ZNF527 GENE rs386809049 ANALYSIS IN POPULATION OF UKRAINE.

It was shown that some mutations in a number of zinc finger protein (Znf) genes cause intellectual disability (ID). In our study in two affected siblings with ID exome analysis revealed the homozygous coding sequence (cds) indel rs386809049 in the ZNF527 gene. The c.806_808 deletion CAT and insertion TGTGCA (rs386809049) results in substitution of Pro269 and Tyr270 to Leu, Cys and Asn, located in the interdomain region of Zinc finger protein 527. The analyses of site orthologs revealed that Pro269 and Tyr270 amino acid positions are conserved across mammalian species, indicating that there may be an evolutionarily conserved function. To evaluate the ZNF527 gene involvement in intellectual disability pathogenesis analysis of rs386809049 polymorphism in 300 individuals from general population of Ukraine was performed. The following genotypes distribution was detected: CAT/CAT (67.7%), CAT/TGTGCA (31%) and TGTGCA/TGTGCA (1.3%). As far as we know this is the flirt published data on rs386809049 distribution in the populations. The ZNFS27 TGTGCA (polymorphic) allele frequency was 16.8% and CAT(wild type)--83.2% in the general population of Ukraine. Such a high polymorphic allele frequency allows us to suggest that analyzed rs386809049 polymorphism in ZNF527 gene cannot be the major cause of intellectual disability.

[Comparative analysis of associations of polymorphic genes F2, F5, GP1BA and ACE with the risk of stroke development in Russian and Ukrainian populations].

The comparative analysis of the associations between G20210A polymorphism of F2 gene, G1691A polymorphism of F5 gene, -5T/C polymorphism of gene GP1BA, I/D polymorphism of gene ACE and the risk of development of the stroke in two ethnical samplings--Russian and Ukrainian populations--was conducted. It was shown that the patients of the Russian population with genotype DD have a higher level of the risk of ischemic stroke development (OR = 1.4, 95% CI [1.05; 1.78], p = 0.02), whereas genotypes I/I and I/D are associated with the lower level of risk of ischemic stroke (OR = 0.7, 95% CI [0.56; 0.95], p = 0.02). In the Ukrainian ethnical sampling, differences in distribution of genotypes and alleles frequencies between patients with stroke and healthy persons upon given polymorphic locus are not significant, and I/D polymorphism of gene ACE is not associated with the risk of development of the stroke (OR = 0.8, 95% CI [0.48; 1.32], p = 0.45). The G20210A polymorphism of gene F2, G1691A polymorphism of gene F5, -5T/C polymorphism of gene GP1BA are not associated with the risk of stroke in two ethnical samplings.

[ESR1 gene allelic polymorphism analysis in population of Ukraine].

In this study analysis of single nucleotide polymorhisms A351G, C397T and number of TA-repeats in ESR1 gene among 199 healthy volunteers from Ukraine was performed. Data concerning genotypes and particular alleles of polymorphic variants distribution were obtained. Possible mechanisms of these polymorphic variants role in ESR1 gene transcription and estrogen receptor expression changes are being discussed.

[Clinical genealogical and molecular genetic study of patients with mental retardation].

The results of clinical, genealogical, cytogenetic and molecular genetic studies of 113 patients from 96 families with different forms of mental retardation from Ukraine are presented. This study was held as part of the CHERISH project of the 7-th Framework Program. The aim of the project is to improve diagnostics of mental retardation in children in Eastern Europe and Central Asia through detailed analysis of known chromosomal and gene's aberrations and to find the new gene-candidates that cause mental retardation. All patients have normal chromosome number (46XY or 46XX). The cases with fragile-X syndrome were eliminated using molecular genetic methods. Genome rearrangements were found among 28 patients using cytogenetic analysis, multiplex ligation-dependent probe amplification (MLPA analysis) ofsubtelomeric regions and array-based comparative genomic hybridisation (array CGH screening). In 10 cases known pathogenic CNV's were identified, 11 cases are unknown aberrations; their pathogenicity is being determined. The rest cases are known nonpathogenic gene rearrangements. Obtained results show the strong genetic heterogeneity of hereditary forms of mental retardation. The further studies will allow to identificate genes candidates and certain mutations in these genes that may be associated with this pathology.

