Search Results (17)

In most mammals, a withdrawal of the pro-gestational hormone progesterone (P4) is necessary for labor onset. In murine cervix, P4 withdrawal is mediated by enzymes steroid 5-alpha-reductase type 1 (SRD5A1) and 20-alpha-hydroxysteroid-dehydrogenase (20α-HSD). Previously, we have shown that inflammatory stimuli induce 20α-HSD levels in uterine muscle (myometrium). Here, we hypothesized that (1) infectious inflammation alters the levels of both P4-metabolizing enzymes in mouse cervix, which consequently ceases P4-mediated inhibition of cervical remodeling, thereby inducing preterm labor (PTL); (2) a progestin, selective progesterone receptor modulator promegestone (aka R5020), non-metabolizable by 20α-HSD, can block lipopolysaccharide (LPS)-induced PTL in mice by maintaining P4 signaling and preventing cervical remodeling. Using RT-PCR and IHC/IF methods, we evaluated the effect of inflammation on the expression of both enzymes in mouse cervix and determined if R5020 can prevent cervical remodeling and PTL in mice. We found significant induction of SRD5A1 and 20α-HSD proteins (p < 0.01), as well as transcript levels of pro-inflammatory cytokines Il1b, Il6, chemokines Cxcl1, Ccl2, cervical ripening enzyme Has2, hyaluronic acid binding protein/HABP (p < 0.05), and a simultaneous decrease in major extracellular fibrillar proteins, collagen type 1 and type 3 (col1a1, col3a1), in mouse cervix during PTL. The prophylactic administration of R5020 in pregnant mice significantly inhibited cervical remodeling and prevented PTL irrespective of the route of LPS-induction, systemic or local. We concluded that R5020 is a promising novel drug application for preterm birth prevention. Full article

To improve the efficiency of prostaglandin F_2α (PG)-based estrus synchronization in sheep, this study assessed the effect of the gonadotropin-releasing hormone (GnRH) included in the long-interval PG treatment regimen for sheep estrus synchronization during the breeding season. In experiment 1, 30 multiparous Mongolian sheep (3–4.5 years old) were randomly divided into three groups. In the progesterone (P4)–equine chorionic gonadotropin (eCG) group (P4-eCG, n = 10), the ewes were synchronized with intravaginal P4 sponges for 14 days, and received an injection of 330 IU of eCG at sponge withdrawal. In the PG group (n = 10), the ewes received two doses of 0.1 mg PG with a 14 day interval. In the PG-GnRH-PG group (n = 10), the ewes were synchronized by two doses of 0.1 mg PG with a 14 day interval like that in the PG group, but received 50 μg of GnRH 7 days after the first injection of PG. It was found that, at the end of treatment, the number of corpus luteum on the ovaries and the concentration of the serum P4 in the PG-GnRH-PG group were significantly higher than that of the PG and P4-eCG groups. In experiment 2, 59 multiparous Mongolian sheep (3–4.5 years old) were assigned to three groups, like in experiment 1 (n = 20, 20 and 19 for the P4-eCG, PG and PG-GnRH-PG groups, respectively). The estrus of ewes in the PG-GnRH-PG group was more synchronous compared to the PG group. After insemination of the estrus ewes, the pregnancy rate was numerically but not significantly higher in the PG-GnRH-PG group than that in the P4-eCG and the PG groups. In a field test, 285 multiparous Hu sheep (3–4.5 years old) were randomly assigned to a P4-eCG group (n = 142) and PG-GnRH-PG group (n = 143). Timed artificial insemination showed no significant differences in the rates of pregnancy and lambing between the PG-GnRH-PG and P4-eCG groups. We conclude that the addition of GnRH in the long-interval PG protocol may improve the efficiency of PG-based estrus synchronization, and would represent a potential alternative to the conventional P4-eCG based protocol during the breeding season in sheep. Full article

There are fewer investigations conducted on human primary endometrial epithelial cells (HPEECs) compared to human primary endometrial stromal cells (HPESCs). One of the main reasons is the scarcity of protocols enabling prolonged epithelial cell culture. Even though it is possible to culture HPEECs in 3D over a longer period of time, it is technically demanding. In this study, we successfully established a highly pure, stable, and long-term viable human conditionally reprogrammed endometrial epithelial cell line, designated as eCRC560. These cells stained positive for epithelial markers, estrogen and progesterone receptors, and epithelial cell–cell contacts but negative for stromal and endothelial cell markers. Estradiol (ES) reduced the abundance of ZO-1 in a time- and dose-dependent manner, in contrast to the dose-dependent increase with the progestin dienogest (DNG) when co-cultured with HPESCs. Moreover, ES significantly increased cell viability, cell migration, and invasion of the eCRC560 cells; all these effects were inhibited by pretreatment with DNG. DNG withdrawal led to a significantly disrupted monolayer of eCRC560 cells in co-culture with HPESCs, yet it markedly increased the adhesion of eCRC560 to the human mesothelial MeT-5A cells. The long-term viable eCRC560 cells are suitable for in vitro analysis of HPEECs to study the epithelial compartment of the human endometrium and endometrial pathologies. Full article

