An approach to uncover the relationship between 17b-estradiol and ESR1/ESR2 ratio in the regulation of canine corpus luteum.

The canine corpus luteum (CL) is able to synthetise, activate and deactivate 17b-estradiol (E2) and also expresses nuclear estrogen receptors in a time-dependent manner during diestrus. Nevertheless, we are still missing a better comprehension of E2 functions in the canine CL, especially regarding the specific roles of estrogen receptor alpha (ERa) and ERb, encoded by ESR1 and 2, respectively. For that purpose, we analyzed transcriptomic data of canine non-pregnant CL collected on days 10, 20, 30, 40, 50 and 60 of diestrus and searched for differentially expressed genes (DEG) containing predicted transcription factor binding sites (TFBS) for ESR1 or ESR2. Based on biological functions of DEG presenting TFBS, expression of select transcripts and corresponding proteins was assessed. Additionally, luteal cells were collected across specific time points during diestrus and specificity of E2 responses was tested using ERa and/or ERb inhibitors. Bioinformatic analyses revealed 517 DEGs containing TFBS, from which 67 for both receptors. In general, abundance of predicted ESR1 targets was greater in the beginning, while abundance of ESR2 targets was greater in the end of diestrus. ESR1/ESR2 ratio shifted from an increasing to a decreasing pattern from day 30 to 40 post ovulation. Specific receptor inhibition suggested an ERa-mediated positive regulation of CL function at the beginning of diestrus and an ERb-mediated effect contributing to luteal regression. In conclusion, our data points toward a broad spectrum of action of E2 and its nuclear receptors, which can also act as transcription factors for other genes regulating canine CL function.

Is the canine corpus luteum an insulin-sensitive tissue?

This study aimed to determine in the canine corpus luteum throughout the dioestrus (1) the influence of insulin on glucose uptake; (2) the regulation of genes potentially involved; and (3) the influence of hypoxia on glucose transporter expression and steroidogenesis, after treatment with cobalt chloride (CoCl2). Glucose uptake by luteal cells increased 2.7 folds (P < 0.05) in response to insulin; a phenomenon related to increased expression of glucose transporter (GLUT) 4 and phosphorylation of protein kinase B (AKT). The gene expression of insulin receptor and SLC2A4 (codifier of GLUT4) genes after insulin stimulation increased on day 20 post ovulation (p.o.) and declined on day 40 p.o. (P < 0.05). Regarding potentially involved molecular mechanisms, the nuclear factor kappa B gene RELA was upregulated on days 30/40 p.o., when SLC2A4 mRNA was low, and the interleukin 6 (IL6) gene was upregulated in the first half of dioestrus, when SLC2A4 mRNA was high. CoCl2 in luteal cell cultures increased the hypoxia-inducible factor HIF1A/HIF1A and the SLC2A4/GLUT4 expression, and decreased progesterone (P4) production and hydroxyl-delta-5-steroid dehydrogenase 3 beta (HSD3B) mRNA expression (P < 0.05). This study shows that the canine luteal cells are responsive to insulin, which stimulates glucose uptake in AKT/GLUT4-mediated pathway; that may be related to local activity of RELA and IL6. Besides, the study reveals that luteal cells under hypoxia activate HIF1A-modulating luteal function and insulin-stimulated glucose uptake. These data indicate that insulin regulates luteal cells' glucose disposal, participating in the maintenance and functionality of the corpus luteum.

Development and morphological changes in the vaginal closure membrane throughout gestation in Galea spixii (Rodentia: Caviidae).

Present research was carried out in order to perform the monitoring of development, recognizes the type of tissue and describes histological and cellular changes of the vaginal closure membrane (VCM) throughout pregnancy in Galea spixii. The results showed that at 20 days of gestation (DG), the VCM occludes completely the external vaginal ostium. Microscopically, the VCM presented juxtaposed cells, derived from the stratum germinative of the stratified epithelium of vaginal mucosa at 20 DG and areas with cell clusters with the presence of intercellular spaces in the final stages of pregnancy (40-50 DG). At 0 DG, the stratified epithelium of vaginal mucosa presented all strata but at 20 DG presented stratified epithelium without the stratum corneum and stratum granular and showed communicant junctions by desmosomes and interdigitations in the cell membrane compound the VCM. Gradually from 40 to 50 DG the stratum germinative became barely perceptible. Many cells showed apoptotic nuclei and emerged many intercellular spacing. So, the interdigitations and desmosomes were not observed. Here, it was demonstrated for the first time that the VCM is formed after the extinction of the stratum granular and corneum of the vaginal mucosa epithelium, with the proliferation of the cells of stratum germinative and communication and junction through desmosomes and interdigitations of these cells. At the end of pregnancy, cellular apoptosis; the spread of stratum germinative; and, absence of cellular communication and junction may be responsible for the weakening of the VCM and may assist the process of rupture of this membrane.

Glucose transporter 1 expression accompanies hypoxia sensing in the cyclic canine corpus luteum.

The canine corpus luteum (CL) functions as a source of progesterone (P4) and 17ß-oestradiol (E2); however, the transport of energy substrates to maintain its high hormonal output has not yet been characterised. This study involved the localisation and temporal distribution of the facilitative glucose transporter 1 and the quantification of the corresponding protein (GLUT1) and gene (SLC2A1) expression. Some GLUT1/SLC2A1 regulatory proteins, such as hypoxia-inducible factor 1α (HIF1A) and fibroblast growth factor 2 (FGF2); mRNAs, such as HIF1A, FGF2 and vascular endothelial growth factor A (VEGFA); and VEGFA receptors 1 and 2 (FLT1 and KDR) were also analysed from days 10 to 70 after ovulation. Additionally, plasma P4 and E2 levels were assessed via chemiluminescence. Moreover, the canine KDR sequence has been cloned, thereby enabling subsequent semi-quantitative PCR analysis. Our results demonstrate time-dependent variations in the expression profile of SLC2A1 during dioestrus, which were accompanied by highly correlated changes (0.84

Características e evidências da produção científica de enfermeiros sobre erros de medicação no ambiente hospitalar / Characteristics and evidence of nursing scientific production for medication errors at the hospital environment

Estudo que objetivou identificar as características das publicações de enfermeiros sobre erros de medicação. Metodologia de revisão integrativa abrangendo janeiro de 2005 a outubro de 2010 com os descritores “erros de medicação” e “enfermagem” com dados coletados em bases eletrônicas através do Portal Capes. Resultados mostram quatro categorias: condutas dos profissionais diante do erro de medicação; tipos e taxas de erros; fragilidades no sistema de medicação e barreiras ao erro. A conduta prevalente foi não notificar o erro; o tipo prevalente de erro foi na administração e as taxas de erro oscilaram entre 14,8 a 56,7%. A caligrafia ilegível, falhas de comunicação entre profissionais e falta de conhecimento técnico foram fragilidades apontadas. Entre barreiras teve-se a educação do paciente, da enfermagem e a tecnologia. Constataram-se avanços nas pesquisas que testam barreiras e evidenciaram-se lacunas na falta de estudos que abordem aspectos farmacodinâmicos ou farmacocinéticos dos medicamentos envolvidos em erros.