Aspects of "antigen-antibody" interaction of chicken infectious bronchitis virus determined by surface plasmon resonance.

Authors performed investigation on "antigen-antibody" interaction of chicken infectious bronchitis coronavirus (IBV) by a method based on the surface plasmon resonance (SPR). Presence of space-size effect related to a difference between antigen and antibody particle sizes has been theoretically grounded and experimentally proven. Herewith, the difference between responses of the SPR-sensor to specific and non-specific interactions is considerably less (up to 6.3 times) than the expected one (8 - 11 times). An impact of functionalization of sensor's sensitive element surface, as well as acidity of buffer solution on the activity of antigen-antibody interaction was studied here. The difference between sensor's responses to specific and non-specific interactions increased two-fold from 200 to 432ang sec due to this treatment. When changing the acidity of analyzed solution from pH7.3 to pH6.8, the corresponding difference between sensor's responses increased by 6.3 times from 194 up to 1235ang.sec. Thus, an impact of space-size effect on interaction between IBV antigen and specific antibody can be considerably (almost in 3 times) decreased by reducing the acidity of used buffer solution. The results of our investigation can be successfully applied to develop new methods for detection of pathogens and specific antibodies using SPR.

[Allelic polymorphism of FMR1 gene CGG-repeat region in patients with impairment of natural and stimulated ovulation].

The frequency of heterozygote carriers of (risk zone, alleles of FMR1 gene (40-47 CGG-repeats) was significantly higher in group of patients with ovarian dysfunction than in control group I. The tendency for higher frequency of those alleles was observed in patients with "poor response" to superovulation induction in IVF cycles. The average number of oocytes and follicles, which was obtained after stimulation of superovulation, was significantly decreased in FMR1 gene "risk zone" alleles carriers compared to patients with normal alleles of FMR1 gene. The average general dosage of exogenous gonadotrophin, necessary for superovulation induction was significantly higher in heterozygote carriers of FMR1 gene "risk zone" alleles than in patients with normal genotype. Thereby, the FMR1 gene "risk zone" alleles can be one of the hereditary susceptibility factors of impairment nature and stimulated ovulation.

[Androgen receptor CAG gene polymorphism in men with azoospermia and oligozoospermia in Ukraine].

The number of CAG repeats of exon 1 of AR gene was determined in a group of 228 infertile males with azoospermia (n = 68) and oligozoospermia (n = 160) as well as in control group (124 proven fathers) by fluorescent polymerase chain reaction amplification followed by fragment analysis on automated fluorescent analyzer "A.L.F-express". The frequency of alleles with GAG-repeats < or = 18 was significantly higher (P < 0,01) in the group of patients with azoospermia (17,7%) comparing with the control group (2,4%) as well as in the group of patients with oligozoospermia (12,5%) comparing with the control group (2,4%). The frequency of alleles with CAG-repeats > or = 28 significantly differed (P < 0,01) between the group of patients with oligozoospermia (12,5%) and the control group (2,4%). Our data suggest an association between CAG repeats number and impaired spermatogenesis in azoospermic and oligozoospermic males.

[TGFBI gene mutations in the Ukrainian patients with inherited corneal stromal dystrophies].

Mutations Arg124Cys, Thr538Arg, Arg555Thr, Arg555Gln, Leu558Pro, and His626Arg in TGFBI gene were analyzed by polymerase chain reaction and restriction in 84 patients with various forms of corneal stromal dystrophies from 49 unrelated families and 29 clinically healthy relatives of these patients. A new mutation in TGFBI gene, Leu558Pro, was identified in the patients with atypical lattice dystrophy. The haplotypes of four microsatellite markers surrounding TGFBI gene region were analyzed in 22 families. The data on association of genotype and phenotype suggest that the analysis of TGFBI gene mutations is important for differential diagnostics of corneal dystrophies.

