Evaluation of progestogen supplementation for luteal phase support in fresh in vitro fertilization cycles

© 2019 American Society for Reproductive Medicine Objective: To evaluate the effectiveness of progestogen supplementation in improving clinical pregnancy rates in women undergoing fresh IVF cycles and to compare different routes, start times, durations, and estrogen coadministration regimen. Design: Comprehensive systematic review and meta-analysis. Setting: University. Patient(s): Women undergoing fresh IVF cycles who did and did not receive progestogen supplementation. Intervention(s): Summary odds ratios (ORs) were calculated by binomial logistic regression. Main Outcome Measure(s): Clinical pregnancy rates. Result(s): Eighty-two articles (26,726 women) were included. Clinical pregnancy rates were increased by IM (OR = 4.57), vaginal (OR = 3.34), SC (OR = 3.36), or oral (OR = 2.57) progestogen supplementation versus no treatment. The greatest benefit was observed when progestogens were supplemented IM versus vaginally (OR = 1.37). The optimal time to commence administration was between oocyte retrieval and ET (OR = 1.31), with oocyte retrieval +1 day being most beneficial. Coadministration of estrogen had no benefit (OR = 1.33), whether progestogens were coadministered vaginally or IM. Clinical pregnancy rates were equivalent when progestogen supplementation was ceased after ≤3 weeks or continued for up to 12 weeks (OR = 1.06). Conclusion(s): This broad-ranging meta-analysis highlights the need to reevaluate current clinical practice. The use of progestogens in fresh IVF cycles is substantially beneficial to clinical pregnancy. Critically, the use of IM progestogens should not be dismissed, as it yielded the greatest clinical pregnancy rates. Pregnancy success was impacted by initiation of therapy, with 1 day after oocyte retrieval being optimal. There is little evidence to support coadministration of estrogen or prolonging progestogen treatment beyond 3 weeks

Gene expression profiling of breast tumours from New Zealand patients

AIMS: New Zealand has one of the highest rates of breast cancer incidence in the world. We investigated the gene expression profiles of breast tumours from New Zealand patients, compared them to gene expression profiles of international breast cancer cohorts and identified any associations between altered gene expression and the clinicopathological features of the tumours. METHODS: Affymetrix microarrays were used to measure the gene expression profiles of 106 breast tumours from New Zealand patients. Gene expression data from six international breast cancer cohorts were collated, and all the gene expression data were analysed using standard bioinformatic and statistical tools. RESULTS: Gene expression profiles associated with tumour ER and ERBB2 status, molecular subtype and selected gene expression signatures within the New Zealand cohort were consistent with those found in international cohorts. Significant differences in clinicopathological features such as tumour grade, tumour size and lymph node status were also observed between the New Zealand and international cohorts. CONCLUSIONS: Gene expression profiles, which are a sensitive indicator of tumour biology, showed no clear di¬fference between breast tumours from New Zealand patients and those from non-New Zealand patients. This suggests that other factors may contribute to the high and increasing breast cancer incidence in New Zealand compared to international populations

Transforming growth factor-β1 disrupts angiogenesis during the follicular–luteal transition through the Smad–serpin family E member 1 (SERPINE1)/serpin family B member 5 (SERPINB5) signalling pathway in the cow

Intense angiogenesis is critical for the development of the corpus luteum and is tightly regulated by numerous factors. However, the exact role transforming growth factor beta 1 (TGFB1) plays during this follicular-luteal transition remains unclear. This study hypothesized that TGFB1 acting through TGFBR1 and Smad2/3 signaling would suppress angiogenesis during the follicular-luteal transition. Using a serum-free luteinizing follicular angiogenesis culture system, TGFB1 (1 and 10ng.mL-1) markedly disrupted the formation of capillary-like structures, reducing endothelial cell network area and number of branch points (P[less than]0.001). Furthermore, TGFB1 activated canonical Smad signaling and inhibited endothelial nitric oxide synthase (NOS3) mRNA expression, but up-regulated latent TGF-beta binding protein, type I TGFB receptor (TGFBR1), SERPINE1 and SERPINB5 mRNA expression. TGFBR1 inhibitor, SB431542, reversed the SERPINE1 and SERPINB5 up-regulation by TGFB1. Additionally, TGFB1 reduced progesterone synthesis through decreasing STAR, CYP11A1 and HSD3B1 expression. These results show that TGFB1 regulated NOS3, SERPINE1, and SERPINB5 expression via TGFBR1 and Smad2/3 signaling and could be the mechanism by which TGFB1 suppresses endothelial networks. Thereby, TGFB1 may provide a critical homeostatic control of angiogenesis during the follicular-luteal transition. Our findings reveal the molecular mechanisms underlying the actions of TGFB1 in early luteinization which may lead to novel therapeutic strategies to reverse luteal inadequacy

