RESUMEN
OBJECTIVE: To examine factors associated with fertility on dairy farms that used a common fixed-time artificial insemination (FTAI) program in yearling heifers. METHODS: Records were analysed from 954 yearling heifers on 10 south-west Victorian dairy farms that used a common FTAI program, involving the insertion of a 1.9-g progesterone-releasing device for 10 days; 2 mg oestradiol benzoate at insertion; 500 µg cloprostenol on day 7; and FTAI 48 h after device removal. Weight, age, expression of oestrus, sire, semen type (frozen sex-sorted or frozen conventional) and timing of insemination were examined for their relationship with first-service conception rates. RESULTS: Heifers over 300 kg body weight were 1.18-fold more likely to express oestrus during the FTAI program. For every extra 1 kg, there was a 1.5% increase in the likelihood of expressing oestrus. First-service conception rates were 40.3% and 56.0% for sex-sorted and conventional semen, respectively, and were significantly higher when oestrus was expressed. The difference was greater for sex-sorted semen (3.4-fold) compared with conventional semen (1.5-fold). The interval from device removal to insemination varied between 47 and 51.4 h and had no significant effect on conception rates. However, there was a trend towards a higher conception rate for sex-sorted semen when inseminations were performed >50 h after device removal. CONCLUSIONS: Increased fertility was associated with larger heifers and heifers that expressed oestrus, particularly when sexed-sorted semen was used. Variation in the timing of AI with respect to device removal between 47 and 51.4 h did not adversely affect conception rates.
Asunto(s)
Bovinos/fisiología , Fertilidad/fisiología , Inseminación Artificial/veterinaria , Semen/fisiología , Animales , Industria Lechera/métodos , Sincronización del Estro/métodos , Femenino , Fertilización/fisiología , Inseminación Artificial/métodos , EmbarazoRESUMEN
Human follicular B-cell lymphoma is associated with the t(14;18) chromosomal translocation that juxtaposes the Bcl2 proto-oncogene with the immunoglobulin heavy chain (Igh) locus, resulting in the deregulated expression of Bcl2. Our previous studies have shown that the Igh 3' enhancers deregulate the Bcl2 expression in vitro. However, the effects of the Igh 3' enhancer elements on Bcl2 expression in vivo are not known. To investigate the role of the Igh 3' enhancers in Bcl2 deregulation, we used gene targeting to generate knock-in mice in which four DNase I-hypersensitive regions from the murine Igh 3' region were integrated 3' of the Bcl2 locus. Increased levels of Bcl2 mRNA and protein were observed in the B cells of Igh-3'E-bcl2 mice. B cells from Igh-3'E-bcl2 mice showed an extended survival in vitro compared with B cells from wild-type (Wt) mice. The Bcl2 promoter shift from P1 (the 5' promoter) to P2 (the 3' promoter) was observed in B cells from Igh-3'E-bcl2 mice, similar to human t(14;18) lymphomas. The IgH-3'E-bcl2 mice developed monoclonal B-cell follicular lymphomas, which were slowly progressive. These studies show that the Igh 3' enhancers have an important role in the deregulation of Bcl2 and B-cell lymphomagenesis in vivo.
Asunto(s)
Regiones no Traducidas 3'/genética , Linfocitos B/patología , Elementos de Facilitación Genéticos/genética , Genes de las Cadenas Pesadas de las Inmunoglobulinas/genética , Linfoma de Células B/etiología , Proteínas Proto-Oncogénicas/fisiología , Animales , Southern Blotting , Western Blotting , Ciclo Celular , Diferenciación Celular , Supervivencia Celular , Células Cultivadas , Femenino , Citometría de Flujo , Técnica del Anticuerpo Fluorescente , Humanos , Linfoma de Células B/patología , Masculino , Ratones , Ratones Transgénicos , Reacción en Cadena de la Polimerasa , Regiones Promotoras Genéticas/genética , Proto-Oncogenes Mas , Proteínas Proto-Oncogénicas c-bcl-2RESUMEN
Histone deacetylases (HDACs) are frequently overexpressed in broad range of cancer types, where they alter cellular epigenetic programming to promote cell proliferation and survival. However, the mechanism by which HDACs become overexpressed in human cancers remains somewhat of a mystery. In this study, we investigated the expression and functional significance of miR-449a in prostate cancer cells. Using real-time PCR, we found that miR-449a is downregulated in prostate cancer tissues relative to patient-matched control tissue. Introduction of miR-449a into PC-3 prostate cancer cells resulted in cell-cycle arrest, apoptosis and a senescent-like phenotype. In silico analysis of 3'-UTR regions identified a number of genes involved in cell-cycle regulation as putative targets of miR-449a. Using a luciferase 3'-UTR reporter system, we established that HDAC-1 (histone deacetylase 1), a gene that is frequently overexpressed in many types of cancer, is a direct target of miR-449a. Further, our data indicate that miR-449a regulates cell growth and viability in part by repressing the expression of HDAC-1 in prostate cancer cells. Our findings provide new insight into the function of miRNA in regulating HDAC expression in normal versus cancerous tissue.
Asunto(s)
Inhibidores de Histona Desacetilasas , MicroARNs/fisiología , Neoplasias de la Próstata/patología , Línea Celular Tumoral , Proliferación Celular , Inhibidor p27 de las Quinasas Dependientes de la Ciclina , Regulación Enzimológica de la Expresión Génica , Histona Desacetilasa 1 , Humanos , Péptidos y Proteínas de Señalización Intracelular/genética , Masculino , Fenotipo , Neoplasias de la Próstata/enzimología , Neoplasias de la Próstata/genética , ARN Interferente Pequeño/genéticaRESUMEN
Heat shock proteins (HSPs) indicate exposure to cellular stress and adverse cellular effects, thus serving as biomarkers of these effects. The highly conserved Hsp70 proteins are expressed under proteotoxic conditions, whereas small HSPs are expressed in response to stressors acting on the cytoskeleton and cell signaling pathways. Poeciliopsis lucida hepatocellular carcinoma line 1 (PLHC-1) cells have been used extensively for studying effects of cytotoxicity. A number of assays have been developed to examine DNA levels, protein levels, growth rate, morphological changes, and viability. The boundary between sub-lethal and lethal effects of particular stressors has been determined. The methodology and analytical framework for these techniques along with sample assays using cadmium stressed PLHC-1 cells are described. A range of methodologies have been developed in the past decade that allow the analysis and interpretation of gene expression and function in vivo in zebrafish embryos, and many of these are now being applied to the development of embryotoxicity assays. Here we provide the theoretical background and methodology for utilizing Hsp70 expression as an indicator of toxicity in the zebrafish embryo. Hsp70 expression is activated in a tissue-specific manner in zebrafish larvae following exposure to a number of different toxicants, including cadmium. This has allowed the development of an hsp70/eGFP reporter gene system in stable transgenic zebrafish that serves as a reliable yet extremely quick indicator of cell-specific toxicity in the context of the multicellular, living embryo.