[Molecular-genetic analysis of natural and stimulated ovulation impairment].

The influence of FMR1, INHalpha1, NAT2, GSTT1 and GSTM1 genes on ovarian function, and their association with POF and "poor response" to exogenous GT after ovulation stimulation were investigated. The carriers of Ala257Thr transition predominated in the studied "poor responders" group. This transition combined with intermediate alleles of FMR1 gene was observed in 1.6% POF patients and 2.5% persons from "poor responders" group but in nobody of the control group. The frequency of deletion in GSTM1 gene in "poor responders" group was significantly higher (p = 0,01) than in normal ovulatory control group. The frequency of Ser680Ser-Ala307Ala polymorphic genotype (22.2%) in "poor responders" group was significantly higher (p = 0.028) than in normal-ovulatory control group (7.7%). The daily dosage of GT in intermediate alleles of FMR1 gene carriers as well in patients with "slow acetylation" NAT2 genotype was significantly higher in comparison to patients without intermediate alleles and patients with "quick acetylation" NAT2 genotype. Quantity of oocytes after ovulation stimulation in women with INHa1 gene Ala257Thr transition was significantly decreased in comparison to patients without such mutation. Further investigations of these genes can play a major role in POF studying and modulation of ovarian response to exogenous GT.

Polymorphic short tandem repeats for PCR-based diagnosis of the Charcot-Marie-Tooth 1A duplication in Ukraine.

Charcot-Marie-Tooth neuropathy (CMT) is one of the most common hereditary disorders, affecting 1:2500 individuals. The major mutation--microduplication of 1.4 megabases in 17p11.2 region, which is responsible for 68-90 % of cases of CMT1, results in CMT1A. In the present article we provide the population genetic study in 52 unrelated non-CMT volunteers from population of Ukraine in three STRs (D17S921, D17S1358 and D17S122) from the 17p11.2 chromosomal region to determine their ability for the CMT1A-duplication detection using STR-PCR method in Ukraine. The informativity for the CMT1A detection in current use STR panel is calculated to be 93,6%. It has been shown that current use STR panel analysis is important for CMT1A duplication detection, early differential diagnosis of CMT including prenatal diagnosis and genetic consulting in high risk families.

[The use of DNA analysis for diagnostics of hereditary premature ovarian failure].

Methods of DNA-analysis of 769G --> A mutations in INHalpha1 gene and CGG-repeats polymorphism in FMRI gene have been developed for creating test-systems for genetically caused forms of premature ovarian failure (POF) diagnostics. The frequency of 769G --> A mutation among women population in Ukraine was established and, by preliminary calculations, makes up 2.8%. Results of analysis of CGG-repeats numbers in FMRI gene in the group of 215 women (oocyte donors) revealed five persons with CGG-repeats numbers, that exceeds the normal one (42 copies). Thus the frequency of persons with allels with high risk of premutation in FMRI gene is 2.3%. The results of our research confirm the actuality of genetic tests of mutations in INHalpha1 and FMR1 genes among the women of reproductive age with the purpose of POF prognosis and prevention the birth of children with fragile X syndrome.

Children of chernobyl cleanup workers do not show elevated rates of mutations in minisatellite alleles.

