Human Chorionic Gonadotropin Hormone Research Articles

Use of kisspeptin to trigger oocyte maturation during in vitro fertilisation (IVF) treatment.

Infertility is a major global health issue and is associated with significant psychological distress for afflicted couples. In vitro fertilisation (IVF) utilises supra-physiological doses of stimulatory hormones to induce the growth of multiple ovarian follicles to enable surgical retrieval of several oocytes for subsequent fertilisation and implantation into the maternal endometrium. The supra-physiological degree of ovarian stimulation can lead to potential risks during IVF treatment, including ovarian hyperstimulation syndrome (OHSS) and multiple pregnancy. The choice of oocyte maturation trigger, such as human chorionic gonadotrophin (hCG) or gonadotrophin releasing hormone agonist (GnRHa), can impact both the efficacy of IVF treatment with a bearing on luteal phase hormonal dynamics and thus the degree of luteal phase support required to maintain optimal pregnancy rates, as well as on safety of treatment with particular respect to the risk of OHSS. Kisspeptin regulates gonadotrophin releasing hormone (GnRH) release and is therefore a key regulator of the hypothalamo-pituitary-gonadal (HPG) axis. Kisspeptin has been shown to be requisite for the occurrence of the physiological ovulatory luteinising hormone (LH) surge. In this review, we discuss the potential use of kisspeptin as a novel trigger of oocyte maturation.

Prevalence of high affinity naturally occurring IgG2 antibodies against human chorionic gonadotropin and its subunits in patients with ovarian cyst

Naturally occurring antibodies to tumour antigens are gaining interest as clinically important cancer biomarkers for early diagnosis, prognosis and for the development of anti-cancer therapeutics. The glycoprotein αβ heterodimer hormone human chorionic gonadotropin (hCG) and its β subunit (hCGβ) are produced by various cancers, and their increased serum levels correlate with poor prognosis. We have previously reported that patients with benign ovarian cysts, but not the malignant tumours, were characterized by augmented serum levels of naturally-occurring IgG antibodies to hCG and hCGβ. Here we further characterise these antibodies in patients with ovarian cysts.IgG and IgM antibody binding to whole hCG, hCGβ, hCGα, hCGβ C-terminal peptide (hCGβCTP), and the hCGβ core fragment (hCGβCF) were measured in the sera from 36 patients with ovarian cysts and 12 healthy non-pregnant women using a standard ELISA. IgG subclass usage and affinity was also determined together with cross-binding to whole hCG and its subunits of four selected commercial monoclonal antibodies generated against ovarian cyst mucins.Our results showed that 91.7% of the sera tested contained elevated IgG, but not IgM antibodies to one or several antigens, with an overwhelming prevalence of high affinity IgG2 indicating their binding to carbohydrate epitopes and possibly ovarian cyst mucins. Anti-mucin commercial antibody ab212418 (Abcam) produced against Gal1-3GalNAc, exhibited strong cross-binding to hCGαβ, hCGβ, hCGα and hCGβCTP. The protective anti-cancer potential of these antibodies will be further investigated and could lead to the development of novel treatment strategies for ovarian cancer.

Diagnosis of Cornual Pregnancy in the Emergency Room: a Rare Case Report

Introduction: Ectopic or extrauterine pregnancy is a life-threatening medical condition in which a fertilized egg is implanted elsewhere than in the uterine cavity, commonly in the fallopian tubes. However, it can also occur in the peritoneal cavity, ovaries, and cervix. There are several etiological factors contributing to ectopic pregnancies, such as a history of pelvic inflammatory diseases, habitual abortions, and miscarriages, previous ectopic pregnancies, fertility procedures, as well as increased maternal age that has been correlated with a higher risk of carrying an ectopic pregnancy. Often, the first alerting sign in any ectopic pregnancy is a pain in the pelvic region, along with vaginal bleeding usually spotted between the 6th and 11th gestational week. Other symptoms include vomiting, nausea, tachycardia, and in severe cases hypovolemic shock due to internal bleeding induced by the rupture of the tubes. The key to diagnosing an ectopic pregnancy is transvaginal ultrasound, alongside the measurement of beta human chorionic gonadotropin hormone (beta-hCG) concentrations, which are usually found to be low in such cases. The management of ectopic pregnancy is either medical or surgical. Case report: We are reporting the case of a 32-year-old pregnant woman, who presented to the emergency department for diffuse abdominal pain and recurrent episodes of vomiting and diarrhea of one-week duration. The patient was stable on admission, but later on, she showed a rapid shift in her vital signs. Blood tests were obtained, and pelvic ultrasound was done, followed by a Computed Tomography (CT) scan that revealed the presence of a right cornual ectopic pregnancy. A rapid surgical intervention was carried out to save the patient’s life and alleviate the pain. Conclusion: Ectopic pregnancy is an urgent condition that needs to be treated immediately to avoid its fatal consequences. Several risk factors stand behind it, and a usual history and physical gynecological examinations are insufficient to detect it, hormone screening tests additionally to ultrasounds are highly recommended to decrease the incidence of mortality. A proper follow-up with a specialist could avoid the risk of an ectopic pregnancy as well as its undesirable outcomes.

