RESUMO
The COMT Val158Met polymorphism has been associated with anxiety and affective disorders, but its effect on anxiety-related personality traits varies between studies. Our purpose was to investigate the effect of COMT Val158Met on personality traits from adolescence to young adulthood in a population representative Caucasian birth cohort. Also its association with educational attainment and anxiety and mood disorders by the age 25 were examined. This analysis is based on the older cohort of the Estonian Children Personality Behavior and Health Study (original number of subjects 593). The personality traits were assessed when the participants were 15, 18 and 25 years old. COMT Val158Met had an effect on Neuroticism in females by age 25 (p=0.001, Bonferroni-corrected for five traits), whereas female Val homozygotes scored the highest. In addition, the Conscientiousness scores of subjects with Val/Val genotype were decreasing in time, being the lowest by the age 25 (p=0.006, Bonferroni-corrected for five traits). By the age 25, males with the Val/Met genotype had mainly secondary or vocational education, whereas female heterozygotes mostly had obtained or were obtaining university education. COMT Val158Met was not associated with anxiety or mood disorders in either gender. These results suggest that genes affecting dopamine system are involved in the development of personality traits and contribute to educational attainment.
Assuntos
Logro , Catecol O-Metiltransferase/genética , Personalidade/genética , Polimorfismo de Nucleotídeo Único , Adolescente , Adulto , Transtornos de Ansiedade/genética , Escolaridade , Feminino , Frequência do Gene , Genótipo , Humanos , Estudos Longitudinais , Masculino , Transtornos do Humor/genética , Fatores SexuaisRESUMO
Molecular genetic analysis of ancient human remains are mostly based on mitochondrial DNA due to its better preservation in human skeletons in comparison with nuclear DNA. We investigated mtDNA extracted from human skeletons found in graves in Yakutia to determine their haplotypes and to compare them with lineages of modern populations. Ancient DNA was extracted from fragments of three skeletons of Yakut graves at At-Dabaan, Ojuluun and Jaraama sites (dating XVIII century) and two skeletons of Neolithic graves at Kerdugen site found in central Yakutia (Churapchinsky, Kangalassky and Megino-Kangalassky districts of Yakutia). Five different haplotypes belonging to specific Asian haplogroups were identified. Lineages of mtDNA of Yakut graves belong to haplo-groups C4a, D5a2 and B5b. Our results indicate the continuity of mitochondrial lineages in the Yakut gene pool during the last 300 years. Haplotypes of two humans from Kerdugen site graves belong to haplogroups A4 and G2a/D. We compared these haplotypes with that of 40,000 Eurasian individuals, 900 of them from Yakutia. No exact matches were found in Paleoasian populations of Chukchi, Eskimos, Koryaks and Itelmen. Phylogenetically close haplotypes (+/- 1 mutation) were found in populations of Yakuts and Evenks, as well as in some populations of China, Southern and Western Siberia.
Assuntos
Povo Asiático/genética , DNA Mitocondrial/genética , Filogenia , Povo Asiático/etnologia , Haplótipos/genética , Humanos , Paleontologia , SibériaRESUMO
Two tribal groups from southern India--the Chenchus and Koyas--were analyzed for variation in mitochondrial DNA (mtDNA), the Y chromosome, and one autosomal locus and were compared with six caste groups from different parts of India, as well as with western and central Asians. In mtDNA phylogenetic analyses, the Chenchus and Koyas coalesce at Indian-specific branches of haplogroups M and N that cover populations of different social rank from all over the subcontinent. Coalescence times suggest early late Pleistocene settlement of southern Asia and suggest that there has not been total replacement of these settlers by later migrations. H, L, and R2 are the major Indian Y-chromosomal haplogroups that occur both in castes and in tribal populations and are rarely found outside the subcontinent. Haplogroup R1a, previously associated with the putative Indo-Aryan invasion, was found at its highest frequency in Punjab but also at a relatively high frequency (26%) in the Chenchu tribe. This finding, together with the higher R1a-associated short tandem repeat diversity in India and Iran compared with Europe and central Asia, suggests that southern and western Asia might be the source of this haplogroup. Haplotype frequencies of the MX1 locus of chromosome 21 distinguish Koyas and Chenchus, along with Indian caste groups, from European and eastern Asian populations. Taken together, these results show that Indian tribal and caste populations derive largely from the same genetic heritage of Pleistocene southern and western Asians and have received limited gene flow from external regions since the Holocene. The phylogeography of the primal mtDNA and Y-chromosome founders suggests that these southern Asian Pleistocene coastal settlers from Africa would have provided the inocula for the subsequent differentiation of the distinctive eastern and western Eurasian gene pools.