Luteal angiogenesis and its control

Angiogenesis, the formation of new blood vessels from pre-existing ones, is critical to luteal structure and function; In addition, it is a complex and tightly regulated process. Not only does rapid and extensive angiogenesis occur to provide the corpus luteum (CL) with an unusually high blood flow and support its high metabolic rate, but in the absence of pregnancy the luteal vasculature must rapidly regress to enable the next cycle of ovarian activity. This review describes a number of the key endogenous stimulatory and inhibitory factors, which act in a delicate balance to regulate luteal angiogenesis and ultimately luteal function. In vitro luteal angiogenesis cultures have demonstrated critical roles for fibroblast growth factor 2 (FGF2) in endothelial cell proliferation and sprouting, whilst other factors such as vascular endothelial growth factor (VEGFA) and platelet derived growth factor (PDGF) were important modulators in the control of luteal angiogenesis. Post-transcriptional regulation by small non-coding micro-RNAs, is also likely to play a central role in the regulation of luteal angiogenesis. Appropriate luteal angiogenesis requires the coordinated activity of numerous factors expressed by several cell types at different times and this review will also describe the role of perivascular pericytes and the importance of vascular maturation and stability. It is hoped that a better understanding of the critical processes underlying the transition from follicle to CL, and subsequent luteal development will benefit the management of luteal function in the future

Maternal protein restriction affects fetal ovary development in sheep

Maternal malnutrition has important developmental consequences for the foetus. Indeed, adverse fetal ovarian development could have lifelong impact, with potentially reduced ovarian reserve and fertility of the offspring. This study investigated the effect of maternal protein restriction on germ cell and blood vessel development in the fetal sheep ovary. Ewes were fed control (n = 7) or low protein (n = 8) diets (17.0 g vs 8.7 g crude protein/MJ metabolizable energy) from conception to day 65 of gestation (gd65). On gd65, fetal ovaries were subjected to histological and immunohistochemical analysis to quantify germ cells (OCT4, VASA, DAZL), proliferation (Ki67), apoptosis (caspase 3) and vascularisation (CD31). Protein restriction reduced the fetal ovary weight (P 0.05). The density of germ cells was unaffected by maternal diet (P > 0.05). In the ovarian cortex, OCT4+ve cells were more abundant than DAZL+ve (P 0.05). Similarly, maternal protein restriction had no effect on germ cell proliferation or apoptotic indices (P > 0.05) and the number, area and perimeter of medullary blood vessels and degree of microvascularisation in the cortex (P > 0.05). In conclusion, maternal protein restriction decreased ovarian weight despite not affecting germ cell developmental progress, proliferation, apoptosis, or ovarian vascularity. This suggests that reduced maternal protein has the potential to regulate ovarian development in the offspring

Detrimental effects of uterine disease and lipopolysaccharide on luteal angiogenesis

© 2020 Society for Endocrinology Reproductive tract inflammatory disease (RTID) commonly occurs after the traumatic events of parturition and adversely affects follicular function. This study is the first to describe the cellular and steroidogenic characteristics of corpora lutea from cattle with RTID and the effects of pathogen-associated molecular patterns (PAMPs) on luteal angiogenesis and function in vitro. Luteal weight (P 3-fold lower in RTID cows (P 0.05). Moreover, the adverse effect of LPS on luteal EC networks was dose-dependent and effective from 1 ng/mL (P < 0.05), while few EC networks were present above 10 ng/mL LPS (P < 0.001). LPS reduced proliferation (P < 0.05) and increased apoptosis of EC (P < 0.001). The specific TLR4 inhibitor TAK242 reversed the effects of LPS on EC networks. In conclusion, luteal vasculature is adversely sensitive to LPS acting via TLR4, therefore ovarian exposure to LPS from any Gram-negative bacterial infection will profoundly influence subsequent reproductive potential

A randomised controlled trial to evaluate the impact of indoor living space on dairy cow production, reproduction and behaviour