Comparison of an HCG-only trigger versus dual trigger for final oocyte maturation in a progestin-primed ovarian stimulation protocol

Research questionIs there any difference in clinical outcomes between a human chorionic gonadotrophin (HCG)-only trigger and a dual trigger combining gonadotrophin-releasing hormone agonist (GnRHa) and HCG in a progestin-primed ovarian stimulation (PPOS) protocol? DesignThis retrospective cohort study included women younger than 40 years old with a normal ovarian reserve who underwent IVF/intracytoplasmic sperm injection treatment with a PPOS protocol. Participants were allocated to two groups according to the triggering medicines. The clinical outcomes were compared, with cumulative live birth rate (CLBR) being the primary outcome. ResultsIn total, 1066 women were included, 565 in the HCG-only group and 501 in the dual trigger group. Demographic parameters were comparable between the groups. Fewer oocytes were retrieved in the HCG-only trigger group (dual trigger 12.56 ± 7.12 versus HCG-only trigger 11.62 ± 6.02, P = 0.020). No significant difference was observed in the numbers of two-pronuclear embryos (7.12 ± 4.90 versus 6.76 ± 4.45, P = 0.208) and high-quality embryos (4.01 ± 3.70 versus 3.96 ± 3.32, P = 0.815). The CLBR after one complete cycle was also similar (40.72% versus 43.72%, P = 0.354). Multivariate logistic analysis confirmed that the trigger method had no association with CLBR (odds ratio [OR] 0.763, 95% confidence interval [CI] 0.578–1.005, P = 0.055) in the PPOS-treated patients. ConclusionsCompared with the HCG-only trigger group, comparable embryological and clinical outcomes were achieved, although more oocytes were retrieved in the dual trigger group. This suggests that there may be no extra benefit from dual triggering, and that it should not be recommended for routine use in the general population undergoing PPOS protocols.

Evaluation of hormonal protocols for induction of synchronized estrus on reproductive indices in Kangal-Akkaraman ewes during the outbreeding season

The objective of this study was to assess the effect of human chorionic gonadotropin (hCG) and gonadotropin-releasing hormone (GnRH) treatment before sponge insertion and anti-Müllerian hormone (AMH) concentration in ewes which were either pregnant or non-pregnant during the previous breeding season. In the study, 375 (3–6-year-old) Kangal-Akkaraman ewes that conceived or did not conceive in the preceding breeding season, and 36 rams (3–6-year-old) were used. The sheep were randomly divided into 5 equal groups. Blood samples were taken before stimulation (day −4), the day stimulation was initiated (day 0), after stimulation (day 9), on the mating day (days 11–12). AMH concentrations were measured in the samples taken on − 4th day. As a conventional method, a sponge containing 20 mg of flugeston acetate (Chronogest® CR, MSD, Turkey) was placed intravaginally (day 0) into to study ewes. The ewes were intramuscularly administered 131.5 µg prostaglandin F2α (Estrumate, MSD, Turkey) and 600 IU equine chorionic gonadotropin (Chronogest® PMSG 6000, MSD, Turkey) simultaneously with sponge removal 9 days after insertion. No other treatment was applied to group 5 which served as control. Ewes in groups 1, 2, and 3 received an intramuscular injection of GnRH (4.2 µg of Buserelin acetate, Receptal, MSD, Turkey) at the time of insertion of progesterone sponge. GnRH or 600 IU hCG (Chorulon®, MSD, UK) was administered 4 days prior to sponge insertion for the ewes in the groups 2, and 3, respectively. As for the group 5 group, hCG (instead of GnRH) was administered to the ewes at the beginning of the progesterone treatment. Ewes in estrus were detected with a teaser ram and taken into a different pen with rams. Transrectal ultrasonography (Mindray DP50/Vet/US, rectal probe) was achieved during days the 30–35 following mating. Transabdominal ultrasonography was achieved during days 60–65 to detect late embryonic and early fetal death. The estrus rate was 71.19 % out of breeding season for all ewes. The pregnancy rate on day 30 post-mating was 32 % in the control group and was 31.00 %. No significant differences were observed between groups for pregnancy rates on day 30. The mean pregnancy rate in sheep that conceived and did not conceived in the preceding breeding season of the study was 32.03 % and 32.06 % respectively (P > 0.05). After treatments, AMH concentrations in non-pregnant were significantly higher than pregnant ewes (P < 0.005). In conclusion, although the AMH concentration is significantly higher in non-pregnant ewes, the AMH concentration may not be a suitable biomarker to predict induction success outside breeding season. Progesterone-based protocols may achieve an acceptable reproductive outcome in lactating ewes and the ones which experienced infertility.