Assuntos
Etnicidade/genética , Genética Populacional , Filogenia , Ásia Central/etnologia , Ásia Ocidental/etnologia , Cromossomos Humanos Par 21 , Cromossomos Humanos Y/genética , DNA Mitocondrial/análise , DNA Mitocondrial/genética , Europa (Continente) , Frequência do Gene , Variação Genética , Haplótipos , Humanos , Índia , Masculino , Polimorfismo de Nucleotídeo Único , Classe Social , Sequências de Repetição em TandemRESUMO
Y-chromosome variation was analyzed in a sample of 1127 males from the Western Mediterranean area by surveying 16 biallelic and 4 multiallelic sites. Some populations from Northeastern Europe and the Middle East were also studied for comparison. All Y-chromosome haplotypes were included in a parsimonious genealogic tree consisting of 17 haplogroups, several of which displayed distinct geographic specificities. One of the haplogroups, HG9.2, has some features that are compatible with a spread into Europe from the Near East during the Neolithic period. However, the current distribution of this haplogroup would suggest that the Neolithic gene pool had a major impact in the eastern and central part of the Mediterranean basin, but very limited consequences in Iberia and Northwestern Europe. Two other haplogroups, HG25.2 and HG2.2, were found to have much more restricted geographic distributions. The first most likely originated in the Berbers within the last few thousand years, and allows the detection of gene flow to Iberia and Southern Europe. The latter haplogroup is common only in Sardinia, which confirms the genetic peculiarity and isolation of the Sardinians. Overall, this study demonstrates that the dissection of Y-chromosome variation into haplogroups with a more restricted geographic distribution can reveal important differences even between populations that live at short distances, and provides new clues to their past interactions.
Assuntos
Variação Genética , Polimorfismo Genético , Cromossomo Y/genética , África do Norte , Alelos , Europa (Continente) , Haplótipos/genética , Humanos , Masculino , Região do Mediterrâneo , Repetições de Microssatélites , Oriente Médio , Análise Multivariada , Recombinação GenéticaRESUMO
Mitochondrial HVS-I sequences from 10,365 subjects belonging to 56 populations/geographical regions of western Eurasia and northern Africa were first surveyed for the presence of the T-->C transition at nucleotide position 16298, a mutation which has previously been shown to characterize haplogroup V mtDNAs. All mtDNAs with this mutation were then screened for a number of diagnostic RFLP sites, revealing two major subsets of mtDNAs. One is haplogroup V proper, and the other has been termed "pre*V," since it predates V phylogenetically. The rather uncommon pre*V tends to be scattered throughout Europe (and northwestern Africa), whereas V attains two peaks of frequency: one situated in southwestern Europe and one in the Saami of northern Scandinavia. Geographical distributions and ages support the scenario that pre*V originated in Europe before the Last Glacial Maximum (LGM), whereas the more recently derived haplogroup V arose in a southwestern European refugium soon after the LGM. The arrival of V in eastern/central Europe, however, occurred much later, possibly with (post-)Neolithic contacts. The distribution of haplogroup V mtDNAs in modern European populations would thus, at least in part, reflect the pattern of postglacial human recolonization from that refugium, affecting even the Saami. Overall, the present study shows that the dissection of mtDNA variation into small and well-defined evolutionary units is an essential step in the identification of spatial frequency patterns. Mass screening of a few markers identified using complete mtDNA sequences promises to be an efficient strategy for inferring features of human prehistory.
