for analysis of genetic profiles and candidate genes associated with common diseases. Scientific problems and social interests in creation of individually oriented DNA-data banks (Gene Passes) amenable for the pregnant women, children, sportsmen, etc. are discussed. The relationship of Gene Pass idea to the current international genetic program "Personificated Genome" is highlighted. Feasible perspectives for genetic testing and basic contribution of Gene Pass into gerontology practical medical service are reviewed.