Reproductive outcomes of dual trigger with combination GnRH agonist and hCG versus trigger with hCG alone in women undergoing IVF/ICSI cycles: a retrospective cohort study with propensity score matching

BackgroundDespite a large number of studies on the selection of trigger drugs, it remains unclear whether the dual trigger with human chorionic gonadotropin (hCG) and gonadotropin-releasing hormone (GnRH) agonist, compared to the trigger with hCG alone, can improve the reproductive outcome of patients undergoing assisted reproductive technology. Therefore, this study aimed to compare the laboratory and clinical outcomes of dual trigger versus single trigger.MethodsIn this retrospective cohort study, we evaluated 520 in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) cycles between July 2014 and September 2020 at the Reproductive and Genetic Center of Integrative Medicine, The Affiliated Hospital of Shandong University of Traditional Chinese Medicine. All patients underwent IVF/ICSI treatment with fresh embryo transfer using the GnRH antagonist protocol. We used propensity score matching to control for confounding variables and binary logistic regression analysis to determine the correlations between trigger methods and pregnancy outcomes. After propensity score matching, 57 cycles from each group were evaluated and compared for laboratory or clinical outcomes in this retrospective cohort study.ResultsThere was no significant difference in the number of oocytes retrieved, embryos available, top-quality embryos, or the rate of normal fertilization between the dual-trigger and single-trigger protocols, respectively. The incidence of ovarian hyperstimulation syndrome, implantation rate, biochemical pregnancy rate, clinical pregnancy rate, ectopic pregnancy rate, early miscarriage rate, and live birth rate were also similar between the two groups, while the miscarriage rate (37.0% vs. 12.5%, p = 0.045) was higher in the dual-trigger than the single-trigger group. Subsequent binary logistic regression analysis showed that age was a remarkably significant independent predictor of both clinical pregnancy rate (odds ratio = 0.90, 95% confidence interval: 0.84–0.97, p = 0.006) and live birth rate (odds ratio = 0.89, 95% confidence interval: 0.82–0.97, p = 0.005).ConclusionsTherefore, dual-trigger for final oocyte maturation might increase miscarriage rate, but in terms of the laboratory and other pregnancy outcomes such as clinical pregnancy rate, early miscarriage rate or live birth rate, there was no evidence to show that dual trigger was superior to an hCG-trigger alone for patients undergoing GnRH-antagonist cycles with fresh embryo transfer.Trial registrationRetrospectively registered.

Study on the Influencing Mechanism of Human Chorionic Gonadotropin (hCG) on Oocyte Maturation in Patients with Polycystic Ovary Syndrome.

The study was aimed at investigating the influence of human chorionic gonadotropin (hCG) hormone on oocyte maturation in the patients with polycystic ovary syndrome (PCOS). A total of 54 patients with PCOS who received in vitro maturation (IVM) treatment in the Cheeloo College of Medicine, Shandong University, were divided into two groups: one group who underwent hCG injections was the observation group (OG; n = 27) and other was the control group (CG; n = 27) with no hCG injection. The oocyte development and the expression of steroid hormone synthesis-related genes including gonadotropin-releasing hormone receptor (GnRHR), Conexin43, epidermal growth factor-related genes, luteinizing hormone/choriogonadotropin receptor (LHCGR), epiregulin (EREG), and vascular endothelial growth factor (VEGF) were examined. The human ovarian granulosa cell line (SVOG cells) and ovarian epithelial cell line (HOSEpiC cells) were employed to analyze the effect of hCG on the biological behaviour of cells. As a result, OG showed higher normal fertilization, cleavage, and high-qualified embryo rate than CG. Expression levels of GnRHR, Cx43, LHCGR, EREG, and VEGF were significantly elevated in granulosa cells in the OG group. Western blot revealed that phosphoinositide 3-kinase (PI3K), protein kinase B (AKT), and rapamycin (mTOR) proteins were decreased in granulosa cells under hCG intervention. A biological behaviour test indicated that the multiplication capacity of hCG-intervened SVOG and HOSEpiC was increased, while the apoptosis was decreased. In conclusion, hCG could accelerate follicular development and oocyte maturation by activating oocyte maturation genes in PCOS patients, which could significantly improve and popularize the application of IVM technology.

P-663 Dydrogesterone + progesterone gel vs. subcutaneous aqueous progesterone + progesterone gel for luteal phase supplementation in IVF cycles in women with previous IVF failure