This study deals with the immunological quality of canine colostrum in relation to the different parturition modes in bitches. It included four groups of bitches, who underwent vaginal parturition (VP), emergency cesarean section (EM-CS), elective cesarean section (EL-CS), or elective cesarean section with aglepristone injection 24 h prior to surgery (EL-A). Colostrum samples from 40 bitches of 18 breeds were taken immediately after the birth of the first puppy or directly after surgery, and four hours later. The concentrations of immunoglobulin G (IgG) and IgA were measured using ELISA tests. The initial IgG concentration was 18.3 ± 10.2 g/L, and the IgA concentration was 13.7 ± 5.8 g/L, respectively. Significantly lower IgG and IgA concentrations were observed in the EL-CS group compared to other groups. The administration of aglepristone led to an increase in IgG and IgA concentrations. Despite the lower immunoglobulin levels with EL-CS, the survival rate of the puppies was not affected. The study shows that immunoglobulin concentrations in colostrum vary between bitches, with the type of birth significantly influencing the levels. Progesterone withdrawal before birth could play a decisive role in increasing IgG and IgA concentrations in the mammary glands of the bitches. Full article

Premature regression of corpora lutea (PRCL) may adversely affect the outcome of hormonal ovarian superstimulation in small ruminants, and the total dose of exogenous gonadotropins used may be one of the causes of this condition. There were two major objectives of the present study: (1) to evaluate the effects of different superovulatory doses of porcine follicle-stimulating hormone (pFSH) on the biometry, blood perfusion (Doppler), and echotextural characteristics of luteal structures; and, (2) to determine the usefulness of biometric, vascular, and echotextural luteal variables, as well as measurements of circulating progesterone (P₄) concentrations for early detection of PRCL in superovulated Santa Inês ewes. Twenty-seven Santa Inês ewes received an intravaginal P₄-releasing device (CIDR) from Days 0 to 8 (Day 0 = random day of the anovulatory period). An IM injection of d-cloprostenol (37.5 μg) was given at the time of the CIDR insertion and withdrawal. On Day 6, all the ewes received 300 IU of eCG IM and were divided into three treatment groups (each n = 9): G100 (100 mg); G133 (133 mg); and G200 (200 mg of pFSH) administered IM every 12 h in eight injections. Transrectal ovarian ultrasonography and jugular blood sampling for serum P₄ measurements were performed on Days 11 to 15. On the day of embryo recovery (Day 15), all the ewes underwent diagnostic videolaparoscopy and were classified, based on their luteal characteristics, into three response groups: nCL (ewes with normal CL only); rCL (ewes with regressing CL only); and ewes with both nCL and rCL following the superovulatory regimen. Our present results indicate that the total pFSH doses of 100 mg and 200 mg result in similar ovulatory responses and luteal function/biometrics, although the percentage of donor ewes with nCL was greater (p < 0.05) for G100 compared with the G200 animals. An application of 133 mg of pFSH was associated with diminished luteogenesis. Lastly, circulating P₄ concentrations, ultrasonographic estimates of total luteal area, and CL pixel heterogeneity (standard deviation of numerical pixel values) are promising markers of luteal inadequacy in superovulated ewes. Full article