Assuntos
Clima Frio , DNA Mitocondrial/genética , Emigração e Imigração , Frequência do Gene/genética , Gelo , Filogenia , África do Norte , Ásia Ocidental , Europa (Continente) , Marcadores Genéticos/genética , Testes Genéticos , Haplótipos/genética , Humanos , Mutação/genética , Polimorfismo de Fragmento de Restrição , Tamanho da Amostra , Fatores de TempoRESUMO
Clinal patterns of autosomal genetic diversity within Europe have been interpreted in previous studies in terms of a Neolithic demic diffusion model for the spread of agriculture; in contrast, studies using mtDNA have traced many founding lineages to the Paleolithic and have not shown strongly clinal variation. We have used 11 human Y-chromosomal biallelic polymorphisms, defining 10 haplogroups, to analyze a sample of 3,616 Y chromosomes belonging to 47 European and circum-European populations. Patterns of geographic differentiation are highly nonrandom, and, when they are assessed using spatial autocorrelation analysis, they show significant clines for five of six haplogroups analyzed. Clines for two haplogroups, representing 45% of the chromosomes, are continentwide and consistent with the demic diffusion hypothesis. Clines for three other haplogroups each have different foci and are more regionally restricted and are likely to reflect distinct population movements, including one from north of the Black Sea. Principal-components analysis suggests that populations are related primarily on the basis of geography, rather than on the basis of linguistic affinity. This is confirmed in Mantel tests, which show a strong and highly significant partial correlation between genetics and geography but a low, nonsignificant partial correlation between genetics and language. Genetic-barrier analysis also indicates the primacy of geography in the shaping of patterns of variation. These patterns retain a strong signal of expansion from the Near East but also suggest that the demographic history of Europe has been complex and influenced by other major population movements, as well as by linguistic and geographic heterogeneities and the effects of drift.
Assuntos
Variação Genética/genética , Geografia , Idioma , Cromossomo Y/genética , África do Norte , Alelos , Emigração e Imigração , Europa (Continente) , Frequência do Gene/genética , Marcadores Genéticos/genética , Haplótipos/genética , Humanos , Linguística , Masculino , Modelos Genéticos , Oceanos e Mares , Filogenia , Polimorfismo Genético/genéticaRESUMO
We typed 1801 males from 55 locations for the Y-specific binary markers YAP, DYZ3, SRY10831 and the (CA)n microsatellites YCAII and DYS413. Phylogenetic relationships of chromosomes with the same binary haplotype were condensed in seven large one-step networks, which accounted for 95% of all chromosomes. Their coalescence ages were estimated based on microsatellite diversity. The three largest and oldest networks undergo sharp frequency changes in three areas. The more recent network 3.1A clearly discriminates between Western and Eastern European populations. Pairwise Fst showed an overall increment with increasing geographic distance but with a slope greatly reduced when compared to previous reports. By sectioning the entire data set according to geographic and linguistic criteria, we found higher Fst-on-distance slopes within Europe than in West Asia or across the two continents.
Assuntos
Evolução Molecular , Variação Genética , Modelos Genéticos , Cromossomo Y/genética , África do Norte , Ásia Ocidental , Repetições de Dinucleotídeos , Europa (Continente) , Genética Populacional , Geografia , Haplótipos , Humanos , Masculino , Repetições de Microssatélites , Modelos EstatísticosRESUMO
The number of previous anthropological studies pointed to very complex ethnohistorical processes that shaped the current genetic structure of Croatian island isolates. The scope of this study was limited to the general insight into their founding populations and the overall level of genetic diversity based on the study mtDNA variation. A total of 444 randomly chosen adult individuals from 32 rural communities of the islands of Krk, Brac, Hvar and Korcula were sampled. MtDNA HVS-I region together with RFLP sites diagnostic for main Eurasian and African mtDNA haplogroups were analysed in order to determine the haplogroup structure. The most frequent haplogroups were "H" (27.8-60.2%), "U" (10.2-24.1%), "J" (6.1-9.0%) and "T" (5.1-13.9%), which is similar to the other European and Near Eastern populations. The genetic drift could have been important aspect in history, as there were examples of excess frequencies of certain haplogroups (11.3% of "I" and 7.5% of "W" in Krk, 10.5% of "HV" in Brac, 13.9% of "J" in Hvar and 60.2% of "H" in Korcula). As the settlements on the islands were formed trough several immigratory episodes of genetically distant populations, this analysis (performed at the level of entire islands) showed greater genetic diversity (0.940-0.972) than expected at the level of particular settlements.