Abstract Study question Does the live birth rate differ between the two regimens for luteal phase supplementation in IVF cycles in women with a history of IVF failure? Summary answer The live birth rate did not differ with dydrogesterone + progesterone gel vs. subcutaneous aqueous progesterone + progesterone gel. What is known already Supplementation of the luteal phase in in vitro fertilization (IVF) cycles is crucial to increase the chance of a live birth. Progesterone is the treatment of choice due to a lower risk of complications compared to human chorionic gonadotropin (hCG) and gonadotropin-releasing hormone (GnRH) analogues. There is no preferred progestogen for use in the general population of women undergoing IVF. The optimal progesterone treatment regimen in the event of previous IVF failure is unknown. Study design, size, duration A prospective randomized tertiary single-center study was conducted. The study included 161 women, aged 25-43 years, with FSH on cycle day 2-4 ≤10 IU/L, with at least one failed IVF cycle, undergoing government-funded IVF in the years 2015-2016. Participants/materials, setting, methods 161 included women were randomized after oocytes retrieval to 2 groups of luteal IVF cycle phase supplementation: dydrogesterone (Duphaston®) + progesterone in vaginal gel (Crinone®) or aqueous progesterone solution in subcutaneous injection (Prolutex®) + progesterone in vaginal gel (Crinone®). At least one blastocyst was obtained in the enrolled women. All women underwent fresh embryo transfer. After viable pregnancy confirmation, treatment was continued until the 12th week of gestation. Main results and the role of chance The indications for IVF were: male factor (51%), tubal factor (27%), endometriosis (24%), decreased ovarian reserve (17%), idiopathic infertility (14%), ovulation disorders (6%), occurring in various combinations. Out of 161 women enrolled in the study, 94 had a history of 1 failed IVF cycle, and 67 had at least 2 failed IVF cycles. Among 94 women with 1 failed IVF, 62 received dydrogesterone + progesterone gel vs. 32 aqueous progesterone + progesterone gel, and the percentages of live births, clinical pregnancies and biochemical pregnancies were: 29 vs. 21% (p = 0.37), 10 vs. 6% (p = 0.53), 5 vs. 12% (p = 0.21). Among 67 women with at least 2 failed IVFs, 24 received dydrogesterone + progesterone gel vs. 43 aqueous progesterone + progesterone gel, and the percentages of live births, clinical pregnancies and biochemical pregnancies were: 17 vs. 32% (p = 0.19), 4 vs. 0% (p = 0.14), 9 vs. 9% (p = 0.95). The results were comparable with both treatment regimens. There were no significant adverse events reported for both treatment regimens. Limitations, reasons for caution The limitations of the study are the small sample size, its single-center character and heterogeneity of indications for IVF. Wider implications of the findings In light of the lack of evidence of greater efficacy of either of the two luteal phase supplementation regimens of the IVF cycle in women with a history of IVF failure, the choice of treatment should be guided by other considerations, i.e. potential side effects, dosing convenience and patient preference. Trial registration number 1072.6120.47.2021

Triggering with 1,500 IU of human chorionic gonadotropin plus follicle-stimulating hormone compared to a standard human chorionic gonadotropin trigger dose for oocyte competence in in vitro fertilization cycles: a randomized, double-blinded, controlled noninferiority trial

To assess if triggering with 1,500 IU of human chorionic gonadotropin (hCG) with 450 IU of follicle-stimulating hormone (FSH) induces noninferior oocyte competence to a standard dose of hCG trigger used in invitro fertilization (IVF). The alternative trigger will be considered noninferior if it is at least 80% effective in promoting oocyte competence. Randomized, double-blinded, controlled noninferiority trial. Academic infertility practice. Women aged 18-41 undergoing IVF with antral follicle count ≥8, body mass index ≤30 kg/m2, and no history of ≥2 IVF cycles canceled for poor response were enrolled. Participants with a serum estradiol >5,000 pg/mL on the day of trigger were excluded because of high risk of ovarian hyperstimulation syndrome. Participants were randomized to receive an alternative trigger of 1,500 IU of hCG plus 450 IU of FSH or a standard trigger dose of hCG (5,000 or 10,000 IU) for final oocyte maturation. The primary outcome was total competent proportion, defined as the probability of 2 pronuclei from an oocyte retrieved. The alternative trigger will be considered noninferior to the standard trigger if a 1-sided 95% confidence interval (CI) of the relative risk (RR) is not <0.8. Secondary outcomes included oocyte recovery and maturity, intracytoplasmic sperm injection fertilization, embryo quality, pregnancy rates, as well as serum and follicular hormones. Secondary outcomes were compared using a 2-sided superiority test. Outcomes were analyzed by intention-to-treat and per-protocol. A total of 105 women undergoing IVF were randomized from May 2015 to June 2018. The probability of the primary outcome was 0.59 with the alternative trigger and 0.65 with the standard trigger, with a RR of 0.91 and a 1-sided 95% CI of 0.83. Noninferiority of the alternative trigger was demonstrated. Live birthrate from all fresh transfers in the alternative trigger group vs. standard trigger was 46.9 vs. 46.4% (RR, 1.01; 95% CI, 0.62-1.62), respectively. Live birthrate per randomized participant was 48.1% in the alternative trigger group vs. 62.7% with the standard trigger (RR, 0.73; 95% CI, 0.48-1.11). No participants had a failed retrieval. Triggering with 1,500 IU of hCG plus 450 IU of FSH promoted noninferior oocyte competence compared to a standard hCG trigger dose. NCT02310919.

The Role of Human Chorionic Gonadotropin Beta (hCGβ) in HPV-Positive and HPV-Negative Oropharyngeal Squamous Cell Carcinoma.

