Maternal exposure to an enrichment environment promotes uterine vascular remodeling and prevents embryo loss in mice.

Aim: Implantation-related events are crucial for pregnancy success. In particular, defects in vascular remodeling at the maternal-fetal interface are associated with spontaneous miscarriage and recurrent pregnancy loss. Physical activity and therapies oriented to reduce stress improve pregnancy outcomes. In animal models, environmental stimulation and enrichment are associated with enhanced well-being, cognitive function and stress resilience. Here we studied whether exposure of BALB/c mice to an enriched environment (EE) regulates crucial events during early gestation at the maternal-fetal interface. Method: Pregnant BALB/c mice were exposed to the EE that combines non-invasive stimuli from the sensory pathway with voluntary physical activity. The pregnancy rate was evaluated. Implantation sites were investigated microscopically and macroscopically. Vascular adaptation parameters at the maternal-fetal interface were analyzed. Results: We found that exposure to the EE prevented pregnancy loss between gestational days 7 and 15. Also, it increased the diameter of the uterine artery and decreased the wall:lumen ratio of the mesometrial decidual vessels, suggesting that EE exposure promotes vascular remodeling. Moreover, it increased nitric oxide synthase activity and inducible nitric oxide synthase expression, as well as prostaglandin F2α production and endoglin expression in the implantation sites. Conclusion: Exposure of pregnant females to the EE regulates uterine physiology, promoting vascular remodeling during early gestation. These adaptations might contribute to preventing embryo loss. Our results highlight the importance of the maternal environment for pregnancy success. The design of an “EE-like” protocol for humans could be considered as a new non-pharmacologic strategy to prevent implantation failure and recurrent miscarriage.

Disruption of Folate Metabolism Causes Poor Alignment and Spacing of Mouse Conceptuses for Multiple Generations

Abnormal uptake or metabolism of folate increases risk of human pregnancy complications, though the mechanism is unclear. Here, we explore how defective folate metabolism influences early development by analysing mice with the hypomorphic Mtrrgt mutation. MTRR is necessary for methyl group utilisation from folate metabolism, and the Mtrrgt allele disrupts this process. We show that the spectrum of phenotypes previously observed in Mtrrgt/gt conceptuses at embryonic day (E) 10.5 is apparent from E8.5 including developmental delay, congenital malformations, and placental phenotypes. Notably, we report misalignment of some Mtrrgt conceptuses within their implantation sites from E6.5. The degree of misorientation occurs across a continuum, with the most severe form visible upon gross dissection. Additionally, some Mtrrgt/gt conceptuses display twinning. Therefore, we implicate folate metabolism in blastocyst orientation and spacing at implantation. Skewed growth likely influences embryo development since developmental delay and heart malformations (but not defects in neural tube closure or trophoblast differentiation) associate with severe misalignment of Mtrrgt/gt conceptuses. Typically, the uterus is thought to guide conceptus orientation. To investigate a uterine effect of the Mtrrgt allele, we manipulate the maternal Mtrr genotype. Misaligned conceptuses were observed in litters of Mtrr+/+, Mtrr+/gt, and Mtrrgt/gt mothers. While progesterone and/or BMP2 signalling might be disrupted, normal decidual morphology, patterning, and blood perfusion are evident at E6.5 regardless of conceptus orientation. These observations argue against a post-implantation uterine defect as a cause of conceptus misalignment. Since litters of Mtrr+/+ mothers display conceptus misalignment, a grandparental effect is explored. Multigenerational phenotype inheritance is characteristic of the Mtrrgt model, though the mechanism remains unclear. Genetic pedigree analysis reveals that severe conceptus skewing associates with the Mtrr genotype of either maternal grandparent. Moreover, the presence of conceptus skewing after embryo transfer into a control uterus indicates that misalignment is independent of the peri- and/or post-implantation uterus and instead is likely attributed to an embryonic mechanism that is epigenetically inherited. Overall, our data indicates that abnormal folate metabolism influences conceptus orientation over multiple generations with implications for subsequent development. This study casts light on the complex role of folate metabolism during development beyond a direct maternal effect.

