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1.
Tissue Antigens ; 86(6): 419-30, 2015 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-26514650

RESUMO

Holstein cattle dominate the global milk production industry because of their outstanding milk production, however, this breed is susceptible to tropical endemic pathogens and suffers from heat stress and thus fewer Holstein populations are raised in tropical areas. The bovine major histocompatibility complex (BoLA)-DRB3 class II gene is used as a marker for disease and immunological traits, and its polymorphism has been studied extensively in Holstein cattle from temperate and cold regions. We studied the genetic diversity of the BoLA-DRB3 gene in South American Holstein populations to determine whether tropical populations have diverged from those bred in temperate and cold regions by selection and/or crossbreeding with local native breeds. We specifically studied Exon 2 of this gene from 855 South American Holstein individuals by a polymerase chain reaction (PCR) sequence-based typing method. We found a high degree of gene diversity at the allelic (Na > 20 and He > 0.87) and molecular (π > 0.080) levels, but a low degree of population structure (FST = 0.009215). A principal components analysis and tree showed that the Bolivian subtropical population had the largest genetic divergence compared with Holsteins bred in temperate or cold regions, and that this population was closely related to Bolivian Creole cattle. Our results suggest that Holstein genetic divergence can be explained by selection and/or gene introgression from local germplasms. This is the first examination of BoLA-DRB3 in Holsteins adapted to tropical environments, and contributes to an ongoing effort to catalog bovine MHC allele frequencies by breed and location.


Assuntos
Bovinos/genética , Genes MHC da Classe II , Antígenos de Histocompatibilidade Classe II/genética , Adaptação Fisiológica , Alelos , Substituição de Aminoácidos , Animais , Cruzamento , Éxons/genética , Variação Genética , Genótipo , Japão , Mutação , Análise de Componente Principal , Seleção Genética , América do Sul , Temperatura , Clima Tropical
2.
Tissue Antigens ; 85(1): 50-7, 2015 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-25430475

RESUMO

Exon 2 of the ovine leukocyte antigen OLA-DRB1 locus was examined in sheep from the Xinjiang Karakul Ram and Bashibai populations, and three generations of hybrids were derived from a cross between Bashibai and Altai Argali wild sheep. This identified 12 novel alleles and 30 previously reported alleles. A neighbor-joining tree of the amino acid sequences of these 42 alleles revealed allelic clusters shared across the study populations. There were significant differences in allelic frequency between Karakul Ram and Bashibai sheep. DRB1*K18cC was the most frequent allele in Kararul Ram with a frequency of 21.2%, while DRB1*2F10c8 (13.2%) and DRB1*0803 (13.2%) were the most frequent alleles found in Bashibai sheep; the alleles DRB1*2F16c2, DRB1*1601, and DRB1*0803 occurred most frequently in F1, F2, and F3 populations, with frequencies of 17.6%, 14.3%, and 20%, respectively. Although many alleles were shared by Bashibai and hybrid sheep, some alleles differed between them, especially in the F1 generation of the Bashibai × Altai Argali cross. The hybrid-specific alleles indicated the introgression of Altai Argali alleles into hybrid flocks. A population tree based on the OLA-DRB1 allelic frequency in each population indicated that the Bashibai sheep and three hybrid populations were similar, with Karakul Ram being genetically distinct.


Assuntos
Alelos , Quimera/genética , Variação Genética , Antígenos de Histocompatibilidade Classe II/genética , Filogenia , Animais , Sequência de Bases , Cruzamento , Quimera/imunologia , China , Cruzamentos Genéticos , Éxons , Feminino , Frequência do Gene , Antígenos de Histocompatibilidade Classe II/classificação , Antígenos de Histocompatibilidade Classe II/imunologia , Masculino , Dados de Sequência Molecular , Ovinos
3.
Tissue Antigens ; 85(1): 35-44, 2015 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-25430590

