RESUMO
The androgen receptor (AR) is a protein encoded by the AR gene, which when mutated may affect spermatogenesis, the process in which spermatozoa are produced; thus, AR mutations could lead to male infertility. We examined exon 1 of the AR gene in men with idiopathic infertility. Blood or semen samples from 111 infertile, oligozoospermic (N = 31), asthenozoospermic (N = 23), teratozoospermic (N = 33), and azoospermic (N = 24) men were analyzed. The extracted DNA was amplified for the exon 1 region of the AR gene. There was a significant correlation between the absence of exon 1 in the AR gene and spermatogenesis defects (P = 0.015). This association was significant in teratozoospermic men (51.5% of the sample). We found that lack of amplification of exon 1 of the AR gene by polymerase chain reaction is associated with morphological defects in the spermogram.
Assuntos
Azoospermia/genética , Éxons , Receptores Androgênicos/genética , Adulto , Brasil , Humanos , MasculinoRESUMO
Gene expression of CDKN1A, CDKN1B, and TP53, and immunostaining of p21, p27 and p53 were evaluated to verify the role of these cell cycle inhibitors in canine prostates with proliferative inflammatory atrophy-PIA and prostatic carcinoma-PC. Seventy samples, 15 normal, 30PIA and 25PC. Regarding number of p27 and p53 labeled cells, difference between normal and PIA and PC was observed, as well as between PIA and PC for p53. Immunostaining intensities of p21, p27 and p53 were different when comparing normal tissues to PIA and PC. Sixteen cDNA of canine prostatic FFPE tissue were subjected to RT-PCR and RT-qPCR, four normal, three PIA, and nine PC. CDKN1A mRNA was detected in four PC by RT-PCR, and it was overexpressed when compared to normal by RT-qPCR, in one PIA and six PC. CDKN1B mRNA was detected in three PC by RT-PCR and it was overexpressed in three PC and decreased in one PC. TP53 mRNA was overexpressed in one PIA and three PC. In conclusion, when overexpressed in canine prostate with premalignant and malignant, p21 and p27 play a role controlling cell proliferation, working as a protective factor in the evolution of PIA to PC, and in the PC development, even in the presence of altered p53.(AU)
A expressão gênica de CDKN1A, CDKN1B e TP53, assim como imunomarcação de p21, p27 e p53 foram realizadas a fim de verificar o papel desses inibidores do ciclo celular na próstata canina com atrofia inflamatória proliferativa (PIA) e carcinoma prostático (PC). Foram obtidas70 amostras de próstata canina, sendo 15 de tecido normal, 30 de PIA e 25 de PC. Quanto ao número de células imunomarcadas foi observada diferença entre amostras normais, com PIA e PC para p27 e p53, assim como entre PIA e PC para p53. Para a intensidade de imunomarcação houve diferença entre os tecidos normais e com PIA e PC para p21, p27 e p53. Foram obtidas dezesseis amostras de cDNA a partir de amostras de próstatas caninas embebidas em parafina para a realização da RT-PCR e RT-qPCR, sendo quatro normais, três com PIA, e nove com o PC. O gene CDKN1A foi detectado em quatro das amostras com PC por RT-PCR, e pela RT-qPCR este estava superexpresso em uma PIA e em seis PC quando da comparação com o tecido prostático normal. O CDKN1B foi detectado em três PC por RT-PCR e pela RT-qPCR estava superexpresso em três PC e reduzido em um PC. O TP53 foi detectado em todas as próstatas caninas com PIA e PC por RT-PCR, sendo também superexpresso em uma glândula com PIA e em três com PC. Concluiu-se que p21 e p27 quando superexpressas na próstata canina com lesões pré-malignas (PIA) e malignas (PC) desempenham ação no controle da proliferação celular, possivelmente atuando como fator de proteção na evolução da PIA para PC, e no desenvolvimento do PC, mesmo na presença de p53 alterada. Assim, o próximo passo é avaliar essas proteínas do ciclo celular em casos de PC canino com metástase.(AU)
Assuntos
Animais , Cães , Próstata/fisiologia , Atrofia/diagnóstico , Carcinoma , Ciclo Celular , Cães/anatomia & histologia , Cães/anormalidadesRESUMO
Paracoccidioides brasiliensis is a pathogenic fungus that undergoes a temperature-dependent cell morphology change from mycelium (22 degrees C) to yeast (36 degrees C). It is assumed that this morphological transition correlates with the infection of the human host. Our goal was to identify genes expressed in the mycelium (M) and yeast (Y) forms by EST sequencing in order to generate a partial map of the fungus transcriptome. Individual EST sequences were clustered by the CAP3 program and annotated using Blastx similarity analysis and InterPro Scan. Three different databases, GenBank nr, COG (clusters of orthologous groups) and GO (gene ontology) were used for annotation. A total of 3,938 (Y = 1,654 and M = 2,274) ESTs were sequenced and clustered into 597 contigs and 1,563 singlets, making up a total of 2,160 genes, which possibly represent one-quarter of the complete gene repertoire in P. brasiliensis. From this total, 1,040 were successfully annotated and 894 could be classified in 18 functional COG categories as follows: cellular metabolism (44%); information storage and processing (25%); cellular processes-cell division, posttranslational modifications, among others (19%); and genes of unknown functions (12%). Computer analysis enabled us to identify some genes potentially involved in the dimorphic transition and drug resistance. Furthermore, computer subtraction analysis revealed several genes possibly expressed in stage-specific forms of P. brasiliensis. Further analysis of these genes may provide new insights into the pathology and differentiation of P. brasiliensis.