DISTRIBUTION OF RNA-CONTAINING BEE VIRUSES IN HONEY BEE (APIS MELLIFERA) IN SEVERAL REGIONS OF RUSSIA.

In several regions of Russia, broad distribution of RNA-containing bee viruses was found at apiaries of honey bee Apis mellifera using RT-PCR. Detected RNA-containing bee viruses are transferred simultaneously with invasion of mite Varroa destructor and lead to mass bee mortality that results in economic losses in bee breeding. In samples of Varroa destructor, bee viruses DWV and ABPV were found. High degree of RNA-containing virus (BQCV, DWV SBV, ABPV, CBPV and KBV) infection was revealed: in the average, at least 50% for bee families with mite infection. In the bee families studied in this work, mixed infection with 2-6 viruses simultaneously was detected. Amplified fragments of viruses BQCV, DWV and SBV obtained using RT-PCR were sequenced and registered in Genbank.

[Developing forensic reference database by 18 autosomal STR for DNA identification in Republic of Belarus].

For the Republic of Belarus, development of a forensic reference database on the basis of 18 autosomal microsatellites (STR) using a population dataset (N = 1040), "familial" genotypic dataset (N = 2550) obtained from expertise performance of paternity testing, and a dataset of genotypes from a criminal registration database (N = 8756) is described. Population samples studied consist of 80% ethnic Belarusians and 20% individuals of other nationality or of mixed origin (by questionnaire data). Genotypes of 12346 inhabitants of the Republic of Belarus from 118 regional samples studied by 18 autosomal microsatellites are included in the sample: 16 tetranucleotide STR (D2S1338, TPOX, D3S1358, CSF1PO, D5S818, D8S1179, D7S820, THO1, vWA, D13S317, D16S539, D18S51, D19S433, D21S11, F13B, and FGA) and two pentanucleotide STR (Penta D and Penta E). The samples studied are in Hardy­Weinberg equilibrium according to distribution of genotypes by 18 STR. Significant differences were not detected between discrete populations or between samples from various historical ethnographic regions of the Republic of Belarus (Western and Eastern Polesie, Podneprovye, Ponemanye, Poozerye, and Center), which indicates the absence of prominent genetic differentiation. Statistically significant differences between the studied genotypic datasets also were not detected, which made it possible to combine the datasets and consider the total sample as a unified forensic reference database for 18 "criminalistic" STR loci. Differences between reference database of the Republic of Belarus and Russians and Ukrainians by the distribution of the range of autosomal STR also were not detected, corresponding to a close genetic relationship of the three Eastern Slavic nations mediated by common origin and intense mutual migrations. Significant differences by separate STR loci between the reference database of Republic of Belarus and populations of Southern and Western Slavs were observed. The necessity of using original reference database for support of forensic expertise practice in the Republic of Belarus was demonstrated.

[ASSOCIATION BETWEEN FOUR SEROTONIC GENES POLYMORPHISM (5HTTL, 5HT1A, 5HT2A, AND MAOA) AND PERSONALITY TRAITS IN WRESTLERS AND CONTROL GROUP].

This study presents the data on the polymorphisms of the serotonin system genes (5-HTTL, 5-HT1A, 5-HT2A, and MAOA) in male and female wrestlers and in the control group. The population genetics analysis of the 5HTTL gene showed the highest frequency of the SS genotype 5-HTTLPR in sportsmen (p = 0.04), as well as the trend toward higher frequency of united genotypes of the locus of 5-HTTLPR VNTR and SNP rs25531--SASA (p = 0.06) in comparison with the control group. As for the polymorphisms for other genes 5-HT1A (rs6295), 5-HT2A (rs6311), and MAOA (VNTR), we found no significant differences between the groups tested. Using the NEO PI-R questionnaire we analyzed the possible correlations between the genotypes and the psychological traits in our samples. It was demonstrated that the athletic success in elite sportsmen was associated with lower openness to experience and higher conscientiousness. The interaction effect of the gender and 5-HT2A on the self-rating for openness to experience, interaction effect of the level of the sport success and 5-HT2A, and the interaction effect of the gender and 5-HT1A genotype on self-reported conscientiousness were observed as a trend.