RESUMO

Bovine leukocyte antigens (BoLAs) are used extensively as markers for bovine disease and immunological traits. In this study, we estimated BoLA-DRB3 allele frequencies using 888 cattle from 10 groups, including seven cattle breeds and three crossbreeds: 99 Red Angus, 100 Black Angus, 81 Chilean Wagyu, 49 Hereford, 95 Hereford × Angus, 71 Hereford × Jersey, 20 Hereford × Overo Colorado, 113 Holstein, 136 Overo Colorado, and 124 Overo Negro cattle. Forty-six BoLA-DRB3 alleles were identified, and each group had between 12 and 29 different BoLA-DRB3 alleles. Overo Negro had the highest number of alleles (29); this breed is considered in Chile to be an 'Old type' European Holstein Friesian descendant. By contrast, we detected 21 alleles in Holstein cattle, which are considered to be a 'Present type' Holstein Friesian cattle. Chilean cattle groups and four Japanese breeds were compared by neighbor-joining trees and a principal component analysis (PCA). The phylogenetic tree showed that Red Angus and Black Angus cattle were in the same clade, crossbreeds were closely related to their parent breeds, and Holstein cattle from Chile were closely related to Holstein cattle in Japan. Overall, the tree provided a thorough description of breed history. It also showed that the Overo Negro breed was closely related to the Holstein breed, consistent with historical data indicating that Overo Negro is an 'Old type' Holstein Friesian cattle. This allelic information will be important for investigating the relationship between major histocompatibility complex (MHC) and disease.


Assuntos
Alelos , Variação Genética , Antígenos de Histocompatibilidade Classe II/genética , Filogenia , Animais , Cruzamento , Bovinos , Chile , Cruzamentos Genéticos , Europa (Continente) , Feminino , Frequência do Gene , Antígenos de Histocompatibilidade Classe II/classificação , Antígenos de Histocompatibilidade Classe II/imunologia , Japão , Masculino , Filogeografia , Análise de Componente Principal
4.
Tissue Antigens ; 83(3): 180-3, 2014 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-24499032

RESUMO

Polymorphisms of the BoLA-DRB3 gene are located primarily in the second exon [antigen binding site (ABS)] and, to a lesser extent, in the upstream regulatory region (URR). It can be hypothesised that exon 2 and the URR are under different types of natural selection. The aim of this work was to determine the URR-exon 2 haplotypes; 34 Holstein samples were genotyped by direct sequencing. A total of 7 URR alleles and 23 exon 2 alleles were detected, and 3 of the URR alleles were novel. Our results may suggest that no relationship exists between the URR and exon 2 of the BoLA-DRB3 gene (linkage disequilibrium P value > 0.05), most likely due to recombination over time. Our results also suggest that both regions of class II genes may be included in the development of new genotyping methods based on next-generation DNA sequencing technologies.


Assuntos
Bovinos/genética , Éxons/genética , Haplótipos/genética , Antígenos de Histocompatibilidade Classe II/genética , Sequências Reguladoras de Ácido Nucleico/genética , Alelos , Animais , Cruzamento , Frequência do Gene/genética , Heterozigoto , Polimorfismo Genético
5.
Meta Gene ; 2: 176-90, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-25606401

RESUMO

Bovine leukocyte antigens (BoLA) are extensively used as markers for bovine disease and immunological traits. However, none of the BoLA genes in Southeast Asian breeds have been characterized by polymerase chain reaction (PCR)-sequence-based typing (SBT). Therefore, we sequenced exon 2 of the BoLA class II DRB3 gene from 1120 individual cows belonging to the Holstein, Sahiwal, Simbrah, Jersey, Brahman, and Philippine native breeds using PCR-SBT. Several cross-breeds were also examined. BoLA-DRB3 PCR-SBT identified 78 previously reported alleles and five novel alleles. The number of BoLA-DRB3 alleles identified in each breed from the Philippines was higher (71 in Philippine native cattle, 58 in Brahman, 46 in Holstein × Sahiwal, and 57 in Philippine native × Brahman) than that identified in breeds from other countries (e.g., 23 alleles in Japanese Black and 35 in Bolivian Yacumeño cattle). A phylogenetic tree based on the DA distance calculated from the BoLA-DRB3 allele frequency showed that Philippine native cattle from different Philippine islands are closely related, and all of them are closely similar to Philippine Brahman cattle but not to native Japanese and Latin American breeds. Furthermore, the BoLA-DRB3 allele frequency in Philippine native cattle from Luzon Island, located in the Northern Philippines was different from that in cattle from Iloilo, Bohol, and Leyte Islands, which are located in the Southern Philippines. Therefore, we conclude that Philippine native cattle can be divided into two populations, North and South areas. Moreover, a neutrality test revealed that Philippine native cattle from Leyte showed significantly greater genetic diversity, which may be maintained by balancing selection. This study shows that Asian breeds have high levels of BoLA-DRB3 polymorphism. This finding, especially the identification of five novel BoLA-DRB3 alleles, will be helpful for future SBT studies of BoLA-DRB3 alleles in East Asian cattle.