The aim of the present study was to assess whether the strategic supplementation of bypass LO can enhance reproductive indexes—fertility, lambing rate, and prolificacy—in dairy Sarda ewes at the end of lactation. To assess whether LO supplementation leads to the adsorptions of PUFAs and their subsequent utilization by the body tissues, milk composition and fatty acid content were analyzed. Forty-eight ewes were assigned to the following groups: the control group (CT; N = 24), fed with a control diet without LO; and the treatment group (LO; N = 24), fed with a diet supplemented with LO (10.8 g/ewe/day). Both diets had similar crude protein and energy levels and were offered for 38 days (−21 to +17 days after artificial insemination). The trial included an adaptation period (7 days) followed by a regular supplementation (31 days) period. Estrus synchronization was induced in all the ewes using an intravaginal sponge and equine chorionic gonadotropin. Fifty-five hours after pessaries withdrawal, all ewes were inseminated using the cervical route and fresh semen. Cholesterol (p < 0.01), high-density lipoprotein (p < 0.001), and triglyceride (p < 0.05) levels in plasma were higher in the LO group. Plasmatic levels of non-esterified fatty acids were lower in the LO group after the end of the supplementation period (p < 0.05). Milk unsaturated fatty acids (UFAs), monounsaturated fatty acids (MUFAs), total polyunsaturated fatty acids (PUFAs), PUFAs omega 3 (PUFAs-ω3) and 6 (PUFAs-ω6), and trans fatty acids were higher in the LO group (p < 0.001), while saturated fatty acids (SFAs) were higher in the CT group during the supplementation period (p < 0.001). Three days after the end of the supplementation period, the content of milk UFAs (p < 0.05), PUFAs (p < 0.001), MUFAs, and PUFAs-ω6 (p < 0.01) were still higher in the LO group. whereas SFA was higher in the CT group (p < 0.01). There was no difference between groups in terms of ovulation rate, progesterone levels in plasma, fertility rate, prolificacy, and total reproductive wastage. However, the total area of luteal tissue was higher in the LO group (p < 0.01). Results obtained demonstrated that LO supplementation exerts a positive role in corpus luteum size at the onset of the peri-implantation period in Sarda dairy ewes. Additionally, the results obtained in the present study showed that the use of dietary bypass LO affects lipid metabolites in plasma and milk fatty acid profiles, demonstrating the ALA uptake by body tissues. Full article

Sexual activity in domestic goats is positively influenced by reducing the photoperiod. Various protocols have therefore been developed in goats for the induction and synchronization of estrus during those months in which their sexual activity is reduced. The present observational study evaluates the periovulatory hormonal profile in Payoya goats (n = 24), during a non-favorable photoperiod (i.e., spring), being treated for estrus induction. The treatment comprised the vaginal insertion of sponges impregnated with progestogen (fluorogestone acetate, FGA), together with cloprostenol and equine chorionic gonadotrophin (eCG), 48 h before the end of the treatment. When the treatment ended, the plasma concentrations of the LH, FSH, progesterone and estradiol were determined. The goats were inseminated 46 h after the sponge withdrawal, and a pregnancy diagnosis was carried out 40–45 days after the insemination. Various parameters were monitored, such as the peaks of luteinizing hormone (LH), follicle-stimulating hormone (FSH) and estradiol, and their respective intervals, in reference to the time of the sponge withdrawal. The conception rate was 62.5%, and the kidding rate was 50%. The results record the hormonal release pattern after the estrus synchronization treatment based on the FGA, and the differences between the pregnant and non-pregnant goats. The findings suggest that the LH peak produced after the estrus synchronization treatment, both in terms of the amplitude and the time of increment, is involved in the reproductive failure detected. Full article

Background: Social behavior is mediated by steroid hormones, whereby various lines of evidence indicate that metformin might improve the symptoms of social withdrawal. This directly yields to the aim of the study to correlate the impact of metformin treatment on the potential alterations in steroid hormone homeostasis, which is ultimately impacting social behavior. Therefore, urinary samples of patients before and after treatment with metformin will be correlated to social behavior to elucidate potential changes in steroid hormone profiles and social behavior. Material and Methods: An observational study in healthy adults with a new indication for metformin. Steroid hormone analysis, including the most prominent androgen, estrogen, progesterone, aldosterone, corticosterone, cortisone and cortisol metabolites analyzed with gas chromatography–mass spectrometry and a questionnaire on social behavior (Autism Spectrum Questionnaire (AQ)) will be administered prior to and after around a 12-week phase of metformin treatment. Discussion: It is likely that due to different pathophysiological mechanisms such as an effect on the respiratory chain in mitochondria or via AMP-activated protein kinase, a general alteration of steroid hormone levels can be detected prior to post treatment. The encompassing measurement of steroid hormones shall give hints concerning the involvement of specific cascades yielding potential pharmacological targets for future research. Full article

