Influence of genistein and diadizine on regularity of estrous cycle in cyclic female Wistar rat: interaction with estradiol receptors and vascular endothelial growth factor.

Background: Isoflavones are estrogenic compounds that exist in soy, clover, and peanuts. They are selective estrogen receptor modulators. Aim: The study was planned to explain the interactions of isoflavones with estrogen receptors alpha (ERα), beta (ERß), and vascular endothelial growth factor (VEGF) expressions in ovarian and uterine tissues during different stages of the estrous cycle of regular cyclic female Wistar rats. Methods: Thirty-two regular cyclic females were divided equally into control group: fed casein-based diet and isoflavones group: fed casein-based diet and gavaged 50 mg/kg/day soy isoflavones extract 40%. The regularity of estrus cycles was monitored. Final body weight (FBW), weight gain (BWG), and ovarian and uterine weights were estimated. Histopathology and immunohistochemistry for ERα, Erß, and VEGF in ovarian and uterine tissues were performed. Results: All females (100%, n = 16) in control group showed regularity in estrous cycle compared to 62.5% (n = 10) in isoflavones group. Estrus and diestrus phases revealed prolongation and shortening in isoflavones rats than control, respectively. Nonsignificant variation was noted in the duration of the whole cycle of both groups. FBW and BWG significantly decreased however, ovarian and uterine weights increased significantly in all estrous phases of isoflavones group than control. Histopathology demonstrated an increase in number of follicles/ovaries besides, hyperplasia and proliferation of luminal epithelium with hydropic degeneration in the isoflavones group. Also, uterine connective tissue stroma showed edema in the isoflavones group during all estrous phases. Immunostaining percentages of ERα, Erß, and VEGF protein expression were significantly elevated in the isoflavones group during all estrous phases. Conclusion: Isoflavones induced irregularity of the estrous cycle that was encountered by increased and altered ERα, Erß, and VEGF expressions in ovarian and uterine tissues.

Subclinical endometritis and postpartum ovarian resumption in respect to TNF-α, IL-8 and CRP in Egyptian buffaloes.

The study was carried out to find the relation between subclinical endometritis (SCE) and postpartum (pp.) ovarian resumption as well as to evaluate serum and endometrial TNF-α, IL-8 and serum CRP in buffaloes with and without SCE. Thirty-nine pluriparous buffaloes at the 3rd (W3), 5th (W5) and 7th (W7) week pp. were involved in this experiment. The parity of the buffaloes ranged from 4 to 8 with an average 5.8±0.2. Subclinical endometritis was diagnosed by the percentage of polymorphonuclear cells (PMNs) in uterine cytology obtained from endometrial cytobrush at W5 and W7. The cut-off point of PMNs% in buffaloes for SCE was ≥ 6% at W5 or ≥ 4% at W7. According to PMNs%, buffaloes were divided into SCE group (n=27) and non-SCE group (n=12). Ovarian cyclicity was monitored by rectal palpation, ultrasonography and progesterone assay at W3, W5 and W7. Serum and endometrial TNF-α, IL-8 and serum CRP were estimated at W5 and W7. Buffaloes with SCE (55.6%) showed delayed ovarian activity as compared to non-SCE (16.7%) animals (P=0.036). Significant increase in serum cytokines and CRP levels were detected at W5 (P ˂0.05) and W7 (P <0.01) in SCE buffaloes as compared to non-SCE. Endometrial levels of cytokines were significantly (P ˂0.05) elevated in SCE buffaloes. Serum and endometrial cytokines showed significant positive correlation. Furthermore, levels of TNF-α, IL-8 and CRP exhibited significant positive correlation with PMNs%. In conclusion, SCE delayed postpartum ovarian cyclicity in buffaloes. Moreover, TNF-α, IL-8 and CRP assessments could be efficient tools in prediction of SCE in buffaloes.

Hormonal and metabolic profiles related to placental retention with emphasis on oxidative stress and serotonin receptors in pluriparous buffaloes.

This study was designed to investigate the hormonal and metabolic factors associated with placental retention in buffaloes with respect to the roles of oxidative stress biomarkers and serotonin receptors. Blood samples were collected at weeks 3, 2 and 1 pre-partum and at calving from 37 buffaloes; thirty normally dropped their placentae (Non-RFM group) and 7 dropped their placentae after 12 hr post-calving (RFM group). Serum progesterone (P4 ), oestradiol, cortisol, non-esterified fatty acids (NEFA), beta-hydroxybutyric acid (BHBA), antioxidant/oxidant biomarkers and mineral concentrations were assessed. Histopathology and histochemistry were implemented to detect collagen in foetal placental tissues. Immunohistochemistry for serotonin receptors in placental tissues was performed. Significant elevations in P4 , cortisol, NEFA, BHBA and oxidative biomarkers concentrations were observed in the RFM group. However, oestradiol, antioxidants and mineral concentrations were significantly lower in RFM buffaloes than Non-RFM group. Histopathological examination revealed degenerative changes and necrosis in retained placental tissue compared with that in normal placental tissues. Serotonin receptors were significantly expressed with collagen condensation in retained placental tissues. Furthermore, inferior reproductive performance was pronounced in the retained group. In conclusion, retained foetal membranes in buffaloes were associated with hormonal imbalance, metabolic perturbation, oxidative stress, serotonin receptor upregulation and markedly reduced fertility indices.