6.
Tissue Antigens ; 81(2): 72-82, 2013 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-23216331

RESUMO

Bovine leukemia virus (BLV) is the etiological agent of enzootic bovine leukosis, which is the most common neoplastic disease of cattle. Bovine leukocyte antigen (BoLA) is strongly involved in the subclinical progression of BLV infections. Recent studies show that the BoLA-DRB3 gene might play a direct role in controlling the number of BLV-infected peripheral B lymphocytes in vivo in Holstein cattle. However, the specific BoLA class II allele and DRB3-DQA1 haplotypes determining the BLV proviral load in Japanese Black cattle are yet to be identified. In this study, we focused on the association of BLV proviral load and polymorphism of BoLA class II in Japanese Black cattle. We genotyped 186 BLV-infected, clinically normal cattle for BoLA-DRB3 and BoLA-DQA1 using a polymerase chain reaction-sequence-based typing method. BoLA-DRB3*0902 and BoLA-DRB3*1101 were associated with a low proviral load (LPVL), and BoLA-DRB3*1601 was associated with a high proviral load (HPVL). Furthermore, BoLA-DQA1*0204 and BoLA-DQA1*10012 were related to LPVL and HPVL, respectively. Furthermore, we confirmed the correlation between the DRB3-DQA1 haplotype and BLV proviral load. Two haplotypes, namely 0902B or C (DRB3*0902-DQA1*0204) and 1101A (DRB3*1101-DQA1*10011), were associated with a low BLV proviral load, whereas one haplotype 1601B (DRB3*1601-DQA1*10012) was associated with a high BLV proviral load. We conclude that resistance is a dominant trait and susceptibility is a recessive trait. Additionally, resistant alleles were common between Japanese Black and Holstein cattle, and susceptible alleles differed. This is the first report to identify an association between the DRB3-DQA1 haplotype and variations in BLV proviral load.


Assuntos
Estudos de Associação Genética , Haplótipos/genética , Antígenos de Histocompatibilidade Classe II/genética , Vírus da Leucemia Bovina/imunologia , Leucócitos/imunologia , Provírus/imunologia , Carga Viral/imunologia , Alelos , Animais , Bovinos , Leucose Enzoótica Bovina/genética , Leucose Enzoótica Bovina/imunologia , Leucose Enzoótica Bovina/virologia , Frequência do Gene/genética , Antígenos de Histocompatibilidade Classe II/imunologia , Humanos , Japão
7.
Mol Biol Rep ; 39(7): 7215-20, 2012 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-22531932

RESUMO

Different studies have proved that the resistance/susceptibility to mastitis is genetically determined. The major histocompatibility complex in cows is known as bovine lymphocyte antigen (BoLA). Genes from the BoLA have been associated with the occurrence of infectious diseases such as mastitis and leukosis, especially the BoLA-DRB gene. The object of the present study was to detect associations between BoLA-DRB3 alleles and somatic cell count (SCC), as an indicator of resistance/susceptibility to mastitis in Holstein cattle (N = 123) from La Pampa, Argentina. Fisher's exact test and Woolf-Haldane odds ratio were applied to study the association between SCC and BoLA-DRB3 allele frequencies. Significant association was noted between BoLA-DRB3.2*23 and *27 alleles (p < 0.05) and protective or susceptibility effects, respectively. In addition, alleles BoLA-DRB3.2*20 and *25 exhibit suggestive association with high SCC (p < 0.1). These results were partially in agreement with data reported from Japanese Holstein cattle, but differed from those published by other authors. A possible explanation for the contrasting results could be that the mastitis is a multifactor disease caused by different pathogens. Moreover, most of the studies were carried out using PCR-RFLP method, which has less resolution than PCR-SBT because PCR-RFLP defined alleles included more than one sequenced alleles.