Assuntos
DNA Mitocondrial/genética , Frequência do Gene , Adulto , África , Ásia , Croácia , Emigração e Imigração , Feminino , Geografia , Haplótipos , Humanos , Masculino , População RuralRESUMO
About a fifth of the human gene pool belongs largely either to Indo-European or Dravidic speaking people inhabiting the Indian peninsula. The 'Caucasoid share' in their gene pool is thought to be related predominantly to the Indo-European speakers. A commonly held hypothesis, albeit not the only one, suggests a massive Indo-Aryan invasion to India some 4,000 years ago [1]. Recent limited analysis of maternally inherited mitochondrial DNA (mtDNA) of Indian populations has been interpreted as supporting this concept [2] [3]. Here, this interpretation is questioned. We found an extensive deep late Pleistocene genetic link between contemporary Europeans and Indians, provided by the mtDNA haplogroup U, which encompasses roughly a fifth of mtDNA lineages of both populations. Our estimate for this split is close to the suggested time for the peopling of Asia and the first expansion of anatomically modern humans in Eurasia [4] [5] [6] [7] [8] and likely pre-dates their spread to Europe. Only a small fraction of the 'Caucasoid-specific' mtDNA lineages found in Indian populations can be ascribed to a relatively recent admixture.
Assuntos
DNA Mitocondrial/genética , Etnicidade/genética , Evolução Molecular , Haplótipos/genética , Hominidae/genética , Filogenia , Animais , Ásia , Europa (Continente) , Frequência do Gene , Genética Populacional , Humanos , ÍndiaRESUMO
The bovine papillomavirus type 1 (BPV-1) E2 protein is the master regulator of papillomavirus replication and transcription. We have raised a panel of monoclonal antibodies (MAbs) against the BPV-1 E2 protein and used them to probe the structure and function of the protein. Five MAbs reacted with linear epitopes, and four MAbs recognized conformation-dependent epitopes which mapped within the C-terminal DNA-binding and dimerization domain. MAb 1E2 was able to recognize the replication- and transactivation-defective but not the competent conformation of the transactivation domain of the E2 protein. MAb 5H4 prevented efficiently the formation of E2-DNA as well as E2-dependent E1-E2-origin complexes and also dissociated preformed complexes in a concentration-dependent manner. Cotransfection of several MAbs with the BPV-1 minimal origin plasmid pUCAlu into CHO4.15 cells resulted in a dose-dependent inhibition of replication. Inhibition of replication by MAb 5H4 and the Fab' fragment of 5H4 correlated with their ability to dissociate the E2 protein from the DNA. MAb 3F12 and MAbs 1H10 and 1E4, directed against the hinge region, were also capable of inhibiting BPV-1 origin replication in CHO4.15 cells. However, the Fab' fragments of 1H10 and 3F12 had no effect in the transient replication assay. These data suggest that MAbs directed against the hinge region sterically hinder the inter- or intramolecular interactions required for the replication activity of the E2 protein.
Assuntos
Anticorpos Monoclonais/farmacologia , Papillomavirus Bovino 1/fisiologia , Replicação do DNA , Proteínas de Ligação a DNA/fisiologia , Proteínas Virais/fisiologia , Replicação Viral , Sequência de Aminoácidos , Animais , Anticorpos Monoclonais/imunologia , Células CHO , Células COS , Cricetinae , Proteínas de Ligação a DNA/imunologia , Mapeamento de Epitopos , Feminino , Fragmentos Fab das Imunoglobulinas/farmacologia , Camundongos , Camundongos Endogâmicos BALB C , Dados de Sequência Molecular , Proteínas Virais/imunologiaRESUMO
IgG1 class mouse monoclonal antibodies (MAbs) were produced against human glutathione S-transferase Mu1-1 (GSTMu1-1). Eight MAbs of 16 are able to recognize only the native form of the enzyme; 4 MAbs bind to native and denaturated enzyme, and the remaining 4 can bind only to partially denatured antigen in direct ELISA or Western blot. The antibodies recognizing the native form of the enzyme bind to six different epitopes. Three overlapping epitopes are responsible for specific binding of MAbs to different allelic variants of GSTMu1-1. Three allele-specific antibodies, 2E1, 11F12, and 7D11, bind to GSTM1a monomer and the other two, 1H8 and 3H10, recognize GSTM1b monomer.
