Maternal reproductive performance and fetal development of the Wistar Audiogenic Rat (WAR) strain.

Wistar Audiogenic Rat (WAR) strain is an animal model for epilepsy studies, the chronic multifactorial disease that affects millions of people worldwide. The animals of this strain are genetically predisposed to sound-induced seizures, called audiogenic seizures, and have been used for many years in studies to understand the mechanisms involved in the epilepsies and their neuropsychiatric comorbidities, as well as the screening of potential anti-convulsant agents. Nevertheless, little is known about the reproductive characteristics of these animals. The main goal of this study was to characterize the female reproductive performance and the fetal growth of WARs in comparison to the Wistar rats, obtaining important information for physiology and behavioral studies, as well as for the preservation of the strain. The results indicated few differences between WAR and Wistar regarding the female reproductive performance. There was no significant difference in the number of pregnant females by mating, number of live births per female, number of cells per blastocyst, and several characteristics related to reproductive performance, such as pre- and post-implantation losses. However, significant differences were observed in birth weight and weight gain until weaning, with WAR animals presenting a body weight below Wistar at birth and reduced body weight gain during the lactation period. In addition, the WAR females showed lower body weight on the day 20 of pregnancy and a larger number of corpora lutea, when compared with those of Wistar animals. Thus, we conclude that although Wistar and WAR strains have few differences in their reproductive performance, which might impact future physiological life challenges or others experimentally induced procedures, it still is a very viable strain regarding reproduction. Abbreviations: CONCEA: National Council for the Control of Animal Experimentation; GEPR: genetically epilepsy-prone rats; WAR: Wistar Audiogenic Rat.

Elevated circulatingadenosine deaminase activity in women with preeclampsia: association with pro-inflammatory cytokine production and uric acid levels.

OBJECTIVE: Preeclampsia is a specific disorder of human pregnancy that is associated with hyperuricemia and higher levels of pro-inflammatory cytokines. Adenosine deaminase (ADA) is an enzyme present in all human tissues, and is considered an indicator of cellular inflammation. In the present study we assess whether adenosine deaminase (ADA) activity is altered in women with preeclampsia (PE) and contributes to elevated levels of uric acid and pro-inflammatory cytokine production. STUDY DESIGN: The population studied consisted of 60 women with PE, 30 normotensive pregnant women (NT) and 20 non-pregnant women (NP). Uric acid concentration and ADA activity were determined in the serum. Peripheral blood mononuclear cells (PBMCs) were isolated and evaluated for intracellular nuclear transcription factor kappa B (NF-κB) levels and for endogenous tumor necrosis factor-α (TNF-α) and interleukin-1ß (IL-1ß) production. The data were evaluated with parametric or non-parametric tests with significance set at P<0.05. RESULTS: ADA levels were higher in the PE group compared with the NT and NP groups (P<0.001). A positive correlation between ADA and uric acid levels was identified in women with PE (P<0.001). Endogenous production of IL-1ß and TNF-α, as well as intracellular NF-κB levels, were higher in PBMCs from the PE group than from NT and NP women (P<0.01) and correlated with the ADA concentration in preeclamptic women (P<0.01). CONCLUSION: An elevation in ADA activity in women with PE may contribute to their increased levels of uric acid and pro-inflammatory immune activity.

Low versus atmospheric oxygen tension for embryo culture in assisted reproduction: a systematic review and meta-analysis.

OBJECTIVE: To appraise the available evidence comparing low oxygen (LowO2) and atmospheric oxygen tension (AtmO2) for embryo culture. DESIGN: Systematic review and meta-analysis. SETTING: Not applicable. PATIENT(S): Women undergoing assisted reproduction using embryo culture. INTERVENTION(S): Embryo culture using LowO2 versus AtmO2. MAIN OUTCOME MEASURE(S): Reproductive, laboratory, and pregnancy outcomes. RESULT(S): A total of 21 studies were included in this review. All used O2 concentration between 5% and 6% in the LowO2 group. Considering the studies that randomized women/couples, we observed very low quality evidence that LowO2 is better for live birth/ongoing pregnancy (relative risk [RR] = 1.1, 95% confidence interval [CI] 1.0-1.3) and clinical pregnancy (RR = 1.1, 95% CI 1.0-1.2). Considering the studies that randomized oocytes/embryos, we observed low quality evidence of no difference of fertilization (RR = 1.0, 95% CI 1.0-1.0) and cleavage rate (RR = 1.0, 95% CI 1.0-1.1), and low quality evidence that LowO2 is better for high/top morphology at the cleavage stage (RR = 1.2, 95% CI 1.1-1.3). No studies comparing pregnancy outcomes were identified. Several studies used different incubators in the groups-a new model for the LowO2 group and an old model for the AtmO2 group. The risk of detection bias for the laboratory outcomes was high as embryologists were not blinded. CONCLUSION(S): Although we observed a small improvement (∼5%) in live birth/ongoing pregnancy and clinical pregnancy rates (PRs), the evidence is of very low quality and the best interpretation is that we are still very uncertain about differences in this comparison. The clinical equipoise remains and more large well-conducted randomized controlled trials are needed. They should use the same incubators in both groups and the embryologists should be blinded at least when evaluating laboratory outcomes.