Associations between metabolic profiles, post-partum delayed resumption of ovarian function and reproductive performance in Egyptian buffalo: Roles of IGF-1 and antioxidants.

This study was conducted on 47 pluriparous pregnant Egyptian buffalo. Body condition score (BCS) was classified and blood samples were collected pre-partum and post-partum for estimation of IGF-1, hormonal, metabolic and antioxidants values. There was palpation per rectum and ultrasonography in addition to quantitation of progesterone (P4) and estradiol-17ß (E-17ß) for monitoring post-partum ovarian resumption. Reproductive indices were calculated 60, 90, 120 and 150 days post-partum. Based on the concentrations of P4 and E-17ß, buffalo were divided into ovulatory and non-ovulatory groups. The P4 and E-17ß were greater (P < 0.001) in ovulatory compared to non-ovulatory buffalo. The BCS and IGF-1 post-partum were greater (P =  0.024; 0.001, respectively) in ovulatory than non-ovulatory buffalo. Glucose and albumin were greater during pre- (P < 0.001; 0.013) and post-partum (P = 0.005; 0.003) periods in ovulatory than non-ovulatory buffalo. Post-partum, NEFA and BHBA concentrations were greater (P < 0.001) in non-ovulatory than ovulatory buffalo. The BUN concentrations were greater (P =  0.002) in non-ovulatory buffalo during pre- and post-partum periods. There were differences in GSH and SOD concentrations between groups (P < 0.001; 0.002, respectively). The BCS, albumin, IGF-1, GSH and SOD concentrations post-partum were negatively correlated with the delay of post-partum ovulation. The post-partum NEFA and BHBA concentrations, however, were positively correlated with delayed post-partum ovulation. Ovulatory buffalo had fewer (P < 0.01) days non-pregnant and for calving intervals as well as greater pregnancy rates than non-ovulatory buffalo. In conclusion, buffalo with delayed post-partum ovarian resumption were prone to have negative energy balance.

Multivitamins preventive therapy against subclinical endometritis in buffaloes: Its correlation to NEFA and oxidative stress.

The current study was designed to elucidate the in vivo antioxidant, preventive, and ameliorating effects of vitamins AD3E on the incidence of subclinical endometritis (SCE) in buffaloes. Twenty-four buffaloes were divided equally into two groups; group I: control and group II: received AD3E combination. Endometrial cytological samples (n=48) were collected using cytobrush to diagnose SCE by counting polymorphonuclear cells (PMN) ≥6% at 5th (W5) and≥4% at 7th (W7) weeks postpartum. Results revealed that serum superoxide dismutase and nitric oxide significantly increased and decreased, respectively at W7 in AD3E group. The PMN% were significantly correlated with oxidative/anti-oxidative stress markers at W5 and W7. Vaginal score, PMN%, and blood neutrophils were significantly higher in the control group buffaloes than the AD3E enriched ones. Therefore, the prevalence of SCE reduced significantly in the AD3E supplemented buffaloes as compared to the control ones at W5 (23.15% and 38.46%) and W7 (9.8% and 32.34%), respectively. The control group revealed higher NEFA levels (P≤0.05) at W5 and W7 than the AD3E group. The AD3E supplemented buffaloes had shorter days open and higher pregnancy rate at 120th and 150th days postpartum than the control ones. In conclusion, micronutrients (AD3E) intervention acts as a safeguard against the incidence of postpartum SCE and significantly improves the reproductive performance of buffaloes.

Ameliorating effects of green tea extract on cadmium induced reproductive injury in male Wistar rats with respect to androgen receptors and caspase- 3.

Cadmium is one of the potent environmental endocrine disruptors that has adverse effects on male fertility. This study was conducted to investigate the protective role of green tea extract (GTE) on cadmium chloride (CdCl2) induced reproductive toxicities in male Wistar rats. Thirty-six male Wistar rats were divided to 4 groups, each one contained 9 rats. Control group, they were gavaged distilled water, meanwhile the treated groups gavaged CdCl2 (3mg/kg), GTE (70mg/kg) and CdCl2+GTE (3mg/kg and 70mg/kg) for 63days. In GTE+ CdCl2 treated rats, the final body weight, relative testicular weight, epididymal weight, seminal and prostate glands weights were significantly (P<0.05) increased compared to CdCl2 intoxicated rats. Sperm cell concentration was significantly (P<0.05) increased in CdCl2+GTE group, while sperm morphological abnormalities were significantly (P<0.05) decreased than CdCl2 group. The levels of superoxide dismutase (SOD) and reduced glutathione (GSH) were elevated significantly (P<0.05) in CdCl2+GTE group, meanwhile catalase activity was significantly (P<0.05) declined than CdCl2 group. Hypercholesterolemia was observed in CdCl2 group than control one while GTE+CdCl2 group revealed significant (P<0.05) reduction in cholesterol level than CdCl2 treated group. Testicular androgen receptors and caspase-3 protein expression showed marked reduction and increase (P<0.05) in CdCl2 group than control, respectively. Treatment with GTE with CdCl2 significantly (P<0.05) increased androgen receptors, while decreased caspase-3 protein than CdCl2 group. In conclusion, GTE exhibited protective effect on testes by inhibiting CdCl2 induced oxidative damage and cellular apoptosis.