RESUMO
Seventeen commercial and research laboratories participated in two comparison tests under the auspices of the International Society for Animal Genetics to develop an internationally tested, microsatellite-based parentage and identification panel for the domestic cat (Felis catus). Genetic marker selection was based on the polymorphism information content and allele ranges from seven random-bred populations (n = 261) from the USA, Europe and Brazil and eight breeds (n = 200) from the USA. Nineteen microsatellite markers were included in the comparison test and genotyped across the samples. Based on robustness and efficiency, nine autosomal microsatellite markers were ultimately selected as a single multiplex 'core' panel for cat identification and parentage testing. Most markers contained dinucleotide repeats. In addition to the autosomal markers, the panel included two gender-specific markers, amelogenin and zinc-finger XY, which produced genotypes for both the X and Y chromosomes. This international cat parentage and identification panel has a power of exclusion comparable to panels used in other species, ranging from 90.08% to 99.79% across breeds and 99.47% to 99.87% in random-bred cat populations.
Assuntos
Gatos/classificação , Repetições de Microssatélites , Alelos , Animais , Gatos/genética , Marcadores Genéticos , Genótipo , Polimorfismo GenéticoRESUMO
Several specific cytogenetic changes are known to be associated with childhood acute lymphoblastic leukemia (ALL), and many of them are important prognostic factors for the disease. Little is known, however, about the changes in gene expression in ALL. Recently, the development of cDNA array technology has enabled the study of expression of hundreds to thousands of genes in a single experiment. We used the cDNA array method to study the gene expression profiles of 17 children with precursor-B ALL. Normal B cells from adenoids were used as reference material. We discuss the 25 genes that were most over-expressed compared to the reference. These included four genes that are normally expressed only in the myeloid lineages of the hematopoietic cells: RNASE2, GCSFR, PRTN3 and CLC. We also detected over-expression of S100A12, expressed in nerve cells but also in myeloid cells. In addition to the myeloid-specific genes, other over-expressed genes included AML1, LCP2 and FGF6. In conclusion, our study revealed novel information about gene expression in childhood ALL. The data obtained may contribute to further studies of the pathogenesis and prognosis of childhood ALL.
Assuntos
Antígenos de Neoplasias/genética , Biomarcadores Tumorais/metabolismo , DNA de Neoplasias/análise , Genes Neoplásicos/genética , Células Mieloides/metabolismo , Leucemia-Linfoma Linfoblástico de Células Precursoras/genética , Doença Aguda , Adolescente , Criança , Pré-Escolar , Primers do DNA/química , Feminino , Perfilação da Expressão Gênica , Humanos , Lactente , Cariotipagem , Masculino , Células Mieloides/patologia , Análise de Sequência com Séries de Oligonucleotídeos , Leucemia-Linfoma Linfoblástico de Células Precursoras/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
The molecular and genetic characteristics of ameloblastoma are still poorly understood. We analyzed gene expression in fresh-frozen ameloblastomas and human fetal tooth germs, using a cDNA microarray. Thirty-four genes exhibited significant changes in expression levels in the ameloblastoma. Eleven genes were overexpressed more than three-fold, and 23 genes were underexpressed to below 0.4 of the control level. The oncogene FOS was the most overexpressed gene (from eight- to 14-fold), followed by tumor-necrosis-factor-receptor 1 (TNFRSF1A). Genes for sonic hedgehog (SHH), TNF-receptor-associated-factor 3 (TRAF3), rhoGTP-ase-activating protein 4 (ARHGAP4), deleted in colorectal carcinoma (DCC), cadherins 12 and 13 (CDH12 and 13), teratocarcinoma-derived growth-factor-1 (TDGF1), and transforming growth-factor-beta1 (TGFB1) were underexpressed in all tumors. In selected genes, a comparison between cDNA microarray and real-time RT-PCR confirmed similar relative gene expression changes. The gene expression profile identifies candidate genes that may be involved in the origination of ameloblastoma and several genes previously unidentified in relation to human tooth development.