[Genetic demography of populations of three megalopolises in relation to the problem of creating genetic databases].

Based on data collected from urban residents by questionnaire, the basic parameters of the genetic-demographic structure of populations of the three megalopolises, i.e., Moscow, Kharkov, and Minsk, have been calculated, including the migration coefficients and their dynamics in generations, the radius of the cities migration attraction, the parameters of marriage structure (the proportion ofinterethnic marriages, the level of intraethnic assortative mating, the marital distances), and the gene flow between the ethnic groups. It is shown that the representatives of the most numerous ethnic groups in each megalopolis have considerable amount of admixture. For Russians of Moscow, Ukrainians of Kharkov, and Belarusians of Minsk, the proportion of individuals whose ancestors were all born in the given city for at least three generations and belonged to the same nationality turned out to be very low (4.75% in Moscow, 1.83% in Kharkov, and 3.13% in Minsk). This finding questions the formation of a reference population in the megalopolis as a sampling of aboriginals of certain ethnic origins. In the paper, we justify principles of creating genetic databases for the population of the megalopolis taking into account the complexity and dynamism of its population structure.

[Variation of morphophysiological and genetic demographic traits in children with congenital cleft lip and palate].

Medical records and questionnaire data have been used to analyze morphophysiological (the birth weight and length) and genetic demographic (maternal age and marriage structure) traits in a sample of children with orofacial malformations (OMs, cleft palate and/or cleft lip) living in Krasnodar krai, Russia. The sample of children with malformations (including premature infants) differs from the control group in lower birth weight and length and a lower proportion of children with morphophysiological values close to the population average values, as well as a higher family exogamy level estimated on the basis of marriage structure in the parental and preceding generations. The risk of congenital cleft palate and/or cleft lip is considerably increased if the material age is over 35 years or, to a lower degree, if it is under 20 years.

[RT-PCR detection of deformed wing virus in the honey bee Apis mellifera L. in the Moscow Region].

Deformed wing virus (DWV) was first detected in the honey bee Apis mellifera by reverse transcriptase-polymerase chain reaction (RT-PCT) in the Moscow Region. Molecular phylogenetic analysis of the detected nucleotide sequence of the virus fragment VP2-VP1 of DWV demonstrated that the Russian virus sequence is united in the common cluster with all earlier revealed nucleotide sequences of DWV in the Genbank worldwide, which confirms the previous conclusions that this virus has recently distributed in the honey bee by Varroa destructor mite. It has been shown that the level of homology for all DWV nucleotide sequences is 98%, except for nucleoside sequence of 7D isolate from Turkey (96% homology), 96% homology with Kakugo virus and 84-86% homology with Varroa destructor virus 1; there is a preponderance of insignificant nucleotide substitutions, mainly transitions, which supports the evolutionary propinquity of 3 viruses.

[Genetic HLA markers associated with pulmonary tuberculosis in the Barum-Khemchiksky District, Republic of Tyva].

Detection of the genetic markers determining a predisposition to pulmonary tuberculosis is a necessary condition for the warranted formation of risk groups in the populations. On this basis, the authors used immunogenetic studies to examine 60 patients with pulmonary tuberculosis and 96 healthy individuals of Tuvinian nationality, who lived in the Barum-Khemchiksky District, Republic of Tyva. The microlymphocytotoxic test was used to determine class I HLA antigens and polymerase chain reaction was employed to reveal the specificity of class II HLA-DRB1 gene. The study revealed a positive association of HLA-B27 antigen and the specificities of HLA-DRB1 13(6) HLA-DRB1 14(6) with tuberculosis, which permits tuberculosis risk groups to be formed, by taking into account the immunogenetic data obtained in this district of the Republic of Tyva.

[Study of the polymorphic markers--the PARK2 and PACRG genes due to the incidence of pulmonary tuberculosis in two districts of the Republic of Tyva].