Assuntos
Predisposição Genética para Doença , Antígenos de Histocompatibilidade Classe II/genética , Mastite Bovina/genética , Alelos , Animais , Argentina , Bovinos , Contagem de Células , Feminino , Frequência do Gene , Genótipo , Mastite Bovina/imunologia , Mastite Bovina/microbiologia , Polimorfismo de Nucleotídeo Único
8.
J Dairy Sci ; 95(1): 420-31, 2012 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-22192221

RESUMO

Bovine leukocyte antigen (BoLA), the major histocompatibility complex of cattle, is one of the most polymorphic gene clusters. We genotyped a population of 109 Japanese Black and 39 Holstein cattle to analyze their BoLA class II haplotypes, BoLA-DRB3 locus, 5 BoLA-DQA loci, and 5 BoLA-DQB loci. We identified 26 previously reported DRB3 alleles, 22 previously reported and 3 new DQA alleles, and 24 previously reported and 6 new DQB alleles. A dendrogram was constructed based on the predicted amino acid sequences of the α1 or ß1 domains encoded by BoLA-DQA or -DQB alleles, which revealed that DQA alleles were clustered into 5 loci, whereas DQB alleles could not be clearly assigned to specific DQB loci. The BoLA-DRB3-DQA-DQB haplotypes were sorted by sequential analytical processes, and 42 distinct haplotypes, including 11 previously published haplotypes and 31 novel haplotypes, were defined. Strong linkage disequilibrium was present in the BoLA genes. We also compared DRB3-DQA1 haplotype frequencies between 507 Japanese Black and 143 Holstein cattle. Thirty-nine DRB3-DQA1 haplotypes were identified, including 29 haplotypes from Japanese Black and 22 haplotypes from Holstein cattle. The majority of the haplotypes could be identified in both breeds, although several haplotypes were identified in only a single breed. This is the first report presenting a detailed study of the BoLA class II haplotype in Japanese Black and Holstein cattle in Japan.


Assuntos
Bovinos/genética , Genes MHC da Classe II/genética , Alelos , Sequência de Aminoácidos , Animais , Sequência de Bases , Bovinos/imunologia , Variação Genética/genética , Genótipo , Haplótipos/genética , Dados de Sequência Molecular , Filogenia , Alinhamento de Sequência/veterinária
9.
Tissue Antigens ; 78(3): 208-13, 2011 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-21623735

RESUMO

Recently, two polymerase chain reaction sequence-based typing (PCR-SBT) methods were reported for the genotyping of the bovine leukocyte antigen (BoLA)-DRB3. One technique is a single PCR-SBT (sPCR-SBT) method that generates heterozygous sequences that are subsequently analyzed by the haplofinder program, while the other technique is a nested PCR-SBT (nPCR-SBT) method that allows the analysis of heterozygous sequences using the assign 400ATF software. In this study, these techniques were compared and then integrated to produce an improved genotyping method. The primer set used for sPCR-SBT was more accurate than those used for nPCR-SBT. Combining sPCR-SBT with the assign 400ATF software previously reported for nPCR-SBT enables rapid and accurate genotyping of a large number of DNA samples.


Assuntos
DNA/genética , Antígenos de Histocompatibilidade Classe II/genética , Reação em Cadeia da Polimerase/métodos , Análise de Sequência de DNA , Animais , Sequência de Bases , Bovinos , Primers do DNA/química , Primers do DNA/genética , Genótipo , Heterozigoto , Dados de Sequência Molecular
10.
J Dairy Sci ; 92(6): 2965-70, 2009 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-19448029