Simple SummaryOropharyngeal squamous cell carcinoma (OPSCC) is often associated with human papillomavirus (HPV). The management of OPSCC remains a challenge, and HPV-negative disease is commonly associated with impaired survival. Novel insights into diagnostics and treatment modalities are warranted to improve disease outcome. Various potential biomarkers affecting prognosis in OPSCC have been distinguished. Our aim was to study the role of the free β-subunit of human chorionic gonadotropin hormone in HPV-negative and HPV-positive OPSCC.Background: This study was carried out to observe the upregulation of the free β-subunit of human chorionic gonadotropin (hCGβ) and its prognostic significance in human papillomavirus (HPV)-positive and HPV-negative oropharyngeal squamous cell carcinoma (OPSCC). Materials and methods: A total of 90 patients with OPSCC treated with curative intent at the Helsinki University Hospital (HUS), Helsinki, Finland, during 2012–2016 were included. Serum samples were collected prospectively, and their hCGβ concentrations (S-hCGβ) were determined by an immunofluorometric assay. The expression of hCGβ in tumor tissues was defined by immunohistochemistry (IHC). HPV determination was performed by combining p16-INK4 IHC and HPV DNA PCR genotyping. Overall survival (OS) and disease-specific survival (DSS) were used as survival endpoints. Results: S-hCGβ positivity correlated with poor OS in the whole patient cohort (p < 0.001) and in patients with HPV-negative OPSCC (p < 0.001). A significant correlation was seen between S-hCGβ and poor DSS in the whole cohort (p < 0.001) and in patients with HPV-negative OPSCC (p = 0.007). In a multivariable analysis, S-hCGβ was associated with poor DSS. Of the clinical characteristics, higher cancer stage and grade were associated with S-hCGβ positivity. No statistically significant correlation with tissue positivity of hCGβ was seen in these analyses. Conclusion: S-hCGβ may be a potential independent factor indicating poor prognosis, notably in HPV-negative OPSCC.

Gambaran Berat Jenis Urin Dan Kadar Protein Urin Pada Ibu Hamil Di Upk Puskesmas Kampung Dalam Pontianak Timur

Laboratory examinations are carried out to determine diseases that can affect pregnancy, one of the tests carried out is routine urine. The presence of the hormone HCG (Human Chorionic Gonadotrophin) in the urine can be used for a pregnancy test. Kidney function, can be known through a protein test in the urine, the presence of protein in the urine leads to pre-eclampsia in pregnant women. High urine specific gravity can be caused by lack of fluid consumption, the presence of sugar in the urine, and protein in the urine (usually in people with kidney disorders) or certain medications. This study aims to describe the description of urine specific gravity and urine protein levels in pregnant women in the UPK Puskesmas Kampung Dalam, Pontianak Timur. This is a descriptive survey research using the accidental sampling technique. Measurements and observations of research variables were measurements of urine specific gravity and urine protein content which were carried out at the UPK Laboratory of the Kampung Dalam Puskesmas, Pontianak Timur. The sample used is a urine sample when pregnant women who meet predetermined criteria, the examination is carried out using a Urine Analyzer. Respondents of this study found 30 people. The results of 30 respondents pregnant women measuring urine specific gravity whose normal limit is 1,005-1,030 obtained urine specific gravity of 1,010 as many as 6 respondents (20%), 1,015 as many as 9 respondents (30%), 1,020 as many as 2 respondents (6,7% ). 1,025 as many as 6 respondents (20%), and 1,030 as many as 7 respondents (23.3%). While the results of the measurement of urine protein levels were positive 1 as many as 4 respondents (13.3%), positive 2 as many as 1 respondent (3.4%), and Negative as many as 25 respondents with a percentage (83.3%).

Expression and function of the luteinizing hormone choriogonadotropin receptor in human endometrial stromal cells

The human luteinising hormone choriogonadotropin receptor (LHCGR) is a G-protein coupled receptor activated by both human chorionic gonadotropin (hCG) and luteinizing hormone (LH), two structurally related gonadotropins with essential roles in ovulation and maintenance of the corpus luteum. LHCGR expression predominates in ovarian tissues where it elicits functional responses through cyclic adenosine mononucleotide (cAMP), Ca2+ and extracellular signal-regulated kinase (ERK) signalling. LHCGR expression has also been localized to the human endometrium, with purported roles in decidualization and implantation. However, these observations are contentious. In this investigation, transcripts encoding LHCGR were undetectable in bulk RNA sequencing datasets from whole cycling endometrial tissue and cultured human endometrial stromal cells (EnSC). However, analysis of single-cell RNA sequencing data revealed cell-to-cell transcriptional heterogeneity, and we identified a small subpopulation of stromal cells with detectable LHCGR transcripts. In HEK-293 cells expressing recombinant LHCGR, both hCG and LH elicited robust cAMP, Ca2+ and ERK signals that were absent in wild-type HEK-293 cells. However, none of these responses were recapitulated in primary EnSC cultures. In addition, proliferation, viability and decidual transformation of EnSC were refractory to both hCG and LH, irrespective of treatment to induce differentiation. Although we challenge the assertion that LHCGR is expressed at a functionally active level in the human endometrium, the discovery of a discrete subpopulation of EnSC that express LHCGR transcripts may plausibly account for the conflicting evidence in the literature.