Polymerase chain reaction-RFLP was used to distribute two polymorphic markers (SNP) PARK2-e01 (-697) and rs1333955 located at the common promoter site of the PARK2 and PACRG genes in order to reveal assumed associations with the incidence of pulmonary tuberculosis in the Barum-Khemchiksky and Ovyursky Districts of the Republic of Tyva. No significant differences were found in the frequencies of these two polymorphic markers between the groups of patients with tuberculosis and healthy individuals, residing in the above districts, and between the total control samples from both districts. The total group of patients with tuberculosis from the two districts from the Republic of Tyva showed a significant surplus of heterozygotes in both study markers, as compared with the group of healthy individuals, which was also observed for the marker rs1333955 in the Barum-Khemchksky District alone. The observed features of genotypic distribution by the two study markers point to the influence of the considered markers on the incidence of tuberculosis.

[NotI-STS markers for human chromosome 3 are gene markers].

Ninety four NotI-STS markers to seventy two individual NotI clones were developed basing on DNA nucleotide sequences from NotI-"jumping" and "linking" NotI-libraries of human chromosome 3. The localization of NotI-STS markers and their ordering on chromosome was established by combined data of RH-mapping (our data), contig-mapping, cytogenetic mapping and in silico mapping. Performed comparison of NotI-STS DNAs with human genome sequences revealed two gaps in the regions, 3p21.33 (marker NLI-256) and 3p21.31 (NL3-005), and segmental duplication. Identical DNA fragments are localized in the regions 12q and 3p22-21.33 (marker NL3-007). In the region 3q28-q29 (marker NLM-084) a fragment was detected with its identical copies present also on chromosomes 1, 2, 15 and 19. For 69 NotI-STSs, significant homologies with nucleotide sequences of 70 genes and two cDNAs were detected taking in consideration homologies to NotI-STS 5'- and 3'-terminal sequences. Association of NotI-STSs with genes is confirmed by high correlation of gene density distribution with the density of NotI-STS markers on the map of human chromosome 3. Obtained data evidence possibility of NotI-STS marker application as gene markers and allow considering constructed NotI-map as gene map of human chromosome 3.

[Characteristics of the RFLP marker of the D13S25 locus on human chromosome 13: mapping of SspI sites in the pH2-42 probe]. / K voprosu o kharakteristike PDRF-markera lokusa D13S25 13-i khromosomy cheloveka: lokalizatsiia SspI-saitov v probe pH2-42.

Analysis of probe pH2-42 of locus D13S25 in genomic DNA in Moscow population has been made by PCR-RFLP using restriction endonuclease SspI. Localization of additional SspI-site at position 1240 (which has been revealed earlier by sequencing of pH2-42) was confirmed. This SspI restriction site was present in all representatives of the population, thus being not polymorphic. Consequently establishment of identity of additional site at position 1240 with polymorphic SspI site in D13S25 studied in previous investigations, requires additional data from another populations. Possible localization of polymorphic SspI-site in locus D13S25 is discussed.

[Brain-specific sequence Hfb1 is part of the 3'-untranslated region of the human complexin 2 gene: new data from in vitro and in silico experiments]. / Mozgospetsificheskaia posledovatel'nost' Hfb1 vkhodit v sostav protiazhennoi 3'-netransliruemoi oblasti gena komplksina 2 cheloveka: novye dannye in vitro i in silico éksperimentov.

A genomic clone hybridizing with brain-specific sequence Hfb1 was isolated from a chromosome 5 consmid library. Hfb1 proved to correspond to a new gene exon which codes for a large 3'-untranslated region of the mRNA for synaptic protein complexin 2. Together with the 985-nt Hfb1 cDNA (EMBL Y15167) isolated previously from a cDNA library of the frontal cerebral cortex, the primary structure was established for genomic clone Ghfb sized more than 4 kb. A GenBank search revealed complete identity of the 5' end of Ghfb and the 3'-untranslated region (878-933) of the human complexin 2 mRNA. Large transcripts with the 5' end corresponding to the complexin 2 mRNA and the 3' end to Ghfb were detected in total mRNA of the human brain by means of RT-PCR. The size of the 3'-untranslated region of the human complexin 2 mRNA was estimated at 4 kb.