RESUMO

Sequence-based typing (SBT) is the most comprehensive method for characterizing major histocompatibility complex (MHC) gene polymorphisms. We report here a new PCR-SBT method for genotyping cattle MHC (BoLA) class II DRB3 using the Assign 400ATF ver. 1.0.2.41 software (Conexio Genomics, Fremantle, Australia), which detects alleles in a semiautomated manner. We examined 12 sets of PCR reactions for their ability to amplify BoLA-DRB3 exon 2 and selected an optimal primer set, which used ERB3N-HL031 for first-round PCR and ALL-DRB3B for second-round PCR. Next, we constructed a BoLA-DRB3 allele database using the reference sequences of the Assign 400ATF software and successfully assigned heterozygous samples (including those with deletion alleles) using bidirectional sequencing, unlike our previously described method, which used unidirectional sequencing for detecting of deletion alleles. Next, blood samples of 128 Holstein cattle were used to correlate the results of our modified PCR-SBT method with those of our previously described PCR-SBT method. Each new PCR-SBT result corresponded completely with the DRB3 allele that was genotyped by our previously described PCR-SBT method. Moreover, we confirmed the accuracy of our modified PCR-SBT method by genotyping 7 sire cattle and their 22 calves using Japanese Black cattle. This new method will contribute to high-throughput genotyping of BoLA-DRB3 by sequence-based typing.


Assuntos
Bovinos/genética , Impressões Digitais de DNA/veterinária , Antígenos de Histocompatibilidade Classe II/genética , Reação em Cadeia da Polimerase/veterinária , Animais , Sequência de Bases , Impressões Digitais de DNA/métodos , Genótipo , Dados de Sequência Molecular , Reação em Cadeia da Polimerase/métodos , Reprodutibilidade dos Testes
11.
Tissue Antigens ; 72(6): 525-31, 2008 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-19000149

RESUMO

Mastitis is an inflammatory response of the mammary gland to irritation, injury, or infectious agents and is a major problem in the dairy industry. We genotyped bovine major histocompatibility complex (BoLA)-DRB3 and BoLA-DQA1 genes in 120 Holstein cattle with clinical mastitis and 85 randomly selected Holstein cattle in Japan by polymerase chain reaction-sequence-based typing. The mastitis cattle were divided into four groups according to the bacterial species that caused the mastitis (Staphylococcus aureus, Streptococci, Escherichia, and coagulase-negative staphylococci). The BoLA-DRB3 and BoLA-DQA1 heterozygosity of each group was compared with that of the control cattle, while the expected heterozygosities based on Hardy-Weinberg proportions and the observed heterozygosities for each locus were compared for each group. The Escherichia-induced and Streptococci-induced mastitis groups showed significant differences between their expected and observed heterozygosities with regard to their BoLA-DQA1 genes. No differences were observed for any group with regard to the BoLA-DRB3 genes. We then found that two BoLA-DQA1 alleles promoted susceptibility to Streptococci-induced mastitis, namely BoLA-DQA1*0101 and BoLA-DQA1*10012 and that the homozygous BoLA-DQA1*0101/0101 and BoLA-DQA1*10011/10011 genotypes promoted susceptibility to mastitis caused by Streptococci and Escherichia, respectively. This is the first report showing that heterozygosity of the BoLA-DQA1 gene is associated with resistance to mastitis progression.


Assuntos
Escherichia , Antígenos de Histocompatibilidade Classe II/genética , Mastite Bovina/genética , Mastite Bovina/microbiologia , Streptococcus , Alelos , Animais , Bovinos , Feminino , Frequência do Gene , Predisposição Genética para Doença , Genótipo , Heterozigoto , Antígenos de Histocompatibilidade Classe II/imunologia , Japão , Mastite Bovina/imunologia
12.
Tissue Antigens ; 72(3): 195-205, 2008 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-18715338

RESUMO

We sequenced the major histocompatibility complex (MHC) class II DQA1 gene in 352 Japanese cattle (95 Japanese Black, 91 Holstein, 102 Japanese Shorthorn and 64 Jersey cattle) using a new sequence-based typing method. In total, 19 bovine MHC (BoLA)-DQA1 alleles, of which two were novel alleles, were detected. The Holstein, Jersey, Japanese Shorthorn and Japanese Black breeds had 13, 12, 10 and 15 alleles, respectively. The dendrogram that was constructed by the neighbor-joining method on the basis of the DQA1 gene allele frequencies of the four Japanese cattle breeds showed that the Holstein and Japanese Black breeds were closest to each other, with Jersey being farther from these two breeds than Japanese Shorthorn. In addition, Wu-Kabat analysis showed that the DQA1 alleles of the Holstein and Japanese Black were the most and least polymorphic, respectively. Phylogenetic analyses indicated that the DQA1 gene of Bovidae such as cattle, sheep, bison and goat were more similar to pig SLA-DQA genes than to human HLA-DQA1 and dog DLA-DQA genes. The cattle, goat, bison, sheep, human and pig DQA1 molecules had similar rates of amino acid sequence polymorphism, but the distribution of their polymorphic residues differed from that in the dog DQA1 protein. However, the Bovidae DQA1 molecule had more polymorphic residues than the human, pig and dog DQA molecules at two regions, namely positions 52-53 and 65-66. This indicates that the Bovidae DQA1 locus is more polymorphic than the DQA loci of other species.