Assuntos
Alelos , Anticorpos Monoclonais/imunologia , Glutationa Transferase/imunologia , Isoenzimas/imunologia , Adulto , Animais , Afinidade de Anticorpos , Especificidade de Anticorpos , Western Blotting , Ensaio de Imunoadsorção Enzimática , Epitopos/imunologia , Feminino , Glutationa Transferase/genética , Humanos , Hibridomas/imunologia , Isoenzimas/genética , Fígado/enzimologia , Camundongos , Camundongos Endogâmicos BALB C , Desnaturação Proteica , CoelhosRESUMO
Genetic screening for sequence variants associated with disease is assuming increasing importance in clinical medicine as well as in research. We describe an efficient method for such analyses, comprising a combination of practical features: (1) Amplified DNA samples are analyzed for their ability to serve as templates in standardized allele-specific ligation reactions between oligonucleotide probes; (2) Two allele-specific probes, differentially labeled with either of two lanthanide labels, compete for ligation to a third oligonucleotide (the signal from the two labeled probes can thus be directly compared in a sensitive time-resolved fluorescence detection reaction); and (3) Large sets of analyses are processed in parallel using a 96-pin capture manifold, serving to reduce pipetting steps and the risk of contamination. We present here the basis of the technique and its application to the screening for two common mutations causing cystic fibrosis and alpha 1-antiytrypsin deficiency.
Assuntos
Colorimetria/métodos , DNA Ligases/metabolismo , Variação Genética , Proteínas de Membrana/genética , alfa 1-Antitripsina/genética , Sequência de Bases , Fibrose Cística/genética , Regulador de Condutância Transmembrana em Fibrose Cística , DNA , Humanos , Dados de Sequência Molecular , MutaçãoRESUMO
A high-capacity manifold support with immobilized antigen was developed for the analysis of IgE-mediated immune reactivity in allergic subjects. Using this 96-pronged support, specific antibodies were trapped and detected from large sets of serum samples. We describe the binding of large amounts of antigen onto the expanded surface of the manifold support, permitting efficient identification of allergic individuals.
Assuntos
Alérgenos/imunologia , Especificidade de Anticorpos , Hipersensibilidade/diagnóstico , Imunoensaio/instrumentação , Imunoglobulina E/imunologia , Quelantes , Ensaio de Imunoadsorção Enzimática , Európio , Fluorescência , Humanos , Imunoensaio/métodos , Técnicas Imunoenzimáticas , Imunoglobulina E/sangueRESUMO
Large numbers of samples can be efficiently processed through sequential reaction steps using a 96-pronged support that projects into individual microtiter wells. The support was constructed by creating a porous surface on a disposable polystyrene manifold, with avidin coupled to this greatly expanded surface. The shape and high binding capacity of the device allow the parallel transfer of large sets of reaction intermediates between different binding or enzymatic processing steps. We have applied the support to increase the efficiency of preparative and analytical molecular genetic reactions. The support also reduce the risks of sample mix-up and contamination in applications such as PCR and DNA sequencing.