Assuntos
Ameloblastoma/genética , Fator de Crescimento Epidérmico , Perfilação da Expressão Gênica/métodos , Análise de Sequência com Séries de Oligonucleotídeos/métodos , Germe de Dente/metabolismo , Adolescente , Adulto , Idoso , Antígenos CD/genética , Caderinas/genética , Indução Embrionária/genética , Feminino , Proteínas Ligadas por GPI , Proteínas Ativadoras de GTPase/genética , Genes fos/genética , Substâncias de Crescimento/genética , Proteínas Hedgehog , Humanos , Peptídeos e Proteínas de Sinalização Intercelular , Análise dos Mínimos Quadrados , Masculino , Glicoproteínas de Membrana/genética , Pessoa de Meia-Idade , Proteínas de Neoplasias/genética , Odontogênese/genética , Proteínas/genética , Receptores do Fator de Necrose Tumoral/genética , Receptores Tipo I de Fatores de Necrose Tumoral , Análise de Regressão , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fator 3 Associado a Receptor de TNF , Germe de Dente/embriologia , Transativadores/genética , Fator de Crescimento Transformador beta/genética , Fator de Crescimento Transformador beta1 , Dedos de Zinco/genéticaRESUMO
BACKGROUND: Genetic aberrations provide prognostic information in childhood ALL. The proportion of patients with detectable aberrations can be increased by combining G-banding with comparative genomic hybridization (CGH). PROCEDURE: We studied 79 children with ALL by CGH and G-banding, and explored the relationship of these findings to clinical features and outcome. RESULTS: CGH revealed DNA copy number changes in 57 patients (72%), 9 of whom had normal karyotype by G-banding. Gains were more frequent than losses, and changes of whole chromosomes more frequent than partial aberrations. Two frequent partial losses were found; at 9p and 12p. The 9 patients with loss at 12p were studied for the deletion of TEL (ETV6) gene and the fusion of TEL and AML1 genes by fluorescent in situ hybridization (FISH). Eight out of the 9 children with loss at 12p harbored the TEL-AML1 translocation and all 9 had the deletion of a nontranslocated TEL allele. All 9 had precursor-B phenotype and L1 morphology, and 8/9 had WBC below 50 x 10(9)/liter. All children were treated according to Nordic ALL protocols, had a good response to treatment based on day 15 bone marrow morphology, and 7 out of the 9 survived in continuous complete remission (median follow-up 74 months). CONCLUSIONS: CGH is a valuable tool in screening for genetic aberrations in childhood ALL. DNA copy number losses detected at 12p associate with TEL-AML1 fusion as well as with favorable prognostic features.
Assuntos
Cromossomos Humanos Par 12/genética , Cromossomos Humanos Par 9/genética , DNA de Neoplasias/genética , Proteínas de Fusão Oncogênica/genética , Leucemia-Linfoma Linfoblástico de Células Precursoras/genética , Adolescente , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Criança , Pré-Escolar , Subunidade alfa 2 de Fator de Ligação ao Core , Feminino , Humanos , Lactente , Cariotipagem , Perda de Heterozigosidade , Masculino , Hibridização de Ácido Nucleico , Fenótipo , Leucemia-Linfoma Linfoblástico de Células Precursoras/tratamento farmacológico , Leucemia-Linfoma Linfoblástico de Células Precursoras/patologia , Prognóstico , Análise de SobrevidaAssuntos
Proteínas do Esmalte Dentário/genética , Genes sry , Doenças dos Cavalos/genética , Aberrações dos Cromossomos Sexuais/veterinária , Cromossomo X , Amelogenina , Animais , Diagnóstico Diferencial , Feminino , Doenças dos Cavalos/diagnóstico , Cavalos , Hibridização in Situ Fluorescente/veterinária , Cariotipagem/veterinária , Reação em Cadeia da Polimerase/veterináriaRESUMO
BACKGROUND AND OBJECTIVE: We previously found a high-level amplification in chromosomal region 21q22 in two children with acute lymphoblastic leukemia (ALL) using comparative genomic hybridization. The same region harbors the AML1 gene. The aim of the present study was to investigate whether AML1 is a target gene in these amplifications. DESIGN AND METHODS: Bone marrow samples were obtained from 112 childhood ALL patients. The copy number of AML1 was studied using fluorescent in situ hybridization with a dual color DNA probe specific for the AML1 and TEL genes. RESULTS: Three of the patients had 3-to-8 fold amplification of AML1 and showed a high-level amplification of 21q22 by comparative genomic hybridization. In two of them the extra copies were shown to be located tandemly in a derivative of chromosome 21. Thirty-seven of the patients (33%) had 1-to-2 extra copies of AML1, most probably reflecting the incidence of trisomy 21 and tetrasomy 21. The TEL-AML1 fusion was less frequent in the patients with extra copies of AML1 (7/40; 18%) than in the patients with no extra copy (24/72; 33%). None of the three patients with 3-to-8 fold amplification of AML1 showed the fusion or loss of TEL. INTERPRETATION AND CONCLUSIONS: Our findings suggest that the AML1 gene is a target gene in the 21q22 amplicon in childhood ALL. To understand the role, if any, of the AML1 amplification in leukemogenesis, further studies are needed.
