RESUMO
MicroRNAs (miRNAs) are small non-coding RNAs that participate in the regulation of gene expression. Their role during mammary gland development is still largely unknown. In this study, we performed a microarray analysis to identify miRNAs associated with high mammogenic potential of the bovine mammary gland. We identified 54 significantly differentially expressed miRNAs between the mammary tissue of dairy (Holstein-Friesian, HF) and beef (Limousin, LM) postpubertal heifers. Fifty-two miRNAs had higher expression in the mammary tissue of LM heifers. The expression of the top candidate miRNAs (bta-miR-10b, bta-miR-29b, bta-miR-101, bta-miR-375, bta-miR-2285t, bta-miR-146b, bta-let7b, bta-miR-107, bta-miR-1434-3p) identified in the microarray experiment was additionally evaluated by qPCR. Enrichment analyses for targeted genes revealed that the major differences between miRNA expression in the mammary gland of HF versus LM were associated with the regulation of signalling pathways that are crucial for mammary gland development, such as TGF-beta, insulin, WNT and inflammatory pathways. Moreover, a number of genes potentially targeted by significantly differentially expressed miRNAs were associated with the activity of mammary stem cells. These data indicate that the high developmental potential of the mammary gland in dairy cattle, leading to high milk productivity, depends also on a specific miRNA expression pattern.
Assuntos
Bovinos/genética , Glândulas Mamárias Animais/química , MicroRNAs/análise , Animais , Bovinos/classificação , Indústria de Laticínios , Feminino , Glândulas Mamárias Animais/crescimento & desenvolvimento , MicroRNAs/genética , Reação em Cadeia da Polimerase , Carne Vermelha , Transdução de Sinais , Nicho de Células-Tronco , Células-Tronco/metabolismoRESUMO
The aim of the study was to estimate the size of bone marrow-origin stem/progenitor population in 2-year old nonpregnant Holstein-Friesian heifers. Quantitative and qualitative analysis was done using scanning cytometry and confocal microscopy of mammary tissue slices labelled with the combination of two markers: Sca-1 (marker of stem-progenitor cells) and CD45 (marker of hematopoietic cells). The average (+/- SEM) percentage of Sca-1POS CD45 POS cells was 0.89 +/- 0.21. They were localized mainly outside of mammary ducts, in the stroma and sometimes intraluminally. Our results indicate that the subpopulation of Sca-1POS cells bearing CD45 antigen may enrich the niche of mammary stem/progenitor cells from the bone marrow and participate in the growth of the mammary gland in post-pubertal heifers.
Assuntos
Células da Medula Óssea/citologia , Bovinos/fisiologia , Glândulas Mamárias Animais/citologia , Células-Tronco/citologia , Animais , Feminino , Regulação da Expressão Gênica/fisiologia , Citometria de Varredura a Laser , Microscopia ConfocalRESUMO
Equid herpesvirus type 2 (EHV-2) together with equid herpesvirus type 5 are members of Gammaherpesvirinae subfamily, genus Rhadinovirus. EHV-2 is one of major agents causing diseases of horses common worldwide. A possible role of EHV-2 in reactivating latent equid herpesvirus type-1 has been suggested, because reactivation of latent EHV-1 was always accompanied by EHV-2 replication. Variety techniques, including cell culture, PCR and its modifications, have been used to diagnose EHV-2 infections. The aim of this study was to develop, optimize and determine specificity of real-time PCR (qPCR) for EHV-2 DNA detection using HybProbes chemistry and to evaluate clinical samples with this method. The analytical sensitivity of assay was tested using serial dilutions of viral DNA in range between 70 and 7x10(5) copies/ml. The limit of detection (LOD) was calculated using probit analysis and was determined as 56 copies/ml. In further studies 20 different clinical samples were tested for the presence of EHV-2. Described in-house qPCR method detected viral DNA in 5 of 20 specimens used. The results of this work show that developed HybProbes-based real-time PCR assay is very reliable and valuable for detection and quantification of equid herpesvirus type 2 DNA in different clinical samples. The high level of sensitivity, accuracy and rapidity provided by the LightCycler 2.0 instrument are favorable for the use of this system in the detection of EHV-2 DNA in veterinary
Assuntos
DNA Viral/genética , DNA Viral/isolamento & purificação , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Rhadinovirus/genética , Rhadinovirus/isolamento & purificação , Animais , Infecções por Herpesviridae/diagnóstico , Infecções por Herpesviridae/veterinária , Infecções por Herpesviridae/virologia , Doenças dos Cavalos/diagnóstico , Doenças dos Cavalos/virologia , Cavalos , Reação em Cadeia da Polimerase em Tempo Real/métodos , Sensibilidade e Especificidade , Infecções Tumorais por Vírus/diagnóstico , Infecções Tumorais por Vírus/veterinária , Infecções Tumorais por Vírus/virologiaRESUMO
We have analysed all the reported and diagnosed cases of scabies in the years 1999-2000 per 100 000 inhabitants in the Lódz voivodeship. The data was obtained from the Regional Administration Unit for the Control of Epidemics and Hygiene Promotion in Lódz. The incidence in Lódz voivodeship was found to be twice as high (89,0) as that in Poland (44,0). The highest incidence in Lódz voivodeship was found in Radomsko (268,0), Pabianice (149,0) and Kutno (148.5) districts. The number of new cases registered in 1999-2000 did not increase in the districts: Leczyca, Radomsko, Lowicz and Belchatów.