Ectopic Pregnancy in Tigray, Ethiopia: A Cross-Sectional Survey of Prevalence, Management Outcomes, and Associated Factors

Background. Ectopic pregnancy is a neglected and challenging gynecologic problem in developing countries including Ethiopia. Objective. The present study is aimed at assessing the prevalence of ectopic pregnancy, its management outcomes, and factors associated with management outcomes in Tigray, North Ethiopia. Methods. We employed a four-year retrospective cross-sectional study from September 2015 to August 2019. We extracted data about all pregnant mothers who were admitted and managed for EPs in Axum, Tigray. Ectopic pregnancy and its outcomes (favorable and unfavorable) were the dependent variables, and age, residence, ethnicity, religion, parity, history of abortion, history of EP, pelvic infections, history of surgical procedures, and use contraceptives were the independent variables. We employed descriptive statistics and bivariate and multivariate logistic regression analyses using SPSS. Ethical clearance was obtained from Axum University, Tigray, Ethiopia. Results. The overall prevalence of ectopic pregnancy was 0.52% of total deliveries, which equates to 1 : 193 deliveries. Surgery for ectopic pregnancy accounts for 7.6% of all gynecological surgeries. Most participants were in the age group 26–30 years and lived in rural areas. Among the different EP implantation sites, most cases (92.4%) occurred in the fallopian tube, followed by 5.1% in the ovary and 2.5% in abdominal EPs. Surgical management (laparotomy) was undertaken for all the 79 women diagnosed with EPs, including laparotomy (100%), salpingo-oophorectomy (17.7%), salpingectomy (73.9%), oophorectomy (3.4%), cornual resection (2.5%), and removal of concepts tissue 2.5. The record reports that intraoperative procedure was correctly managed for 47 (59.5%) women but the condition of EP procedure was ruptured for about two-thirds (63.3%) of the women. Thirty (38%) patients had developed some complications after surgery including anemia ( hemoglobin < 10.5 ) ( n = 12 ), fever ( n = 10 ), wound infection ( n = 2 ), and pneumonia ( n = 2 ). Women who were from urban ( AOR = 11.2 , 95% CI: 2.65-47.2) and who had normal hemoglobin at presentation ( AOR = 9.94 , 95% CI: 2.03-48.7) were associated with favorable maternal outcomes. Conclusions. More than one-third of women with ectopic pregnancies had an unfavorable maternal outcome, which was higher among rural residents and anemic mothers. Women living in rural areas and anemia during pregnancy should seek special attention in the management of EPs. We also recommend improving the data management of hospitals in Ethiopia.

New Stimulation Device to Drive Multiple Transverse Intrafascicular Electrodes and Achieve Highly Selective and Rich Neural Responses

Peripheral Nerve Stimulation (PNS) is a promising approach in functional restoration following neural impairments. Although it proves to be advantageous in the number of implantation sites provided compared with intramuscular or epimysial stimulation and the fact that it does not require daily placement, as is the case with surface electrodes, the further advancement of PNS paradigms is hampered by the limitation of spatial selectivity due to the current spread and variations of nerve physiology. New electrode designs such as the Transverse Intrafascicular Multichannel Electrode (TIME) were proposed to resolve this issue, but their use was limited by a lack of innovative multichannel stimulation devices. In this study, we introduce a new portable multichannel stimulator—called STIMEP—and implement different stimulation protocols in rats to test its versatility and unveil the potential of its combined use with TIME electrodes in rehabilitation protocols. We developed and tested various stimulation paradigms in a single fascicle and thereafter implanted two TIMEs. We also tested its stimulation using two different waveforms. The results highlighted the versatility of this new stimulation device and advocated for the parameterizing of a hyperpolarizing phase before depolarization as well as the use of small pulse widths when stimulating with multiple electrodes.