Mechanistic Insights into the Hypermethylation of BRCA1 Evinces a Novel Pathway to Breast Tumorigenesis

BackgroundBreast cancer is the most common and dreadful form of cancer globally. Inherited mutations in the breast cancer susceptibility gene, BRCA1, increases the cumulative risk of developing breast cancer by 70‐80%. However, hereditary mutations account for only 5‐10% of the breast cancer burden. At the same time, the rest of this menace is attributed to sporadic cases where patients rarely encounter a BRCA1 mutation. Instead, upto 60% of these patients manifest low BRCA1 expression associated with BRCA1 promoter hypermethylation—moreover, many hereditary cases with a mutated BRCA1 allele exhibit promoter hypermethylation in the second allele. Interestingly, most of these cases also develop into a hyper‐aggressive and drug‐resistant form, Triple Negative Breast Cancer (TNBC), where Estrogen Receptors (ER), Progesterone receptors (PR) and Human epidermal growth factor receptors (Her2) are absent or minimally expressed. But the molecular signaling governing this tumorigenesis has remained elusive with an unresolved question of whether promoter hypermethylation is the master controller of the process of tumorigenesis or is a mere consequence of tumor progression.Objective &amp; MethodologyTo unravel the truth behind this enigma, we induced site‐specific methylations in the CpG island of BRCA1 promoter on wildtype BRCA1 backgrounds using a modified version of CRISPR technology.ResultsInducing site‐specific methylation on the BRCA1 promoter effectuated its downregulation via alteration in the balance between its alternate transcripts ‘β’ and ‘α’. Methylation also attenuated the expression of a long noncoding RNA, NBR2 (Neighbor of BRCA1 gene 2), which is transcribed through the BRCA1 promoter in the reverse direction. It, in turn, activates a feedback loop leading to further downregulation of BRCA1 associated with impaired DNA damage repair and a shift in the balance between apoptosis and autophagy evident under glucose starvation conditions. Further, the β subunit of hCG (human chorionic gonadotrophin) hormone, which is often associated with aggressive forms of BRCA1 mutated breast cancers, was significantly overexpressed. With regards to TNBC progression, methylation also led to the downregulation of the hormone receptors except for ER‐ α, which was found to be overexpressed. This ER‐ α surge helps the cells survive the onslaught following DNA Damage under BRCA1 defective condition.ConclusionFor the first time, this study elucidates the molecular signaling behind breast tumorigenesis involving BRCA1 hypermethylation. Like BRCA1 mutation screening, our findings open up an outstanding opportunity for screening sporadic breast cancers in females using the combination of NBR2, β‐hCG, and BRCA1 hypermethylation as biomarkers from blood. Moreover, as therapeutic intervention, this study also provides scope for the use of BRCA1 hypermethylation reversing drugs like methotrexate, drugs like biguanides that show better action under NBR2 depleted conditions and development of β‐hCG inhibitors for a better prognosis.

A microengineered biomimetic model of the placental barrier to study environmental exposures during pregnancy

Exposure to toxic heavy metals contributes to the development of pregnancy complications that can adversely affect fetal development. Despite increasing evidence suggesting the negative health impact of heavy metals, investigating their adverse effects in the context of human pregnancy remains a major challenge due to the difficulty of human subject research and the limited capacity of animal models to properly represent the anatomy and function of the human reproductive system. Motivated by problem, here we describe a microengineered in vitro model designed to reverse engineer the maternal-fetal interface in the human placenta. This biomimetic system enables advanced capabilities to grow human trophoblast cells and fetal vascular endothelial cells in a physiological spatial arrangement and dynamic culture conditions to engineer the placental barrier in vitro that can emulate regulated material transport between maternal and fetal circulations. Using cadmium (CdCl2 ) as a representative example of heavy metals, we investigate whether and how this environmental chemical affects the human placental barrier and its physiological function as a gatekeeper. Our study also examines the role of membrane transporters in the maternal-to-fetal transfer of cadmium to provide insights into efflux transporter-medicated protection against environmental exposures during pregnancy.We used soft lithography to construct a compartmentalized elastomeric device consisting of two overlapping chambers separated by a thin semipermeable membrane. After sterilization and surface coating with extracellular matrix protein, the device was seeded with BeWo b30 human trophoblast cells and primary human villous endothelial cells on the opposite sides of the membrane. The bi-layer tissue was then cultured under perfusion conditions by using syringe pumps connected to each chamber. To mimic the physiological process of placental development, we treated the trophoblast cell layer in the maternal chamber with forskolin (50 µM) to induce syncytialization.Our microengineered model of the human placental barrier formed a tight barrier (evaluated by expression of E-cadherin and VE-cadherin), secreted the pregnancy hormone human chorionic gonadotropin beta (hCGβ), and supported physiological maternal-to-fetal transport of glucose. Following CdCl2 treatment (0.5-50 µM), we observed concentration-dependent deleterious responses of the maternal and fetal tissues as demonstrated by reduced cell viability and placental hormone secretion, compromised barrier function, and increased secretion of proinflammatory cytokines. The results also indicated upregulated expression of metallothioneins IA and IIA (MT-IA/IIA) metal binding proteins.The placenta-on-a-chip is an innovative and translational in vitro platform for the study of environmental chemical toxicity in the human placenta. With further improvements, this technology may advance our ability to model, interrogate, and predict the potential of xenobiotics to disrupt placentation and maintenance of human pregnancy.

Gestational trophoblastic disease and associated factors among women experiencing first trimester pregnancy loss at a regional referral hospital in central Tanzania: a cross-sectional study.

