Gonadotropin Production Research Articles

Evaluation of Cardiovascular Disease Risk in Women having Poly Cystic Ovarian Syndrome

PCOS, also known as Stein-Leventhal syndrome, is a common endocrine disease characterized by two of the following three characteristics: Once the associated endocrinological and gynecological diseases have been ruled out, oligo-ovulation or anovulation, ii) clinical and/or biochemical indications of hyperandrogenism, or iii) polycystic ovaries should be considered. Cardiovascular disease (CVD) risk factors are common in women with polycystic ovarian syndrome (PCOS). The Androgen Excess and Polycystic Ovary Syndrome (AE-PCOS) Society established a panel to offer evidence-based evaluations of research examining the PCOS-CVD risk connection and to produce CVD prevention recommendations. The main pathophysiological abnormality in polycystic ovarian syndrome is a source of much debate (PCOS). Chronic anovulation in conjunction with androgen excess, hyperinsulinemia, and changes in gonadotropin production are now widely accepted as symptoms of this disease in women. Polycystic ovarian syndrome (PCOS) is linked to obesity and low-grade inflammation, and it may raise the risk of cardiovascular disease (CVD). This study examines the assessment of cardiovascular disease risk in women with PCOS.

Development of a suitable manufacturing process for production of a bioactive recombinant equine chorionic gonadotropin (reCG) in CHO-K1 cells

Equine chorionic gonadotropin (eCG) is a heterodimeric glycoprotein hormone produced by pregnant mares that has been used to improve reproductive performance in different domestic species. Several strategies to produce the hormone in a recombinant way have been reported; nevertheless, no approach has been able to produce a recombinant eCG (reCG) with significant in vivo bioactivity or in sufficient quantities for commercial purposes. For this reason, the only current product available on the market consists of partially purified preparations from serum of pregnant mares (PMSG). Herein, we describe a highly efficient process based on third-generation lentiviral vectors as delivery method for the production of reCG in suspension CHO-K1 cells, with productivities above 20 IU 106 cell−1.d−1 and 70% purification yields after one purification step. Importantly, reCG demonstrated biological activity in cattle, since around 30 μg of reCG were needed to exert the same biologic effect of 400 IU of PMSG in an ovulation synchronization protocol. The results obtained demonstrate that the developed strategy represents an attractive option for the production of reCG and constitutes an auspicious alternative for the replacement of animals as a source of PMSG.

Reduced FSH and LH action: implications for medically assisted reproduction.

ABSTRACTLuteinizing hormone (LH) and follicle-stimulating hormone (FSH) play complementary roles in follicle development and ovulation via a complex interaction in the hypothalamus, anterior pituitary gland, reproductive organs, and oocytes. Impairment of the production or action of gonadotropins causes relative or absolute LH and FSH deficiency that compromises gametogenesis and gonadal steroid production, thereby reducing fertility. In women, LH and FSH deficiency is a spectrum of conditions with different functional or organic causes that are characterized by low or normal gonadotropin levels and low oestradiol levels. While the causes and effects of reduced LH and FSH production are very well known, the notion of reduced action has received less attention by researchers. Recent evidence shows that molecular characteristics, signalling as well as ageing, and some polymorphisms negatively affect gonadotropin action. These findings have important clinical implications, in particular for medically assisted reproduction in which diminished action determined by the afore-mentioned factors, combined with reduced endogenous gonadotropin production caused by GnRH analogue protocols, may lead to resistance to gonadotropins and, thus, to an unexpected hypo-response to ovarian stimulation. Indeed, the importance of LH and FSH action has been highlighted by the International Committee for Monitoring Assisted Reproduction Technologies (ICMART) in their definition of hypogonadotropic hypogonadism as gonadal failure associated with reduced gametogenesis and gonadal steroid production due to reduced gonadotropin production or action. The aim of this review is to provide an overview of determinants of reduced FSH and LH action that are associated with a reduced response to ovarian stimulation.

Gnrh2 maintains reproduction in fasting zebrafish through dynamic neuronal projection changes and regulation of gonadotropin synthesis, oogenesis, and reproductive behaviors

Restricted food intake, either from lack of food sources or endogenous fasting, during reproductive periods is a widespread phenomenon across the animal kingdom. Considering previous studies show the canonical upstream regulator of reproduction in vertebrates, the hypothalamic Gonadotropin-releasing hormone (Gnrh), is inhibited in some fasting animals, we sought to understand the neuroendocrine control of reproduction in fasted states. Here, we explore the roles of the midbrain neuropeptide, Gnrh2, in inducing reproduction via its pituitary prevalence, gonadotropin synthesis, gametogenesis, and reproductive outputs in the zebrafish model undergoing different feeding regimes. We discovered a fasting-induced four-fold increase in length and abundance of Gnrh2 neuronal projections to the pituitary and in close proximity to gonadotropes, whereas the hypothalamic Gnrh3 neurons are reduced by six-fold in length. Subsequently, we analyzed the functional roles of Gnrh2 by comparing reproductive parameters of a Gnrh2-depleted model, gnrh2−/−, to wild-type zebrafish undergoing different feeding conditions. We found that Gnrh2 depletion in fasted states compromises spawning success, with associated decreases in gonadotropin production, oogenesis, fecundity, and male courting behavior. Gnrh2 neurons do not compensate in other circumstances by which Gnrh3 is depleted, such as in gnrh3−/− zebrafish, implying that Gnrh2 acts to induce reproduction specifically in fasted zebrafish.