Assuntos
Genes MHC da Classe II , Antígenos de Histocompatibilidade Classe II/genética , Alelos , Animais , Sequência de Bases , Cruzamento , Bovinos , Cães , Variação Genética , Humanos , Japão , Dados de Sequência Molecular , Filogenia , Polimorfismo Genético , Especificidade da Espécie
13.
Kyobu Geka ; 60(13): 1167-70, 2007 Dec.
Artigo em Japonês | MEDLINE | ID: mdl-18078084

RESUMO

A 61-year-old man was pointed out a small peripheral lung nodule and mediastinal lymph node swelling on the chest computed tomography (CT). At the operation, it was diagnosed squamous cell carcinoma and right upper lobectomy and nodal dissection were done. The tumor was 9 mm in size and diagnosed as well differentiated squamous cell carcinoma with metastasis to mediastinal lymph nodes. Postoperative radiotherapy was done (50 Gy). The patient is doing well without apparent recurrence 33 months after surgery. We reported a case of peripheral small squamous cell carcinoma (9 mm) of the lung with metastasis to mediastinal lymph nodes.


Assuntos
Carcinoma de Células Escamosas/patologia , Neoplasias Pulmonares/patologia , Linfonodos/patologia , Metástase Linfática/patologia , Humanos , Masculino , Pessoa de Meia-Idade
14.
Tissue Antigens ; 69(2): 189-99, 2007 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-17257323

RESUMO

In cattle, bovine leukocyte antigens (BoLAs) have been extensively used as markers for bovine diseases and immunological traits. Here, we developed a rapid, high-resolution sequence-based typing (SBT) system for BoLA-DQA1. We amplified 355 bp of BoLA-DQA1 by fully nested polymerase chain reaction (PCR) using the newly constructed primers and then performed direct sequencing of each product. Using this method, we investigated the locus in 51 animals whose BoLA haplotypes had been characterized at the Fifth International BoLA Workshop. We identified 15 distinct DQA1 alleles, and there is no conflict between the typing result of PCR-SBT and restriction fragment length polymorphism analysis. Together with the previously developed method for typing BoLA-DRB3, the PCR-SBT for BoLA-DQA1 clearly provides a useful tool for detailed class IIa haplotype analysis.


Assuntos
Bovinos/imunologia , Antígenos de Histocompatibilidade Classe II/genética , Análise de Sequência de DNA/métodos , Alelos , Animais , Sequência de Bases , Bovinos/genética , Éxons , Haplótipos , Dados de Sequência Molecular , Polimorfismo Genético
15.
Gene ; 316: 111-8, 2003 Oct 16.
Artigo em Inglês | MEDLINE | ID: mdl-14563557

RESUMO

We sequenced exon 2 of the major histocompatibility complex (MHC) class II DRB3 gene from 471 individuals in four different Japanese populations of cattle (201 Japanese Black, 101 Holstein, 100 Japanese Shorthorn, and 69 Jersey cattle) using a new method for sequence-based typing (SBT). We identified the 34 previously reported alleles and four novel alleles. These alleles were 80.0-100.0% identical at the nucleotide level and 77.9-100.0% identical at the amino acid level to the bovine MHC (BoLA)-DRB3 cDNA clone NR1. Among the 38 alleles, eight alleles were found in only one breed in this study. However, these alleles did not form specific clusters on a phylogenetic tree of 236-base pairs (bp) nucleotide sequences. Furthermore, these breeds exhibited similar variations with respect to average frequencies of nucleotides and amino acids, as well as synonymous and non-synonymous substitutions, in all pairwise comparisons of the alleles found in this study. By contrast, analysis of the frequencies of the various BoLA-DRB3 alleles in each breed indicated that DRB3*1101 was the most frequent allele in Holstein cattle (16.8%), DRB3*4501 was the most frequent allele in Jersey cattle (18.1%), DRB3*1201 was the most frequent allele in Japanese Shorthorn cattle (16.0%) and DRB3*1001 was the most frequent allele in Japanese Black cattle (17.4%), indicating that the frequencies of alleles were differed in each breed. In addition, a population tree based on the frequency of BoLA-DRB3 alleles in each breed suggested that Holstein and Japanese Black cattle were the most closely related, and that Jersey cattle were more different from both these breeds than Japanese Shorthorns.


