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Both epidermal growth factor (EGF) and transforming growth factor (TGF) play an important physiological role in the processes of proliferation and differentiation of several different cell types. However, the expression profiles of these factors in domestic bitches endometrium are still poorly recognized. The aim of the present study was to identify and analyze the differential expression of these factors in various stages of the estrus cycle. Endometrial tissue from proestrus (n=17), estrus (n=10), day 10 diestrus (n=15), day 35 diestrus (n=18) and anestrus (n=25) was collected soon after ovariohysterectomy. Total RNA was isolated from the endometrium by means of Chomczyński and Sacchi method, treated by DNase I, and reverse- transcribed into cDNA. Quantitative analysis of EGF, TGFβ1, TGFβ2, and TGFβ3 cDNA was performed by real-time quantitative polymerase chain reaction (RT-PCR). EGF expression in canine endometrium was increased in the estrus stage as compared to proestrus (P<0.05), day 10 diestrus (P<0.05), day 35 diestrus (P<0.01) and anestrus (P<0.001). We also found the differences in EGF expression between day 10 and day 35 of estrus as well as between day 35 of estrus with anestrus (P<0.05, P<0.01, respectively). The TGFf1 transcript contents were also higher in estrus as compared to other stages (P<0.01). The TGFβ2 and TGFβ3 in the estrus stage was increased compared to proestrus, day 10 diestrus, day 35 diestrus and anestrus (P<0.05). We proved that expression of EGF and TGFβ transcript isoforms is related to the phase of estrus in bitches and therefore may be regulated by specific hormone concentrations during these periods. Our results confirm the hypothesis that these growth factors play a role in the regulation of biochemical changes in the endometrial tissues during the estrus cycle.
Glandular cystic hyperplasia of endometrium/pyometra complex (GCHE-PC) in bitches is a disease which presents several diagnostic and prognostic challenges. The complex pathological process and systemic manifestations of the disease, the potential need for emergency treatment and the owner's desire to maintain the breeding performance of the bitch, are all factors that should be taken into consideration. This review aimed at conducting a thorough diagnostic plan in order to achieve several parameters related to the monitoring and the subsequent treatment of cystic glandular hyperplasia of endometrium pyometra complex. Detailed information on the history and the clinical findings of sick bitches facilitated making an initial selection into two groups: those referred for surgical treatment and those referred for medical treatment. This selection was further clarified by ultrasonographic imaging procedure. USG was found to be an accurate diagnostic tool because of its high level of correlation to patho-anatomical findings. USG facilitates distinguishing densities of the uterine content, internal uterine wall structures and uterine wall thickness.In this study it was found to be a valuable tool in determining the ideal treatment programme and estimating the prognosis for each given case. Ultrasonography proved to be more precise and conclusive than roentgenology in determining the type and location of the pathological process and the extent of the lesion. This review recommends the most potent antibiotics that may be used either concurrently with prostaglandin therapy or in cases when emergency treatment is indicated in bitches with advanced GCHE-PC.
We have studied in the porcine endometrium the expression of oxytocin receptor (OTR) mRNA and the effect of progesterone (P₄) on oxytocin/oxytocin receptor (OT/OTR) function concerning intracellular Ca²⁺mobilisation ([Ca²⁺]i), prostaglandin F2α (PGF2α) and E2 (PGE2; PG) secretion. Tissue was taken from cyclic and early pregnant pigs (days 14-16). A higher expression of OTR mRNA (P<0.05) was observed in the endometrium of cyclic than pregnant pigs. The stimulatory (P<0.05) effect of OT (10⁻⁷ M) on [Ca²⁺]i mobilisation was noticed within 15-60 s and 30-60 s in endometrial stromal cells of cyclic and pregnant pigs, respectively. In the presence of P₄ (10⁻⁵ M) basal and OT-stimulated [Ca²⁺]i concentrations decreased in stromal cells during luteolysis and pregnancy. In stromal cells P₄ delayed mobilisation of [Ca²⁺]i in response to OT by 15 s during luteolysis and had no effect during pregnancy. In cyclic and pregnant epithelial cells OT stimulated mobilisation of [Ca²⁺]i in 45 s and 60 s, respectively. Oxytocin increased (P<0.05) PGF2α secretion during luteolysis and pregnancy and PGE2 during luteolysis from endometrial slices. Progesterone did not inhibit this stimulatory effect. During luteolysis OT increased (P<0.05) PGF2α in epithelial and stromal cells and PGE2 secretion in epithelial cells. In the presence of P₄ this effect of OT was reduced only in stromal cyclic cells (6 h culture). The presence of P₄ decreased the effect of OT on [Ca²⁺]i mobilisation only in stromal cells. We found that, in most conditions, P₄ did not inhibit the OT-stimulated secretion of PG in the porcine endometrium.
The aim of the study was to establish the effect of the ovarian steroids: 17β-oestradiol (E2) and testosterone (T4) on OT-stimulated PGF2α and PGE2 secretion by bovine endometrial cells. The epithelial endometrial cells from days 14-18 of the oestrous cycle (10⁵/ml) were incubated in DMEM/Ham's F12 with 10% FCS (38°C, atmosphere of air and 5% CO₂) for 72-96 h and the last 24 h in DMEM/Ham's F12 with 0.1% BSA. In Exp.l, the doses of steroids used to study their effect on the secretion of PGF2α and PGE2 from endometrial cells stimulated by OT were determined. In Exp.2, the cells were pre-incubated for 30 min with selected doses of steroids: P4 (10⁻⁵ M), T4 (10⁻⁵ M), and E2 (10⁻⁸ M) and next for 4 d with: arachidonic acid (AA; 10⁻⁵ M), OT (10⁻⁷ M) and OT with each of these steroids. The concentration of PGE2 and PGFM -metabolite of PGF2α was determined by EIA. P4, T4, and E2 did not affect (P>0.05) the basal secretion of PGF2α and PGE2, but all the steroids inhibited (P<0.01) OT- stimulated PGF2α secretion. The stimulating effect of OT on PGE2 secretion was not affected by P4 and T4 (P>0.05). This data suggests that different cellular mechanisms exist for steroids affecting the secretion of both prostaglandins from endometrial cells. Moreover, we suggest that non-genomic effect of P4 on bovine endometrial cells is non-specific since the other steroids can impair the effect of OT on these cells. This effect of the steroids can directly modulate function of endometrial cells.
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