[Characteristics of the pH2-42 probe for the D13S25 locus of human chromosome 13: nucleotide sequence, localization, and PCR markers]. / Kharakteristika proby pH2-42 lokusa D13S25 13-i khromosomy cheloveka: nukleotidnaia posledovatel'nost', lokalizatsiia, PTsR-markery.

The sequence of the HindIII-HindIII fragment of probe pH2-42 of locus D13S25 of human genome is given. Localization of the probe in q14-q21 of human chromosome 13 is confirmed by hybridization in situ. Seven oligonucleotide primers for the polymerase chain reaction are chosen so that amplified products almost completely cover the analyzed sequence. Reconstruction of localization of polymorphic SspI-sites in D13S25 was based on the data of Bowcock and Hebert [3] and this study. The results obtained make it possible to use the primer sets to screen cosmid libraries and to mark the D13S25 locus of human chromosome 13.

[Radiation mapping as one of main methods of creating high resolution maps of human and animal genomes]. / Radiatsionnoe kartirovanie kak odin iz osnovnykh metodov postroeniia fizicheskikh kart genomov cheloveka i zhivotnykh s vysokim urovnem razresheniia.

Radiation hybrid mapping (RH mapping) is considered as one of the main methods of constructing physical maps of mammalian genomes. In introduction, theoretical prerequisites of developing of the RH mapping and statistical methods of data analysis are discussed. Comparative characteristics of universal commercial panels of the radiation hybrid somatic cells (RH panels) are shown. In experimental part of the work, RH mapping is used to localise nucleotide sequences adjacent to NotI sites of human chromosome 3 with the aim to integrate contig map of NotI clones to comprehensive maps of human genome. Five nucleotide sequences adjacent to the sites of integration of papilloma virus in human genome and expressed in the cells of cervical cancer were localised. It was demonstrated that the region 13q14.3-q21.1 was enriched with nucleotide sequences involved in the processes of oncogenesis. RH mapping can be considered as one of the most perspective applications of the modern radiation biology in the field of molecular genetics, that is, in constructing physical maps of mammalian genomes with high resolution level.

[Population genetics of taiga hunters-deer breeders. Characteristics of the prevalence of HLA system markers among the native population of Central Siberia]. / Genetika populiatsii taezhnykh okhotnikov-olenevodov. Osobennosti rasprostraneniia markerov sistemy HLA v korennom naselenii Srednei Sibiri.

The postulated peculiarities of distribution of the HLA antigens in the Evenk population of the Central Siberia indicate this population to be, on the whole, Mongoloid. Prolonged isolated existence of small inbred populations of the Evenks resulted in the number of specific HLA-complex features, such as poor antigen diversity, low level of heterozygosity. Non-homogeneity of gene frequencies was noted, the prevailing genes being A9, B40 with frequencies reaching 50%. Of the remaining alleles found in the Evenks A2, Aw19 in A locus, B5, B15, B17, B27, Bw35 in B locus and Cw2, Cw3, Cw4-in C locus are only widely spread. Similar heterozygosis levels and the HLA-system profiles draw the Evenks and the populations of American Indians and Eskimos closer to one another. On the basis of the fact that significant positive correlation exists between genetic and geographic distances determined for the Evenks and the native populations of America, genetic resemblance of these populations may be considered as evidence for their original relationship. Postulated closeness of native Central-Siberian populations to Americanoids, as regards HLA-system, makes it possible to consider formation of specific complex of HLA-system peculiarities in native American population in the light of their Siberian origin, for the first time.

[Genetic polymorphism of the HLA system in Komi and Komi-Permyaks]. / Geneticheskii polimorfizm sistemy HLA u komi i komi-permiakov.