N-Acetyl-Cysteine and l-Carnitine Prevent Meiotic Oocyte Damage Induced by Follicular Fluid From Infertile Women With Mild Endometriosis.

This study evaluated the potential protective effect of the antioxidants, l-carnitine (LC) and N-acetyl-cysteine (NAC), in preventing meiotic oocyte damage induced by follicular fluid (FF) from infertile women with mild endometriosis (ME). We performed an experimental study. The FF samples were obtained from 22 infertile women undergoing stimulated cycles for intracytoplasmic sperm injection (11 with ME and 11 without endometriosis). Immature bovine oocytes were submitted to in vitro maturation (IVM) divided into 9 groups: no-FF (No-FF); with FF from control (CFF) or ME (EFF) groups; and with LC (C + LC and E + LC), NAC (C + NAC and E + NAC), or both antioxidants (C + 2Ao and E + 2Ao). After IVM, oocytes were immunostained for visualization of microtubules and chromatin by confocal microscopy. The percentage of meiotically normal metaphase II (MII) oocytes was significantly lower in the EFF group (51.35%) compared to No-FF (86.36%) and CFF (83.52%) groups. The E + NAC (62.22%), E + LC (80.61%), and E + 2Ao (61.40%) groups showed higher percentage of normal MII than EFF group. The E + LC group showed higher percentage of normal MII than E + NAC and E + 2Ao groups and a similar percentage to No-FF and CFF groups. Therefore, FF from infertile women with ME causes meiotic abnormalities in bovine oocytes, and, for the first time, we demonstrated that the use of NAC and LC prevents these damages. Our findings elucidate part of the pathogenic mechanisms involved in infertility associated with ME and open perspectives for further studies investigating whether the use of LC could improve the natural fertility and/or the results of in vitro fertilization of women with ME.

Silibinin modulates the NF-κb pathway and pro-inflammatory cytokine production by mononuclear cells from preeclamptic women.

Preeclampsia (PE) is a complication of human pregnancy associated with an intense inflammatory response involving leukocyte activation, as well as elevated production of pro-inflammatory cytokines. The nuclear transcription factor-kappa B (NF-κB) is present in cells of the immune system and is responsible for transcription of genes coding for pro-inflammatory proteins. Silibinin is the main component of silymarin, a polyphenolic extract obtained from fruits and seeds of Silybum marianum with potent hepatoprotective and anti-inflammatory activities. In this study, we assessed whether silibinin modulated NF-κB activity and the production of pro-inflammatory cytokines by peripheral blood mononuclear cells (PBMCs) from preeclamptic patients. PBMC from women with PE, normotensive (NT) pregnant women, and nonpregnant (NP) women were cultured with or without silibinin (5 µM and 50 µM) and 1 µg/mL lipopolysaccharide (LPS) for 18 h. The supernatants were assayed for tumor necrosis factor-alpha (TNF-α) and interleukin-1 beta (IL-1ß) by ELISA. Cells were cultured for 30 min to evaluate NF-κB activity. There was increased endogenous activation of NF-κB as well as TNF-α and IL-1ß release by PBMC in the PE group compared with the NT and NP groups. A positive correlation between NF-κB activity and cytokine production was also observed in the PE group. Silibinin was capable of reducing, at least in part, the levels of NF-κB and cytokines TNF-α and IL-1ß in preeclamptic women. We conclude that silibinin exhibits potent anti-inflammatory activity on PBMC from preeclamptic women by downmodulation of NF-κB activation and inflammatory cytokine production.