Stress susceptibility could play a role in developing premenstrual anxiety due to abnormalities in the hypothalamus–pituitary–adrenal (HPA) axis and impairments in the GABAA receptors’ benzodiazepine (BDZ) site. Hence, we studied the stress-vulnerable Wistar Kyoto rat strain (WKY) to evaluate progesterone withdrawal (PW) effects on anxiety, HPA axis response, and to explore indicators of GABAA functionality in the BDZ site. For five days, ovariectomized WKY rats were administered 2.0 mg/kg of progesterone. Twenty-four hours after the last administration, rats were tested in the anxiety-like burying behavior test (BBT) or elevated plus maze test (EPM), and corticosterone was determined. [3H]Flunitrazepam binding autoradiography served as the BDZ binding site index of the GABAA receptor in amygdala nuclei and hippocampus’s dentate gyrus (DG). Finally, different doses of diazepam in PW-WKY rats were tested in the BBT. PW induced anxiety-like behaviors in both BBT and EPM compared with No-PW rats. PW increased corticosterone, but was blunted when combined with PW and BBT. PW increased [3H]Flunitrazepam binding in the DG and central amygdala compared with No-PW rats. Diazepam at a low dose induced an anxiogenic-like response in PW rats, suggesting a paradoxical response to benzodiazepines. Overall, PW induced anxiety-like behavior, a blunted HPA axis response, and higher GABAAR/BZD binding site sensitivity in a stress-vulnerable rat strain. These findings demonstrate the role of stress-susceptibility in GABAAR functionality in a preclinical approximation of PMDD. Full article

Asthma is a frequent medical condition in adolescence. The worsening of the most common symptoms perimenstrually is defined as perimenstrual asthma (PMA). The cause of PMA remains unclear, but a role for hormonal milieu is plausible. Data on PMA in adolescents are limited, and its management is not fully established. We aimed to discuss the PMA phenomenon in young females from pathophysiology to preventive strategies, focusing on the relationship with the hormonal pattern. The fluctuation of estrogens at ovulation and before menstruation and the progesterone secretion during the luteal phase and its subsequent withdrawal seem to be the culprits, because the deterioration of asthma is cyclical during the luteal phase and/or during the first days of the menstrual cycle. Conventional asthma therapies are not always effective for PMA. Preventive strategies may include innovative hormonal contraception. Even a possible beneficial effect of other hormonal treatments, including estrogens, progestogens, and androgens, as well as leukotriene receptor antagonists and explorative approach using microbial-directed therapy, is considered. The underlying mechanisms, through which sex-hormone fluctuations influence asthma symptoms, represent a challenge in the clinical management of such a distressing condition. Further studies focused on young females are mandatory to promote adolescent health. Full article

Bleeding in newborns and young girls fascinated writers for more than a millennium. Initially, there was confusion between neonatal bleeding, early menstruation due to precocious puberty, and hemorrhage due to disease. During the 19th century descriptions appeared of what is referred to today as ‘neonatal menstruation’ or ‘neonatal uterine bleeding’. By the turn of the century, Halban linked bleeding to active substances present during pregnancy and hypothesized that, while the maternal uterus reacts with decidua formation, the “weaker” fetal uterus reacts only with menstrual-like changes. Despite this clear description, several alternative theories endured for decades. Bleeding was believed to be due to a ‘catarrhal’ or neoplastic state of the genital tract, pulmonary circulatory disorder, congenital heart malformations, closure of the umbilical cord or affections of the intestine. During the 1950s, progesterone response and resistance were proposed to explain the pathogenesis of bleeding and its low incidence. The fetal endometrium is resistant to the high circulating progesterone. A decidual response is infrequent and results in menstrual shedding upon progesterone withdrawal after birth. Further research linked fetal stress consequent to pregnancy complications and post-maturity to increased incidence and preterm birth to reduced incidence of neonatal uterine bleeding. Full article

Background and Objectives: Therapeutic interventions targeting molecular factors involved in the transition from uterine quiescence to overt labour are not substantially reducing the rate of spontaneous preterm labour. The identification of novel rational therapeutic targets are essential to prevent the most common cause of neonatal mortality. Based on our previous work showing that Tbx2 (T-Box transcription factor 2) is a putative upstream regulator preceding progesterone withdrawal in mouse myometrium, we now investigate the role of TBX2 in human myometrium. Materials and Methods: RNA microarray analysis of (A) preterm human myometrium samples and (B) myometrial cells overexpressing TBX2 in vitro, combined with subsequent analysis of the two publicly available datasets of (C) Chan et al. and (D) Sharp et al. The effect of TBX2 overexpression on cytokines/chemokines secreted to the myometrium cell culture medium were determined by Luminex assay. Results: Analysis shows that overexpression of TBX2 in myometrial cells results in downregulation of TNFα- and interferon signalling. This downregulation is consistent with the decreased expression of cytokines and chemokines of which a subset has been previously associated with the inflammatory pathways relevant for human labour. In contrast, CXCL5 (C-X-C motif chemokine ligand 5), CCL21 and IL-6 (Interleukin 6), previously reported in relation to parturition, do not seem to be under TBX2 control. The combined bioinformatical analysis of the four mRNA datasets identifies a subset of upstream regulators common to both preterm and term labour under control of TBX2. Surprisingly, TBX2 mRNA levels are increased in preterm contractile myometrium. Conclusions: We identified a subset of upstream regulators common to both preterm and term labour that are activated in labour and repressed by TBX2. The increased TBX2 mRNA expression in myometrium collected during a preterm caesarean section while in spontaneous preterm labour compared to tissue harvested during iatrogenic preterm delivery does not fit the bioinformatical model. We can only explain this by speculating that the in vivo activity of TBX2 in human myometrium depends not only on the TBX2 expression levels but also on levels of the accessory proteins necessary for TBX2 activity. Full article