The disaster at the Chernobyl Nuclear Power Plant in April 1986 was accompanied by the release of large amounts of radioisotopes, resulting in the contamination of extensive regions of the Ukraine, Byelorus and the Russian Federation. Cleanup workers (liquidators) and people living on land contaminated with radioactive materials were most exposed. To assess the genetic effects of exposure to ionizing radiation after the Chernobyl accident, we have measured the frequency of inherited mutant alleles at seven hypermutable minisatellite loci in 183 children born to Chernobyl cleanup workers (liquidators) and 163 children born to control families living in nonirradiated areas of the Ukraine. There was no significant difference in the frequency of inherited mutant alleles between the exposed and control groups. The exposed group was then divided into two subgroups according to the time at which the children were conceived with respect to the fathers' work at the power plant. Eighty-eight children were conceived either while their fathers were working at the facility or up to 2 months later (Subgroup 1). The other 95 children were conceived at least 4 months after their fathers had stopped working at the Chernobyl site (Subgroup 2). The frequencies of mutant alleles were higher for the majority of loci (i.e. 1.44 times higher for CEB1) in Subgroup 1 than in Subgroup 2. This result, if confirmed, would reconcile the apparently conflicting results obtained in the chronically exposed Byelorus population and the Hiroshima-Nagasaki A-bomb survivors.

[Polymorphism of the STR-locus of Y chromosomes in Eastern Slavs in three populations from Belorussia, Russia and the Ukraine]. / Polimorfizm STR-lokusov Y-khromosomy u vostochnykh slavian v trekh populiatsiiakh iz Belorussii, Rossii i Ukrainy.

Allelic polymorphism of five microsatellite loci of the human Y chromosome (DYS19, DYS390, DYS391, DYS392, and DYS393) was analyzed in samples of male populations from Ukraine, Russia, and Belarus (152 subjects in total). The allelic diversity indices (Dg) were determined for all loci; they varied from 0.23 to 0.72. The mean values of this parameter in the Ukrainian, Russian, and Belarussian populations were 0.45, 0.47, and 0.52, respectively. A total of 53 different haplotypes were found in 152 subjects from three populations. The most frequent haplotype was found in 14.5% of the subjects, whereas 35 haplotypes (23%) were each found in only one person. The haplotypic diversity index (Dhp) was 0.94. The genetic distances between the populations studied and some populations of Western and Central Europe were estimated. These data were used to construct a phylogram (tree) of genetic similarity between the populations, which demonstrated that the three Eastern Slavic populations are genetically close to one another and remote from Western European populations.

[An analysis of mutations in the 7th, 10th and 11th exons and of the polymorphism of the 4 nucleotide tandem repeats from the 3' end of the 6th intron of the CFTR gene in families from Ukraine with a high risk of mucoviscidosis]. / Analiz mutatsii v 7, 10, 11-m ékzonakh i polimorfizma chetyrekh nukleotidnykh tandemnykh povtorov 3'-kontsa 6-go introna gena TRBM v sem'iakh s vysokim riskom mukovistsidoza iz Ukrainy.

Mutations in the CFTR gene have been screened in 110 families from Ukraine with high risk of cystic fibrosis. Deletion F508 was found in 121 (55%) of 220 CF alleles. Among the rest mutant alleles (with the absence of delta F508) six other mutations occurring in the 7th, 10th, 11th exons of the CFTR gene were screened. In this way, 5 CF alleles may be characterized: 1 allele with R334W (the 7th exon) 1 with 1677 delTa (the 10th exon), 1 allele with G551D and 2 alleles with R553X (the 11th exon). We have screened 865 healthy donors for F508 from different regions of Ukraine. The frequency of heterozygous carriers in different regions ranged from 1:28 to 1:170. Strong linkage imbalance was found between polymorphic markers of 4 bp tandem repeats from the 3'-end of the sixth intron in the CFTR gene and deletion F508.

[Distribution of the 32 base pair deletion of the CCR5 chemokine receptor gene in different regions of Ukraine]. / Rasprostranenie deletsii 32 p.n. v gene retseptora khemokinov CCR5 v razn'ikh regionakh Ukrain'i.

The results of the analysis of 32 b.p. deletion in the CCR5 macrophage chemokine receptor gene in regional populations of Ukraine are presented. The frequency of this mutation ranged from 9.1% to 11% in the Slavs populations but in the population of Crimea Tartars the frequency of mutation was only 5%.

[Possibilities and prospects of the use of DNA analysis in the diagnosis and prevention of inherited disease in the Ukraine]. / Mozhlyvosti ta perspektyvy vykorystannia analizu DNK dlia diagnostyky ta profilaktyky spadkovykh zakhvoriuvan' v Ukraïni.