Assuntos
Proteínas de Ligação a DNA/genética , Leucemia-Linfoma Linfoblástico de Células Precursoras/genética , Proteínas Proto-Oncogênicas , Fatores de Transcrição/genética , Adolescente , Células da Medula Óssea/metabolismo , Criança , Pré-Escolar , Cromossomos Humanos Par 21 , Subunidade alfa 2 de Fator de Ligação ao Core , Feminino , Amplificação de Genes/genética , Dosagem de Genes , Humanos , Hibridização in Situ Fluorescente , Lactente , Masculino , Proteínas de Neoplasias/genéticaRESUMO
Phenotypes of cat plasma apolipoprotein A4 (APOA4), antithrombin 3 (AT3), alpha 1B-glycoprotein (A1BG), transferrin (TF), vitamin D-binding protein (GC), and an unidentified pretransferrin (PTF) were determined by using simple methods of horizontal, nondenaturing gel electrophoresis followed by protein staining. The cat proteins were identified by immunoblotting using antisera for human plasma proteins. Three alleles were reported for each of TF and PTF, and two alleles were reported for each of GC, APOA4, AT3, and A1BG. The mongrels and Persians showed a high degree of polymorphism at most of the loci whereas the Birmans exhibited much less variation. Genetic evidence indicating the occurrence of a monomeric and a dimeric form of APOA4 in cat plasma was reported.
Assuntos
Proteínas Sanguíneas/genética , Gatos/genética , Polimorfismo Genético , Animais , Antitrombina III/genética , Apolipoproteínas A/genética , Eletroforese em Gel Bidimensional , Eletroforese em Gel de Poliacrilamida , Frequência do Gene , Immunoblotting , Precursores de Proteínas/genética , Transferrina/genética , Proteína de Ligação a Vitamina D/genéticaRESUMO
Plasma samples of 270 foxes from 45 complete families (14 of arctic foxes, 28 of silver foxes and 3 with arctic x silver fox hybrid offspring) were analysed by a method of two-dimensional agarose gel (pH 5.4)-horizontal polyacrylamide gel (pH 9.0) electrophoresis followed by general-protein staining of gels. Genetic polymorphism of three plasma proteins, tentatively designated prealbumin (Pr), postalbumin 1 (Pa1), and pretransferin 1 (Prt1), was observed. In silver foxes, Pa1 and Prt1 showed a high degree of polymorphism, each with 3 common alleles, while Pr showed a scarce polymorphism. The arctic foxes were monomorphic for Pr and Prt1 and showed a scarce Pa1 polymorphism. The Prt1 phenotype of arctic foxes showed identical mobility with one of the Prt1 variants of silver foxes.
Assuntos
Eletroforese das Proteínas Sanguíneas/métodos , Proteínas Sanguíneas/genética , Raposas/genética , Animais , Animais Domésticos/genética , Cruzamentos Genéticos , Eletroforese em Gel de Ágar , Eletroforese em Gel de Poliacrilamida , Raposas/sangue , Marcadores Genéticos , Polimorfismo Genético , Especificidade da EspécieRESUMO
Plasma samples of 235 foxes from 38 complete families (14 of arctic foxes, 21 of silver foxes and 3 with arctic x silver fox hybrid offspring) were analysed by one-dimensional horizontal polyacrylamide gel electrophoresis (PAGE) pH 9.0 followed by general-protein staining of gels. A major postalbumin of fox plasma was identified as alpha 1B-glycoprotein (alpha 1B) by using immunoblotting with antiser m specific to human or pig plasma alpha 1B. Four codominant, autosomal alleles of alpha 1B were found in arctic foxes. Two transferrin (TF) alleles (TfF, TfS) were observed in arctic foxes and two (TfD, Tff) in silver foxes; the TF F type of both of the fox species showed identical electrophoretic mobilities. The arctic foxes showed a high degree of polymorphism for both TF and alpha 1B. The silver foxes showed a scarce polymorphism of TF and were monomorphic for alpha 1B. The arctic fox, silver fox and their hybrids could be clearly differentiated from one another by their plasma protein patterns obtained by the PAGE method.