High frequencies of HLA-A2, -A3, -A9, -B7, -B13, -Bw35, and -Dw3 antigens were found in Sysolian and Udorian Komi (from Vizinga and Koslan, respectively), as well as in Zyuzdian Komi-Permyaks. In some populations, high frequencies of HLA-A10, -A28, -B21, -B27, and -B40 antigens were also found. Similarity in both gene frequency patterns and heterozygosity level was noted between these populations and the population of more western European regions, as well as Finno-Ugric nations. Frequencies of some alleles, e.g., HLA-B8 -B12, -B16, and -B18, were found to be intermediate between the corresponding estimates for Mongoloids and Caucasoids. The Mongoloid influence on the gene pool is expressed in increased HLA-B27, -B40, -Cw2, -Cw3, and -Cw4 frequencies. The frequencies of HLA-A3, -A28, -B7, -B13, and -B35 antigens are higher, and that of HLA-B5 is lower, than the respective frequencies in both Caucasoids and Mongoloids. This reflects the specificity of the ancestral group, whose peculiarities cannot be attributed exclusively to the results of Caucasoid-Mongoloid intermixing. The high level of genetic differentiation of Komi ethnographic groups (F st = 0.0188 +/- 0.0045) is emphasized. The characteristics of distribution of HLA antigens and haplotypes are explained in terms of the ethnogenesis of the Komi; markers for their constituent ethnic components were found.

[Polymorphism of the kappa-casein gene in populations of the subfamily Bovinae]. / Polimorfizm gena kappa-kazeina v populiatsiiakh podsemeistva Bovinae.

Polymorphism of the 5'-untranslated region and exon 4 of kappa-casein (kappa-casein) gene was studied in Yakutian and Black Pied cattle, yak, European bison, and buffalo by means of a polymerase chain reaction and subsequent restriction fragment length polymorphism analysis (PCR-RFLP). In the species studied, restriction polymorphism by the endonucleases AluI and Bg/II in the 5'-untranslated region of the gene is absent. Four restriction endonucleases testing nucleotide substitutions in 136 codon (TaqI), 148 codon (HinfI and HindIII), and 167 and 168 codons (PstI) were used to study polymorphism of exon 4. The use of several restriction endonucleases allowed three alleles of kappa-casein (kappa-CnA, kappa-CnB, kappa-CnF) to be typed and new allele variants in yak, European bison, and buffalo to be revealed. Nucleotide sequences of the fragments of exon 4 studied were determined for two new alleles of the gene: kappa-CnG in yak and European bison and kappa-CnH in buffalo. Nucleotide substitutions determining new alleles were localized. In kappa-CnG, 148 and 168 codons coincide with the corresponding codons of kappa-CnB, and 136 and 167 codons correspond to kappa-CnA. Stop codons of kappa-CnG in yak are different from stop codons of other alleles of the gene: TGA, instead of TAA. The nucleotide sequence of exon 4 of kappa-CnH differs from bovine kappa-CnA by 15 nucleotide substitutions, causing 10 amino acid changes in the protein sequence, which coincide with the corresponding known amino acid sequence of kappa-casein in buffalo. Interbreed and interspecies differences in the profile of allele frequencies of the species studied were revealed. Aspects connected with evolution of the alleles of kappa-casein are discussed.

[Genetic diversity of moose (Alces alces L.) in Eurasia]. / Geneticheskoe raznoobrazie losia (Alces alces L.) v Evrazii.