The results from molecular genetic analysis of some mutations in the 10th and 11th exons of cystic fibrosis transmembrane regulator (CFTR) gene as well as of deletions in the 8, 17, 19, 43, 50, 60th exons of dystrophin gene in 61 CF-families and 21 DMD-families from different Ukraine regions are presented. It was shown that delta F508 frequency of CF-patients was 59.2%, the frequencies of S5491, G551D and K533X were about 1%. The frequency of delta F508-carriers analysed among 365 healthy donors from different regions of Ukraine was 1:40. The analyzed deletions of dystrophin gene were revealed only among 6 DMD-patients. The associations of analyzed mutations of CFTR gene and DMD-gene with RELP's in 4 loci of chromosome 7 and 2 loci of X-chromosome, respectively, were found. The results of prenatal diagnosis of cystic fibrosis and DMD are presented.

[A population genetics study of the allelic polymorphism in the hypervariable region of the apolipoprotein B gene in the population of different regions of Ukraine]. / Populiatsionno-geneticheskoe issledovanie allel'nogo polimorfizma gipervariabel'noi oblasti gena apolipoproteina B u naseleniia raznykh regionov Ukrainy.

The allelic variation of the hypervariable locus from 3'-untranslated region of apolipoprotein B gene in healthy volunteers from different regions of Ukraine was analysed. Among 396 DNA samples studied, 13 allelic variants were identified with the number of repeats ranging from 29 to 53. The frequency of alleles varied from 0.0013 to 0.3575 with the mean heterozygosity index 77.9%. The investigation of the genetic differences between 4 populations of different regions of Ukraine has been performed. A high level of the polymorphism and a heterozygosity index of APOB locus allow to recommend this polymorphic system as an informative marker for study of genetic structure population and to forensic medical analysis.

[An analysis of the mutations in the mucoviscidosis gene: the origin and distribution of the delF508 major mutation in Ukraine]. / Analiz mutatsii v gene mukovistsidoza: proiskhozhdenie i rasprostranenie mazhornoi mutatsiii delF508 v Ukraine.

Seven mutations in CFTR gene have been screened in patients with cystic fibrosis from Ukraine. The frequency of major mutation delF508 among patients and healthy donors from different regions was studied. Data of age, origin and possible ways of spreading of this mutation in Ukraine have been discussing.

[The polymorphism of the tandem repeats in the dystrophin gene in the Ukrainian population]. / Polimorfizm tandemnykh povtorov gena distrofina v populiatsii naseleniia Ukrainy.

The genetic heterogeneity of populations of different regions of Ukraine was studied using four polymorphic LNA sequences (GTAA-tandem repeats from the 3'-untranslated region of the dystrophin gene, the polymorphic systems pERT87-15--BamHI, pERT8718--TaqI, and pERT87-15--XmnI of the pERT locus of human chromosome X). We analyzed 500 DNA samples of healthy donors from the Kiev, Lvov, Poltava, and Lugansk regions. The above-mentioned polymorphic DNA sequences were used as markers to determine genetic distances between the regional populations of Ukraine. The data obtained indicate the existence of significant genetic differences among the regional populations of Ukraine. DNA polymorphism in the pERT87-8--TaqI system contributes the most significantly to these differences.

[Allele polymorphism of micro- and minisatellite loci in populations of different regions of Ukraine]. / Allel'nyi polimorfizm mikro- i minisatellitnykh lokusov sredi naseleniia regional'nykh populiatsii Ukrainy.

The results of molecular-genetic analysis of 12 mini- and microsatellite loci in populations of different regions of Ukraine (Kiev, Kremenchug, L'vov, Lugansk, and in Crimea tatars) were presented. Allele frequencies for each locus were determined and genetic distances between analyzed populations were calculated. The results of the analysis were applied for investigation of genetic heterogeneity and biological history of populations from different regions of Ukraine.