Assuntos
Bovinos/genética , Antígenos de Histocompatibilidade Classe II/genética , Alelos , Sequência de Aminoácidos , Animais , Sequência de Bases , Bovinos/classificação , DNA/química , DNA/genética , Frequência do Gene , Variação Genética , Genótipo , Japão , Dados de Sequência Molecular , Filogenia , Alinhamento de Sequência , Análise de Sequência de DNA , Homologia de Sequência de Aminoácidos , Homologia de Sequência do Ácido Nucleico , Especificidade da Espécie
16.
Eur J Immunogenet ; 30(4): 275-82, 2003 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-12919289

RESUMO

To investigate the genetic diversity of the sheep MHC (Ovar) class II DRB1 locus, we amplified exon 2 of Ovar-DRB1 alleles by polymerase chain reaction (PCR) and determined the nucleotide sequences of both resultant strands after cloning. In our study of a total of 97 sheep of three breeds, namely, Suffolk, Cheviot and Corriedale, we identified 18 previously published alleles and 17 new alleles. These alleles were 83.4 to 94.1% identical at the nucleotide level and 71.4 to 90.9% identical at the amino acid level to Ovar-DRB1*0101. We identified six new alleles in Cheviot sheep and 11 new alleles in Suffolk sheep. Furthermore, we identified 15, 6 and 1 allele in Suffolk, Cheviot and Corriedale sheep, respectively, that have only been found in these breeds to date. Analysis of the frequencies of the various Ovar-DRB1 alleles in each breed indicated that Ovar-DRB1*0702 was the most frequent allele in Suffolk sheep (23.9%), Ovar-DRB1*0203 was the most frequent allele in Cheviot sheep (27.5%) and Ovar-DRB1*0201 was the most frequent allele in Corriedale sheep (25.0%). A comparative analysis of the positions of polymorphic residues in the first extracellular domain of the DRB genes of sheep, humans and mice revealed an extraordinary similarity amongst the positions of polymorphic residues that are associated with the antigen recognition site (ARS). Moreover, the extent of polymorphism seems to be similar in sheep, humans and mice.


Assuntos
Variação Genética , Antígenos de Histocompatibilidade Classe II/genética , Ovinos/genética , Sequência de Aminoácidos , Animais , Humanos , Camundongos , Dados de Sequência Molecular , Estrutura Terciária de Proteína , Alinhamento de Sequência
17.
J Dairy Sci ; 85(6): 1630-2, 2002 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-12146497

RESUMO

A study was made of exon 2 of the bovine leukocyte antigen BoLA-DRB3 gene of 176 Japanese Shorthorn cattle at six farms in Japan using polymerase chain reaction-sequence-based typing (PCR-SBT). An initial round of PCR using conserved locus-specific primers, a second round of PCR using a locus-specific primer, and at least one sequence-specific primer (SSP), followed by direct sequencing of products of PCR with SSP were conducted. Twenty-one BoLA-DRB3 alleles were identified with frequencies ranging from 0.3 to 19.6% in 176 individuals, and two of these alleles were new alleles that have not been reported previously. The three most frequently observed alleles (DRB3*1201, *0301, and *0801) accounted for 43.8% of the alleles in the population of these herds. Next, we tested the products of amplification by PCR of BoLA-DRB3 exon 2 with RsaI, BstYI, and HaeIII, and identified 18 previously described PCR-restriction fragment length polymorphism (RFLP) types. The PCR-RFLP types reflected the results of PCR-SBT exactly. Our results indicate that exon 2 of the BoLA-DRB3 gene is highly polymorphic in Japanese Shorthorn cattle.