Polymorphism of nucleotide sequence of D-loop fragment of the mitochondrial DNA was studied in 20 moose from several local populations on the territory of Eurasia. Three main haplotype variants of D-loop were detected by molecular phylogenetic method, which formed three clusters named European, Asian and American. Intraspecies variation in the length of HVSI of D-loop of the mitochondrial DNA of moose was revealed. In the Far Eastern and Yakutian moose, haplotypes with a 75-bp deletion were found, which were most similar with haplotypes (also with the deletion), earlier observed in North American moose [1]. The highest diversity of the haplotypes of mitochondrial DNA is characteristic of Yakutia and the Far East (where three haplotype variants were found), which demonstrates the probable role of the region as the center of the species or as the region of ancient population mixture. The geographic region might be considered as a probable source of ancient moose migrations from Asia to America, basing on the data of distribution of mitochondrial haplotypes of D-loop and alleles of MhcAlal-DRB1. Divergence of nucleotide sequences of haplotypes with the 75-bp deletion (forming the American cluster on the phylogenetic tree) was the lowest (0.4%), which evidences respectively recent origin of the group of haplotypes. In Europe, only haplotypes of mitochondrial DNA referred to European variant were observed. Basing on analysis of variation of nucleotide sequences of D-loop, exon 4 of kappa-Casein and exon 2 of MhcALal-DRB1, we demonstrated that Eurasian moose studied belong to the unique species, which has probably passed through a bottle neck. The time of the origin of modern diversity of D-loop haplotypes of the species was estimated as 0.075-0.15 Myr ago.

[The HLA system of the Uzbek population in the Ferghana Valley. HLA antigens, genes and haplotypes of the Uzbek population in relation to its ethnogenesis]. / Issledovanie sistemy HLA uzbekskogo naseleniia Ferganskoi doliny. HLA-antigeny, geny i gaplotipy uzbekskoi populiatsii v sviazi s ee étnogenezom.

Pecularities of distribution of 40 HLA antigens within the Uzbek population of the Ferghana Valley have been studied. The frequencies of these antigens are subdivided into three main groups having frequencies characteristic of the following populations: Caucasoid population (A9, B40, Bw22 etc.); Mongoloid population (B8, B7, B12); Middle Asian population (Aw31, B13, B16, Bw35), probably. The Uzbek population contains haplotypes both of Europeoid (Aw30, B13; A3, Bw35) and Mongoloid origin (A9, B40) and, probably, of the local origin (A1, B14; A1, Bw53; Aw32, B40). The data on the HLA genetics of the Uzbeks confirm the historical, linguistic and anthropological information concerning the role of inhabitants of the Central Asia in ethnogenesis and formation of the modern Uzbek population.

[Blood group polymorphism in bison (Bison bonasus)]. / Izuchenie polimorfizma grupp krovi u zubrov (Bison bonasus).

Polymorphism of nine blood group systems was studied in three reproductive lines of auroch (European bison Bison bonasus), Belovezhskaya, Caucasian-Belovezhskaya, and Gornaya. All blood group systems analyzed were polymorphic; 50 out of the 57 antigens tested were detected. The number of detected blood group antigens in aurochs is comparable to that in breeds of domestic cattle. However, intrapopulation diversity, estimated with the mu test, is significantly higher in cattle than in aurochs. The lowest mu values were observed in line Belovezhskaya, which was founded by a small number of animals and has a high level of inbreeding. For the FV system, the amount by which heterozygotes exceeded the Hardy-Weinberg proportions was shown; this deviation was significant in all lines except Caucasian-Belovezhskaya. The excess of heterozygotes may be due to population processes that prevent the loss of genetic diversity in populations. The level of differentiation in the auroch lines was low. Similarity coefficients between the lines were higher than those between different stocks within a cattle breed.

[Distribution of the kappa-casein gene alleles in the bison (Bison bonasus) population]. / Raspredelenie allelei gena kappa-kazeina v populiatsii zubra (Bison bonasus).

The polymerase chain reaction method, followed by restriction fragment length polymorphism analysis (PCR-RFLP), was used to study the alleles of the kappa-casein gene in a sample of European bison (n = 38). On the basis of the distribution of the restriction sites, a new allele kappa-CnG was revealed in the European bison, in addition to the kappa-CnA allele represented in cattle. The nucleotide sequence of an exon 4 fragment of the kappa-CnG allele was obtained. In the population of European bison alleles kappa-CnA and kappa-CnG occurred at frequencies 0.224 and 0.776, respectively.