Estrus synchronization of gilts can be achieved by feeding the orally active progestogen altrenogest (AT) to cycling gilts at 20 mg/day for 14 to 18 days with gilts usually returning to estrus 4 to 8 days after the last feeding. In practice, gilts failing to exhibit estrus by 6 days after AT withdrawal may compromise weekly breeding targets. The cause of prolonged intervals to estrus are unknown but may involve prolonged suppression due to the release of progesterone (P4), and by extension AT, from adipose tissues. The present study examined relationships between gilt P2 backfat depth (<13.5 mm, 14–16.5 mm, >17 mm groups), the AT withdrawal to estrus interval, and subsequent reproductive performance in gilts. We noted longer intervals to estrus in gilts with greater P2 backfat depths (p < 0.0001), and higher serum P4 concentrations on the last day of AT feeding and at estrus detection (p < 0.05). Total born litter sizes were unaffected by backfat depth, but pigs born alive progressively decreased with increasing backfat depth with the fattest gilts producing the fewest liveborn pigs (p < 0.05). Taken together, these data suggest that adipose tissues may provide a reservoir of steroid, with its release from fatter gilts having potential negative effects on their subsequent reproductive performance. Full article

Oral contraceptives (OCs) containing estrogen and progesterone analogues are widely used amongst reproductive-aged women, but their neurocognitive impact is poorly understood. Preliminary studies suggest that OCs improve verbal memory and that OCs with greater androgenic activity may improve visuospatial ability. We sought to explore the cognitive impact of OCs by assessing performance of OC users at different stages of the OC cycle, and comparing this performance between users of different OC formulations according to known androgenic activity. We conducted a prospective, observational trial of OC users, evaluating cognitive performance with CogState software on two occasions: days 7–10 of active hormonal pill phase, and days 3–5 of the inactive pill phase (coinciding with the withdrawal bleed resembling menstruation). Thirty-five OC users (18 taking androgenic formulations, 17 taking anti-androgenic) were assessed. Analysis by androgenic activity showed superior performance by users of androgenic OCs, as compared to anti-androgenic OCs, in visuospatial ability and facial affect discrimination tasks. A growing understanding of cognitive effects of OC progestin androgenicity may have implications in choice of OC formulation for individuals and in future OC development. Full article

Granulosa cells (GCs) are the key components of ovarian follicles for regulating oocyte maturation. Previous established GC lines have allowed prolonged proliferation, but lost some physiological features owing to long-term immortalization. This study was to establish an induced immortal porcine GC line with reversible proliferation status by the tetracycline inducible (Tet-on) 3G system. Our conditional immortal porcine GCs (CIPGCs) line steadily propagated for at least six months and displayed primary GC morphology when cultured in the presence of 50 ng/mL doxycycline [Dox (+)]. Upon Dox withdrawal [Dox (–)], Large T-antigen expression, reflected by mCherry fluorescence, gradually became undetectable within 48 h, accompanied by less proliferation and size increase. The levels of estradiol and progesterone, and the expression of genes associated with steroid production, such as CYP11A1 (cytochrome P450 family 11), 3β-HSD (3β-hydroxysteroid dehydrogenase), StAR (steroidogenic acute regulatory protein), and CYP19A1 (cytochrome P450 family 19 subfamily a member 1), were all significantly higher in the Dox (–) group than Dox (+) group. The CIPGCs could switch into a proliferative state upon Dox induction. Interestingly, the expression of StAR and CYP19A1 in the CIPGCs (–Dox) was significantly increased by adding porcine follicular fluid (PFF) to mimic an ovary follicle environment. Moreover, PFF priming the CIPGCs in Dox (–) group resulted in similar estradiol production as that of primary GC, and enabled this cell line to respond to gonadotrophins in estradiol production. Collectively, we have established an inducible immortal porcine GC line, which offers a unique and valuable model for future research on the regulation of ovarian functions. Full article