Assuntos
Bovinos/genética , Genes MHC da Classe II/imunologia , Complexo Principal de Histocompatibilidade/genética , Polimorfismo de Fragmento de Restrição , Alelos , Sequência de Aminoácidos , Animais , Éxons , Frequência do Gene , Dados de Sequência Molecular , Reação em Cadeia da Polimerase/métodos , Reação em Cadeia da Polimerase/veterinária , Alinhamento de Sequência/veterinária
19.
Ann Thorac Cardiovasc Surg ; 6(4): 236-41, 2000 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-11042479

RESUMO

It is known that prostaglandin E1 (PGE1) is a potent vasodilator and improves red cell deformability. Single lung-ventilation sometimes occurs under lung transplantation, lung cancer surgery and traumatic pneumonectomy, and may result in increased pulmonary resistance, right heart failure and severe hypoxemia. The present experimental study was undertaken to examine the effects of PGE1 on these states induced by single-lung ventilation and hypoventilation. Fourteen pigs weighing 32-33 kg were anesthetized, intubated and ventilated using a respirator and then randomly assigned to two groups, the control group and the PGE1 treated group, 7 pigs each. After median sternotomy to induce severe hypoxemia hypoventilation was induced and then the right hilus pulmonis was cross clamped. Mean blood pressure, mean pulmonary arterial pressure (PAP), pulmonary vessel resistance (PVR), right ventricular stroke work (RVSW) and arterial blood gases (PaO2 and SatO2) were measured at baseline, in the hypoventilation state, and 15 min, 1 hour, and 2 hours after the right hilus pulmonis clamping with hypoventilation. PGE1 (250 microg/20 ml saline) was administered via the central vein starting 15 min after right hilus cross clamping for 1 hour and 45 min in the PGE1 group. PGE1 significantly reduced PAP and PVR, normalized RVSW, and improved PaO2. PGE1 may be useful for the condition of increased pulmonary hypertension during single-lung ventilation and hypoventilation.


Assuntos
Alprostadil/farmacologia , Hipóxia/fisiopatologia , Artéria Pulmonar/fisiologia , Resistência Vascular/efeitos dos fármacos , Vasodilatadores/farmacologia , Função Ventricular Direita/efeitos dos fármacos , Animais , Feminino , Hemodinâmica , Transplante de Pulmão/fisiologia , Masculino , Pneumonectomia , Distribuição Aleatória , Suínos
20.
Surg Today ; 30(9): 785-90, 2000.
Artigo em Inglês | MEDLINE | ID: mdl-11039705

RESUMO

Although the operative mortality following elective aneurysmectomy has achieved satisfactory results, that following surgery for ruptured abdominal aortic aneurysms (AAAs) remains high. The purpose of this study was to identify the factors affecting the mortality rate associated with the treatment of ruptured AAAs. Between 1978 and 1999, 33 patients underwent emergency surgery for a ruptured AAA. The operative mortality was 33.3% and in-hospital mortality was 6.0%. Hypotension, defined as a systolic blood pressure <80 mmHg, was seen in 19 patients at the time of presentation, 9 of whom underwent surgery in this state. In the remaining 10 patients, it was possible to increase the systolic blood pressure to > or =80 mmHg preoperatively. Of the 11 patients who died within 30 days of surgery, 9 had hypotension at the time of induction of anesthesia and only 2 had a systolic blood pressure of > or =80 mmHg. A satisfactory outcome was achieved in patients whose condition met the following criteria: a systolic blood pressure > or =80 mmHg at the time of operation, minimal aortic cross-clamping time, less blood loss and blood transfusions, and a shorter operation time to repair the ruptured AAA. Concomitant heart disease was also found to be an important prognostic factor.


Assuntos
Aneurisma da Aorta Abdominal/cirurgia , Ruptura Aórtica/cirurgia , Adulto , Idoso , Idoso de 80 Anos ou mais , Aneurisma da Aorta Abdominal/mortalidade , Ruptura Aórtica/mortalidade , Perda Sanguínea Cirúrgica , Pressão Sanguínea , Transfusão de Sangue , Emergências , Feminino , Cardiopatias/complicações , Humanos , Hipotensão/complicações , Masculino , Pessoa de Meia-Idade , Cuidados Pré-Operatórios , Prognóstico , Fatores de Risco , Fatores de Tempo , Resultado do Tratamento
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