Effects of Aurora kinase A on mouse decidualization via Stat3-plk1-cdk1 pathway

Background Decidualization is essential to the successful pregnancy in mice. The molecular mechanisms and effects of Aurora kinase A (Aurora A) remain poorly understood during pregnancy. This study is the first to investigate the expression and role of Aurora A during mouse decidualization. Methods Quantitative real time polymerase chain reaction, western blotting and in situ hybridization were used to determine the expression of Aurora A in mouse uteri. Aurora A activity was inhibited by Aurora A inhibitor to explore the role of Aurora A on decidualization via regulating the Aurora A/Stat3/Plk1/Cdk1 signaling pathway. Results Aurora A was strongly expressed at implantation sites compared with inter-implantation sites. Furthermore, Aurora A was also significantly increased in oil-induced deciduoma compared with control. Both Aurora A mRNA and protein were significantly increased under in vitro decidualization. Under in vitro decidualization, Prl8a2, a marker of mouse decidualization, was significantly decreased by TC-S 7010, an Aurora A inhibitor. Additionally, Prl8a2 was reduced by Stat3 inhibitor, Plk1 inhibitor and Cdk1 inhibitor, respectively. Moreover, the protein levels of p-Stat3, p-Plk1 and p-Cdk1 were suppressed by TC-S 7010. The protein levels of p-Stat3, p-Plk1 and p-Cdk1 were also suppressed by S3I-201, a Stat3 inhibitor). SBE 13 HCl (Plk1 inhibitor) could reduce the protein levels of p-Plk1 and p-Cdk1. Collectively, Aurora A could regulate Stat3/Plk1/Cdk1 signaling pathway. Conclusion Our study shows that Aurora A is expressed in decidual cells and should be important for mouse decidualization. Aurora A/Stat3/Plk1/Cdk1 signaling pathway may be involved in mouse decidualization.

Evaluation of the Reproductive Ability of Male Rats in Acute Toxoplasmosis

Toxoplasmosis is a parasitic disease of humans and animals caused by Toxoplasma gondii. Toxoplasma is an intracellular parasite that belongs to the simplest and has a complex development cycle. Infection with Toxoplasma is possible orally, transplacentally, percutaneously (if the integrity of the skin is damaged). This invasion is often the cause of problems with bearing pregnancy, as well as the development of congenital anomalies in children. The purpose of the study was to study the reproductive ability of male rats in acute toxoplasmosis. Materials and methods. The experiment was performed on 90 female and 45 male Wistar rats with a body weight of 180-200 g. The intact control males were orally injected with 2 ml of 0.2% starch gel. Experimental groups of males were infected with an invasive Toxoplasma gondii culture at a dose of 25 tachyzoites per 1 g of body weight (5000 tachyzoites per rat) and 50 tachyzoites per 1 g of body weight (10000 tachyzoites per rat). Then the males of all groups were coupled with the females for 3 days. The effect of toxoplasmas on the reproductive ability of male rats was assessed by the development of pregnancy and changes in the levels of pre- and post-implantation embryo death in female rats on the 7th, 14th, and 21st days after pregnancy. To account for changes in the pre- and post-implantation death of embryos in female rats after removal from the experiment, the uterus and ovaries were isolated, the uterine horns were opened, the number of implantation sites, the total number of embryos, the number of living and dead embryos, the number of resorption, and the number of yellow bodies in the ovaries were determined. Results and discussion. In the females of the 4th, 5th and 6th groups (coupling with males infected at the dose of 25 tachyzoites per 1 g of body weight), a decrease in the number of implantation sites in the uterus, the total number of embryos and the number of live embryos was recorded by 1.8-1.9 times compared to the control parameters. In female rats of the 7th, 8th and 9th groups (coupling with males infected at the dose of 50 tachyzoites per 1 g of body weight), there was a decrease in the number of implantation sites in the uterus, the total number of embryos and the number of live embryos by 5.6-6.8 times compared to the control. When compared to the results obtained from the females of the 4th, 5th and 6th groups, a decrease in these indicators was recorded by 3.1-3.5 times. Conclusion. Toxoplasma gondii has an effect on reproductive capacity in male rats expressed in changes of the levels of preimplantation mortality in female rats. The obtained effect depends on the dose of infection and the period of parasitosis development in males

Abstract P224: Obesity Is Associated With Hypoxia At The Maternal-fetal Interface In A Mouse Model Of Superimposed Preeclampsia, BPH/5

