The aim of this study was to evaluate the effect of an acute systemic inflammatory response induced by lipopolysaccharide (LPS) in the serum and follicular fluid (FF) high-density lipoprotein (HDL) components, hormone concentrations and granulosa cell gene expression. For this purpose, twenty non-lactating Jersey dairy cows were submitted to a progesterone (P4) - estradiol (E2) based synchronization protocol. Cows received a single i.v. dose of LPS (2.5 μg/kg of body weight) or saline solution (CTL Group) 2 h after P4 insert removal. Blood, granulosa cells and FF samples were collected six hours after LPS injection. Five hours after LPS injection rectal temperature was increased in LPS (P < 0.0001, 40.4 ± 0.1 °C) compared to the CTL cows (38.8 ± 0.1 °C). Serum PON1 activity was reduced by LPS injection (130.2 ± 5.1 vs. 99.6 ± 3.3 U/mL; P < 0.001), as well as HDL-cholesterol concentrations (70.3 ± 5.3 vs. 50.1 ± 6.2 mg/dL; P < 0.05). The FF E2 and P4 concentrations were not different between groups (P > 0.05). The PON1 activity in the FF was also decreased by LPS injection (P = 0.01). In comparison to CTL group, cows injected with LPS had a ten fold reduction in STAR, TLR4 and TNF mRNA expression (P < 0.05). In conclusion, an intravenous LPS challenge in cows induced an acute systemic inflammatory response reducing HDL and its components in serum but not in the FF. Only PON1 activity serum reduction was reflected in the FF in the short term. Additionally, steroidogenic and inflammatory genes had reduced expression in the granulosa cells.
This work examines the effectiveness of a TCG (Tris, citric acid, glucose, 6% egg yolk and 5% glycerol) and a TEST (TES, Tris, glucose, 6% egg yolk and 5% glycerol) sperm extender in the freezing of mouflon spermatozoa at slow cooling rates, using different pre-freezing equilibration times (2-3 hr). It also examines the tolerance of mouflon spermatozoa to different concentrations of cryoprotectants (5, 10, 20% glycerol; 5%, 10%, 20% dimethyl sulfoxide; 6% polyvinylpyrrolidone) and/or sucrose (100, 300, 500 mm). The highest quality (p < .01) thawed spermatozoa were obtained when using the TEST extender and an equilibration time of 3 hr. Sperm motility and membrane integrity were strongly reduced when using rapid freezing rates (60-85°C min ), independent of the concentration of cryoprotectants. The lowest sucrose concentration (100 mm) provided the highest (p < .05) percentage of motile spermatozoa and live spermatozoa with an intact acrosome. Vitrified-warmed sperm variables were at their best when the spermatozoa was diluted in TCG-6% egg yolk + 100 mm sucrose and warmed at 60°C. Slow warming at 37°C strongly reduced (p < .05) sperm motility and viability. However, sperm vitrification returned lower fertility, sperm motility and sperm viability values than conventional sperm freezing.
Caloric restriction (CR) increases the preservation of the ovarian primordial follicular reserve, which can potentially delay menopause. Rapamycin also increases preservation on the ovarian reserve, with similar mechanism to CR. Therefore, the aim of our study was to evaluate the effects of rapamycin and CR on metabolism, ovarian reserve, and gene expression in mice. Thirty-six female mice were allocated into three groups: control, rapamycin-treated (4 mg/kg body weight every other day), and 30% CR. Caloric restricted females had lower body weight (P < 0.05) and increased insulin sensitivity (P = 0.003), while rapamycin injection did not change body weight (P > 0.05) and induced insulin resistance (P < 0.05). Both CR and rapamycin females displayed a higher number of primordial follicles (P = 0.02 and 0.04, respectively), fewer primary, secondary, and tertiary follicles (P < 0.05) and displayed increased ovarian Foxo3a gene expression (P < 0.05). Despite the divergent metabolic effects of the CR and rapamycin treatments, females from both groups displayed a similar increase in ovarian reserve, which was associated with higher expression of ovarian Foxo3a.
This work examines the effects of subsequent cycles of freezing-thawing on giant panda (Ailuropoda melanoleuca) sperm morphometry and function, and assesses whether density-gradient centrifugation (DGC) can increase the number of freezing-thawing cycles this sperm can withstand. A sperm sample was collected by electroejaculation from a mature giant panda and subjected to five freezing-thawing cycles. Although repeated freezing-thawing negatively affected (P < 0.05) sperm motility and membrane integrity, in both nonselected and DCG-selected sperm samples, >60% of the sperm cells in both treatments showed acrosome integrity even after the fifth freezing cycle. In fresh semen, the sperm head length was 4.7 μm, the head width 3.6 μm, area 14.3 μm(2) and perimeter length 14.1 μm. The present results suggest that giant panda sperm trends to be resistant to repeated freezing-thawing, even without DGC selection.
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