Gestational trophoblastic diseases (GTDs) may follow any form of pregnancy or a pregnancy loss. Early detection of GTDs is important, as some benign forms of the disease may progress into a chemoresistant and metastatic disease. This study aimed at determining the frequency of GTDs among women experiencing first trimester pregnancy loss and the associated patients' characteristics. This was a cross-sectional study that included 200 conveniently sampled women who experienced first trimester pregnancy loss from January to December 2019 at a Regional Referral Hospital in central Tanzania. The specimen obtained from products of conception were collected, formalin-fixed and paraffin-embedded and submitted for histopathological evaluation, for which haematoxylin and eosin stain was used. Data were analysed using SPSS version 23.0. The χ2 test was used to determine the association between categorical variables. p-Values ˂0.05 were considered statistically significant. Among 200 study participants, the overall frequency of GTDs was 42 (21%). Among those with GTDs, the most common histopathological diagnosis was partial hydatidiform mole (18 [42.9%]), followed by complete hydatidiform mole (17 [40.5%]) and choriocarcinoma (7 [16.5%]). In the studied participants, only increased human chorionic gonadotropin hormone levels were found to be statistically significantly associated with GTDs (p=0.000). Results from this study suggest that routine histopathological evaluation of the products of conception is recommended in order to allow early detection of GTDs, including choriocarcinoma, which usually carries a poor prognosis. The histopathological reporting of choriocarcinoma among first trimester products of conception from Tanzania is novel.

Testicular haemodynamics, plasma testosterone and oestradiol concentrations, and serum nitric oxide levels in the Egyptian buffalo bull after a single administration of human chorionic gonadotropin.

This current study aimed for the first time to evaluate the effect of a single intravenous administration of human chorionic gonadotropin hormone (hCG) on the testicular artery haemodynamics measurements (resistance [RI], and pulsatility indices [PI]), plasma steroids (estradiol-17β and testosterone) and nitric oxide (NO) levels in buffalo bulls. Twelve Egyptian buffalo bulls weighted 450±20kg were randomly divided into the hCG group (n=6) and injected a single dose with Ovogest (EPIFASI; 5,000 IU, iv), whereas the others (n=6) were injected with normal saline and served as controls. Doppler evaluation and blood sampling were performed just before the administration (hour 0) and at 1, 2, 4, 6, 8, 10, 24 and 28hr after administration. Bulls in the control group did not show any alterations in hormonal levels and blood flow parameters (p>.05). In the hCG group, RI was declined (p<.05) in 6hr post-administration (0.31±0.01 versus 0.45±0.01), while PI was declined (p<.05) later in 10hr (0.74±0.01 versus 1.23±0.02). Additionally, testicular blood flow was increased (p<.05) 8hr (42.02±1.02ml/min/100g versus 31.34±0.88ml/min/100g) after administration. Testosterone and NO levels were (p<.05) increased at 4 and 6hr post-administration (3.55±0.03ng/ml versus 2.84±0.01ng/ml, and 55.32±4.25µmol/L versus 32.21±1.55µmol/L), whereas oestradiol levels were elevated (p<.05)in 6hr (31.25±0.08pg/ml) only post-administration then declined. In conclusion, the single intravenous administration of hCG triggered many alterations in the supratesticular artery vascularization and hormonal profile that could affect positively on steroidogenesis and testicular function in buffalo bull.

Continuous Testosterone Therapy with Recombinant FSH & HCG Improves Semen Parameters: A Pilot Study

ABSTRACT Introduction Male hypogonadism is characterized by inadequate production of Testosterone (T) and a deficiency in spermatogenesis. The main treatment of male hypogonadism is T therapy (TTh), which can disrupt normal hypothalamic-pituitary-gonadal axis function and cause oligospermia and/or azoospermia with testicular atrophy. Objective We evaluated the safety and efficacy of continuous intramuscular testosterone administration in combination with low doses of human chorionic gonadotropin (HCG) and recombinant follicle stimulating hormone (FSH) on semen parameters. Methods A total of 10 hypogonadal men on TTh, HCG, and FSH who had achieved pregnancy with their partners or had successful sperm retrievals were retrospectively identified. All of the men were maintained on continuous intramuscular testosterone therapy, HCG at 750 IU subcutaneously twice weekly and recombinant FSH 75 IU every other day. A semen analysis was performed in men who had not had vasectomy prior to or after starting therapy, and serum testosterone, estrogen, FSH, LH, hemoglobin and hematocrit levels, as well as a scrotal ultrasound for testicle size determination, were obtained. Results Of the 10 men with a mean(SD) age of 39.5(6.4) years) who underwent treatment, 7 achieved pregnancy naturally and 3 underwent sperm retrieval due to prior vasectomy, with successful sperm retrieval in all 3. Mean(SD) pretreatment T levels were 745(614) ng/dl, Estradiol 38.7(42.8) pg/ml, LH 0.18(0.24) IU/L, FSH 0.47(0.46) IU/L. Mean(SD) total motile sperm count (TMSC) was 1.43(2.99) million total motile sperm. Mean(SD) pretreatment testicular volume was 11.1(3.3)cc and post-treatment was 11(5.5) cc. Mean(SD) post-treatment T levels were 1185(482) ng/dl, Estradiol 26.0(16.9) pg/ml, LH 0.43(0.23) IU/L, FSH 1.78(0.91) IU/L, with TMSC 102.8(89.4) million. Conclusions Across a cohort of 10 men, TTh in combination with HCG and FSH resulted in resumption or increase in TMSC during treatment, resulting in either natural pregnancy or successful sperm retrieval and supporting the conclusion that TTh does not require cessation in men desiring fertility during TTh. Further prospective studies are required to validate these findings. Disclosure Any of the authors act as a consultant, employee or shareholder of an industry for: Vault Health

Effect of Recombinant Gonadotropin on Testicular Function and Testicular Sperm Extraction in Five Cases of NR0B1 (DAX1) Pathogenic Variants.