As a global society, we have a duty to provide suitable care and conditions for farmed livestock to protect animal welfare and ensure the sustainability of our food supply. The suitability and biological impacts of housing conditions for intensively farmed animals is a complex and emotive subject, yet poorly researched, meaning quantitative evidence to inform policy and legislation is lacking. Most dairy cows globally are housed for some duration during the year, largely when climatic conditions are unfavourable. However, the impact on biology, productivity and welfare of even the most basic housing requirement, the quantity of living space, remains unknown. We conducted a long-term (1-year), randomised controlled trial (CONSORT 10 guidelines) to investigate the impact of increased living space (6.5m2 vs 3m2 per animal) on critical aspects of cow biology, behaviour and productivity. Adult Holstein dairy cows (n = 150) were continuously and randomly allocated to a high or control living space group with all other aspects of housing remaining identical between groups. Compared to cows in the control living space group, cows with increased space produced more milk per 305d lactation (primiparous: 12,235L vs 11,592L, P < 0.01; multiparous: 14,746L vs 14,644L, P < 0.01) but took longer to become pregnant after calving (primiparous: 155d vs 83d, P = 0.025; multiparous: 133d vs 109d). In terms of behaviour, cows with more living space spent significantly more time in lying areas (65min/d difference; high space group: 12.43h/day, 95% CI = 11.70-13.29; control space group: 11.42h/day, 95% CI = 10.73-12.12) and significantly less time in passageways (64min/d), suggesting enhanced welfare when more space was provided. A key physiological difference between groups was that cows with more space spent longer ruminating each day. This is the first long term study in dairy cows to demonstrate that increased living space results in meaningful benefits in terms of productivity and behaviour and suggests that the interplay between farmed animals and their housed environment plays an important role in the concepts of welfare and sustainability of dairy farming

Molecular Regulation of Noradrenaline in Bovine Corpus Luteum

Noradrenergic stimulation increases progesterone, oxytocin and prostaglandins in the bovine luteal tissue. Better understanding of noradrenaline (NA) role in bovine the corpus luteum (CL) can advance our current knowledge on the regulatory mechanism of CL function. The present study was conducted to explore the source of noradrenaline and further to investigate whether nerve growth factor (NGF), insulin like growth factor 1 (IGF1) and transforming growth factor b1 (TGFb1) influence the expression of dopamine-b-hydroxylase (DBH), biosynthetic enzyme of NA in cultured bovine luteal cells. Corpora lutea were collected and classified into stages of early, developing, mid, late, and regressed. Messenger RNA (mRNA) and protein expression of DBH were studied throughout the estrous cycle. Additionally, DBH protein expression was examined in cultured mid luteal cells after tumour necrosis factor alpha/interferon gamma (TNFa/IFNg)-induced apoptosis or after treatment with NGF, IGF1, and TGFb1. DBH mRNA and protein expressions were detected throughout the cycle without significant changes in the protein level while mRNA showed a decrease at the developing stage (P < 0.05). Interestingly, NGF, IGF1, and TGFb1 increased DBH expression in cultured luteal cells (P < 0.05). The overall findings suggest non-neural source of noradrenaline in the bovine CL which appears to be regulated by NGF, IGF1, and TGFb1 indicating intraluteal molecules play an important and unrecognized role in the CL function

SPACA3gene variants in a New Zealand cohort of infertile and fertile couples

SPRASA (also referred to as SLLP1) is a protein identified in the acrosome of human sperm and encoded by the gene SPACA3. SPRASA is associated with sperm-oocyte recognition and binding, and may play a role in fertility. In order to determine whether variants in the SPACA3 gene are associated with human infertility, we undertook a genetic analysis of 102 infertile and 104 fertile couples. Three gene variants were identified using PCR-based DNA sequencing; 1) an insertion of TGC within a quadruple tri-nucleotide (TGC) repeat region in the 5’ untranslated region (UTR) (g.–22TGC(4_5), 2) a guanine to adenosine transition at position 239 (c.239G> A) resulting in a non-synonymous amino acid substitution from cysteine to tyrosine (p.C80Y) at position 80 in the putative transmembrane region, and 3) a novel nucleotide variant (c.691G> C) located in the 3’UTR. A functional effect of the g.–22TGC (4_5) was confirmed by a luciferase expression assay, while the effects of the variants c.239G> A and c.691G> C were predicted using in silico analysis. Although the frequencies of these variants were not significantly different between the infertile and fertile populations, we present evidence that the variants could affect the expression levels or function of SPRASA, thereby affecting a couple's fertility. Larger populations, especially individuals/couples with unexplained infertility, need to be screened for these variants to validate a relationship with fertility