Assuntos
Biologia Molecular/instrumentação , Avidina , Proteínas de Bactérias , Biotina , DNA/isolamento & purificação , Cinética , Reação em Cadeia da Polimerase , Poliestirenos , Sefarose , EstreptavidinaRESUMO
We have devised a procedure, termed capture PCR (CPCR), that permits the rapid isolation of DNA segments situated adjacent to a characterized nucleotide sequence. In this procedure, a DNA sample is restriction-digested and a linker, comprising two base-paired oligonucleotides, is added to the ends by ligation. Multiple extension reactions are performed using a biotinylated primer derived from the known sequence, permitting the subsequent isolation of extension products on a streptavidin-coated support. The enriched fragments are amplified exponentially using another specific oligonucleotide, hybridizing 3' to the biotinylated primer in combination with one of the linker oligonucleotides, now functioning as a PCR primer. The convenience of CPCR is greatly enhanced by using a novel streptavidin-coated manifold, which is constructed so that it projects into each individual well of a microtiter plate. The procedure permits the simultaneous isolation of fragments from large numbers of DNA samples and minimizes the risk of contamination between reactions. We have applied this method to identify DNA sequences located downstream of known sequences in the human genome. The technique has also been used to identify end fragments of sequences cloned in a yeast artificial chromosome (YAC) vector. The reactions can be initiated directly from yeast colonies and provide access to DNA sequence information for these end fragments in a minimal number of steps. With the aid of the present technique, we have isolated over 100 end fragments from YACs derived from the human X chromosome. Isolated end sequences have been used to order YAC clones into a contig.
Assuntos
DNA/genética , Reação em Cadeia da Polimerase/métodos , Sequência de Bases , Cromossomos Fúngicos , Clonagem Molecular , DNA Fúngico/genética , Estudos de Avaliação como Assunto , Vetores Genéticos , Humanos , Dados de Sequência Molecular , Cromossomo XRESUMO
Two fast methods for the purification of mouse monoclonal antibodies from ascites, fluids using Blue-DEAE and 'thiophilic' adsorbent (T gel) in the FPLC system are described. Blue-DEAE chromatography is useful only for IgG1, IgG2a and IgG2b monoclonal antibody purification. T gel is a satisfactory adsorbent for IgG2b purification. Other IgG subclasses and IgM can also be obtained by simple one-step procedures, but the preparations contain small amount of high molecular weight contaminants.
Assuntos
Anticorpos Monoclonais/isolamento & purificação , Líquido Ascítico/imunologia , Imunoglobulina G/isolamento & purificação , Animais , Cromatografia Líquida , Etanolaminas , Géis , CamundongosRESUMO
Serum alpha 1-antitrypsin (A1AT) allo- and phenotypes (including M1, M2 and M3 alleles) were studied in 99 patients with gastric ulcer (GU) and 56 patients with duodenal ulcer (DU) using agarose isoelectric focusing. The results were compared with the A1AT data of a random population sample of similar genetic background (1422 persons). An increase in M2 allotype and M1M2 phenotype as well as a decrease in Z allotype of A1AT was seen in GU in comparison to DU and the random population. There were no particular clinical features which would distinguish patients with M2 allotype from the remainder of the GU group. However, a trend toward elevated serum pepsinogen I and II levels in patients with M2 allotype was seen. When the pepsinogen levels were compared in the GU patient groups with and without M2 allotype, matched between themselves by the state of the gastric mucosa, a statistically significant difference was revealed between pepsinogen II levels in these two groups. No associations were found between DU and any of the A1AT phenotypes.
Assuntos
Alelos , Úlcera Duodenal/genética , Úlcera Gástrica/genética , alfa 1-Antitripsina/genética , Adulto , Idoso , Úlcera Duodenal/sangue , Úlcera Duodenal/patologia , Mucosa Gástrica/patologia , Humanos , Masculino , Pessoa de Meia-Idade , Pepsinogênios/sangue , Fenótipo , Úlcera Gástrica/sangue , Úlcera Gástrica/patologiaRESUMO
Two techniques for phenotyping the human placental alkaline phosphatase system were developed: high-voltage agarose-gel electrophoresis and thin-layer separator isoelectric focusing on agarose. These methods enabled a more rapid and sensitive phenotyping of all common phenotypes than the traditionally employed starch-gel electrophoresis. An extended polymorphism of placental alkaline phosphatase was revealed by isoelectric focusing. The existence of two suballeles of Pl1 allele and two suballeles of Pl2 allele was postulated.