Background NR0B1 pathogenic variants can cause congenital adrenal hypoplasia or primary adrenal insufficiency in early childhood usually associated with hypogonadotropic hypogonadism. NR0B1 is necessary for organogenesis of the adrenal cortex and to maintain normal spermatogenesis. In humans, restoration of fertility in patients carrying NR0B1 pathogenic variants is challenging.ObjectiveThe aim of the study was to investigate the clinical, hormonal, histological, spermiological, and molecular genetic characteristics of a cohort of patients with NR0B1 pathogenic variants, monitored for fertility preservation.PatientsWe included five patients, including four teenagers, with NR0B1 pathogenic or likely pathogenic variants. They all had primary adrenal insufficiency and were receiving replacement therapy with glucocorticoids and mineralocorticoids. Patients received recombinant follicle-stimulating hormone and recombinant human chorionic gonadotropin in order to induce spermatogenesis. Combined gonadotropin treatment was initiated between 13 years and 15 years and 6 months for the four teenagers and at 31 years and 2 months for the only adult. Physical and hormonal assessments were performed just before starting gonadotropin treatment. After 12 months of gonadotropin treatment, physical examination and hormonal assessments were repeated, and semen analyses were performed. If no sperm cells were observed in at least 2 semen collections at 3-month interval, testicular biopsy for testicular sperm extraction was proposed.ResultsBilateral testicular volume increased from 8 ml (interquartile range, 6–9) to 12 ml (10–16) after gonadotropin treatment. Inhibin B levels were relatively stable: 110 ng/L (46–139) before and 91 ng/L (20–120) at the end of gonadotropin treatment. Azoospermia was observed in all semen analyses for all cases during gonadotropin treatment. Three patients agreed to testicular biopsy; no mature sperm cells could be retrieved in any.ConclusionWe characterized a cohort of patients with NR0B1 pathogenic or likely pathogenic variants for fertility preservation by recombinant gonadotropin treatment, which began either at puberty or in adulthood. No sperm cells could be retrieved in semen samples or testicular biopsy even after gonadotropin treatment, indicating that gonadotropin treatment, even when started at puberty, is ineffective for restoring fertility.

Success rate of methotrexate in the conservative treatment of tubal ectopic pregnancies.

An ectopic pregnancy (EP) involves the implantation of the gestational sac outside the uterine cavity. In the event of diagnosing an EP, the current medical approach is to avoid surgery and to preserve fertility whenever possible; therefore, methotrexate (MTX)-based therapy has become prominent in recent years. MTX, a drug usually used to treat severe forms of autoimmune diseases and several types of cancer, has proven its utility in the conservative treatment of EPs. The success rate of MTX correlates with lower values of β subunit of human chorionic gonadotropin hormone (β-hCG) serum levels, especially below 2,000 mUI/ml, side effects being insignificant. In the present study, the results obtained concerning the success rate of MTX in the conservative treatment of EPs were obtained at the Department of Obstetrics and Gynecology of the Bucharest University Emergency Hospital from January 2014 to December 2020. The aim of the present study was to highlight the necessity for revising current guidelines for ectopic pregnancy medical treatment in order to manage this pathology optimally and to select carefully the proper treatment, whether medical, surgical or expectant management, so that morbidity is reduced to a minimum.

Long-acting microspheres of Human Chorionic Gonadotropin hormone: In-vitro and in-vivo evaluation

Human Chorionic Gonadotropin (hCG) hormone is used to cause ovulation, treat infertility in women, and increase sperm count in men. Conventional hCG solution formulations require multiple administration of hCG per week and cause patient noncompliance. The long-acting PLGA depot microspheres (MS) approach with hCG can improve patient compliance, increase the efficacy of hCG with a lower total dose and improve quality of life. Therefore, hCG was encapsulated by a modified double emulsion solvent evaporation technique within PLGA MS by high-speed homogenizer and industrially scalable in-line homogenizer, respectively. MS was characterized for particle size, encapsulation efficiency (EE), surface morphology, and in-vitro release. The spherical, dense, non-porous microspheres were obtained with a size of 58.88 ± 0.18 µm. Microspheres showed high EE (77.4% ± 5.9%) with low initial burst release (12.82% ± 2.07%). Circular Dichroism and SDS-PAGE analysis indicated good stability and structural integrity of hCG in the microspheres. Its bioactivity was proven further by a bioassay study in immature Wistar rats. Pharmacokinetic analysis showed that the hCG PLGA MS maintained serum hCG concentration up to 13 days compared to multiple injections of a marketed conventional parenteral injectable formulation of hCG. Thus, it can be ascertained that the hCG PLGA MS may have great potential for clinical use in long-term therapy.