Obesity impacts 1/3 of US adults. Maternal obesity significantly increases risk of adverse pregnancy outcomes, including preeclampsia (PE). Late-gestational hypertension and fetal growth restriction (FGR), hallmarks of PE, are observed spontaneously in BPH/5 mice. Similar to obese preeclamptic women, BPH/5 have increased visceral white adipose tissue (WAT) and are leptin resistant. We hypothesize that attenuation of maternal obesity and reduction of reproductive WAT in pregnant BPH/5 female mice will improve decidualization, placental development, and attenuate FGR. To test this hypothesis, pregnant C57 and BPH/5 female mice fed ad libitum (lib) and pair-fed (PF) BPH/5 female mice, beginning at day of copulatory plug detection, e0.5 (n=5/group), were utilized. Pair-feeding BPH/5 females the equivalent daily food intake of lean control ad lib fed C57 females reduced maternal WAT and circulating leptin in this model by e7.5 as well as reduced pro-inflammatory cytokines, Ptgs2 and IL-6, mRNA in the decidua. Therefore, we tested whether hypoxia-related genes, hypoxia inducible factor (Hif) 1α, heme oxygenase-1 (Ho-1), and stem cell factor (Scf), would be altered in the decidua of PF BPH/5 at e7.5. Using real time PCR, we found that e7.5 implantation sites from ad lib fed BPH/5 had a 1.5 to 2-fold increase in Hif1α, Ho-1, Scf, and VEGF mRNA (p<0.05) compared to C57 that was significantly reduced by PF. Furthermore, real time PCR for leptin (lep) mRNA showed a 6-fold increase in e7.5 implantation sites from ad lib fed BPH/5 versus C57 and lep along with lep receptor mRNA was reduced in PF BPH/5 (p<0.05). Therefore, PF BPH/5 pregnant mice have attenuated obesity and leptin in WAT, circulation, and e7.5 implantation sites that is associated with a reduction in markers of hypoxia at the maternal-fetal interface. Finally, BPH/5 PF litter sizes are significantly higher than ad lib fed, 6 vs 3.5, respectively (n=5-8; p<0.05) and symmetrical FGR is attenuated. In conclusion, maternal weight loss in BPH/5 beginning at conception may improve placental development by mitigating hypoxia at the maternal-fetal interface in this model. Future investigations are needed to determine this effect on the maternal hypertensive syndrome and offspring outcomes long-term.

Osteointegration of polylactide-based implants

Background. Materials degrading after implantation into bone are in the field of actual vision of orthopediс surgeon. These materials include polylactides, which are the ideal material for creating bone implants in 3D-printer, especially implants of complex shapes and different sizes. The purpose of the study is to conduct a comparative analysis of bone remodeling under conditions of implantation of polylactide 3D-printed screws into metadiaphyseal and diaphyseal defects of the rat femur. Materials and methods. Comparative analysis of bone remodeling under conditions of implantation of polylactide Ingeo™ Biopolymer 4032D 3D-printed screws into the metadiaphyseal and diaphyseal defects of the rat femur for 15, 30, 90, 180 and 270 days are conducted. After implantation of polylactide, areas of bone with implants were examined by the histological method with the determination of the osteointegration index. Results. It was found that for all the study periods the implants kept their shape, were surrounded by bone tissue. The osseointegration index on the 270th day in metadiaphyseal and diaphyseal defects is 97.1 and 94.3 %, respectively, and is statistically higher compared to the 15th day by 2.2 and 2.3 times (p < 0.001). Conclusions. The polylactide-based Ingeo™ Biopolymer 4032D implants are biocompatible, have high osteointegration qualities, do not cause inflammation in the surrounding soft tissues and bone marrow, do not lead to destructive changes of the bone in the implantation sites. At the end of the study (270 days), the degradation of polylactide is not found, which makes it possible to use it for fixation or filling cavities in compact and spongy bones for a long time.

The multistep process of vaginal cancer arising from deep infiltrating endometriosis: a case report

Background Malignant transformation of endometriosis in extraovarian sites remains rare. Furthermore, the process is not definitely understood. Case presentation Herein, we report the case of a 40-year-old premenopausal nulligravida woman who presented with vaginal bleeding and who was finally diagnosed with a vaginal cancer originating from endometriosis and with a synchronous endometrial cancer. A gynecologic examination revealed a multiple polypoid mass on the posterior vaginal fornix. Magnetic Resonance Imaging of the pelvis showed two masses abutting respectively on the anterior uterine wall, and in the rectovaginal septum. The patient underwent a total laparoscopic excision of the rectovaginal mass, radical hysterectomy and low anterior resection of the rectum. The lesions were diagnosed as endometriosis, endometriosis-associated complex hyperplasia and endometrioid cancer. Furthermore, a synchronous endometrioid endometrial cancer was reported. Conclusions This case revealed the multistep process of malignant transformation of deep infiltrating endometriosis. The progression was individualized between implantation sites and in the same organ.