Genistein Blunted Detrimental Effects of Polycystic Ovary Syndrome on the Ovarian Tissue of Rats by Improving Follicular Development and Gonadotropin Secretion.

ObjectivePolycystic ovary syndrome (PCOS) is a common cause of female infertility worldwide. It has been shown that genistein, a natural isoflavone, may influence follicular competence via the production of gonadotropins in women with PCOS. The current study aims to evaluate the effects of genistein on the ovarian tissue of rats with PCOS.MethodsThirty female Wistar rats were randomly divided into the following four groups: Control; PCOS (rats received 2 mg/kbW estradiol valerate); Genistein (rats given 1 mg/kg BW of genistein for 14 days); and Genistein + PCOS. All animals were slaughtered under anesthesia and blood samples were collected for biochemical analysis. Follicular morphology was analyzed based on histologic examination.ResultsHistologic examination exhibited enhanced follicular atresia at various stages in the rats with PCOS compared to controls (p<0.001). Induction of PCOS caused significant reduction in gonadotropin levels and steroid hormone levels consistent with insulin resistance (p<0.01). Data showed that 14-day administration of genistein might improve follicular morphology in rats with PCOS (p<0.001). Genistein treatment increased the production of gonadotropins and steroid hormones and alleviated insulin resistance in Rats with PCOS (p<0.001).ConclusionsThis study indicated that genistein treatment exerted a beneficial effect on the ovarian tissue of rats with PCOS by improving follicular growth and hormone balance.

Functional differences in the hypothalamic-pituitary-gonadal axis are associated with alternative reproductive tactics based on an inversion polymorphism

The evolution of social behavior depends on genetic changes, yet, how genomic variation manifests itself in behavioral diversity is still largely unresolved. Chromosomal inversions can play a pivotal role in producing distinct behavioral phenotypes, in particular, when inversion genes are functionally associated with hormone synthesis and signaling. Male ruffs exhibit alternative reproductive tactics (ARTs) with an autosomal inversion determining two alternative morphs with clear behavioral and hormonal differences from the ancestral morph. We investigated hormonal and transcriptomic differences in the pituitary and gonads. Using a GnRH challenge, we found that the ability to synthesize testosterone in inversion carriers is severely constrained, whereas the synthesis of androstenedione, a testosterone precursor, is not. Inversion morphs were able to produce a transient increase in androstenedione following the GnRH injection, supporting the view that pituitary sensitivity to GnRH is comparable to that of the ancestral morph. We then performed gene expression analyses in a second set of untreated birds and found no evidence of alterations to pituitary sensitivity, gonadotropin production or gonad sensitivity to luteinizing hormone or follicle-stimulating hormone across morphs. Inversion morphs also showed reduced progesterone receptor expression in the pituitary. Strikingly, in the gonads, inversion morphs over-expressed STAR, a gene that is located outside of the inversion and responsible for providing the cholesterol substrate required for the synthesis of sex hormones. In conclusion, our results suggest that the gonads determine morph-specific differences in hormonal regulation.

Maternal Obesity Influences Placental Nutrient Transport, Inflammatory Status, and Morphology in Human Term Placenta.

Maternal obesity has a significant impact on placental development. However, this impact on the placenta's structure and function (ie, nutrient transport and hormone and cytokine production) is a controversial subject. We hypothesized that maternal obesity is associated with morphologic, secretory, and nutrient-related changes and elevated levels of inflammation in the placenta. We collected samples of placental tissue from 2 well-defined groups of pregnant women from 2017 to 2019. We compared the 2 groups regarding placental cytokine and hormone secretion, immune cell content, morphology, and placental nutrient transporter expressions. Placenta were collected after caesarean section performed by experienced clinicians at Centre Hospitalier Intercommunal (CHI) of Poissy-Saint-Germain-en-Laye. The main inclusion criteria were an age between 27 and 37 years old, no complications of pregnancy, and a first-trimester body mass index of 18-25 kg/m2 for the nonobese (control) group and 30-40 kg/m2 for the obese group. In contrast to our starting hypothesis, we observed that maternal obesity was associated with (1) lower placental IL-6 expression and macrophage/leukocyte infiltration, (2) lower placental expression of GLUT1 and SNAT1-2, (3) a lower placental vessel density, and (4) lower levels of placental leptin and human chorionic gonadotropin production. These results suggest that the placenta is a plastic organ and could optimize fetal growth. A better understanding of placental adaptation is required because these changes may partly determine the fetal outcome in cases of maternal obesity.

Differences in invitro responses of the hypothalamo–pituitary–gonadal hormonal axis between low- and high-egg-producing turkey hens

Low-egg-producing hens (LEPH) ovulate less frequently than high-egg-producing hens (HEPH) and exhibit differences in mRNA levels for components of the hypothalamo–pituitary–gonadal (HPG) axis, suggesting differential responsiveness to trophic stimulation. Ovulation frequency is governed by the production of the pituitary gonadotropins and feedback of the ovarian follicle steroid hormones, which are regulated by HPG axis stimulation and inhibition at the hypothalamic level. The pituitary and follicle cells from LEPH and HEPH were subjected to in vitro hormonal treatments to stimulate or inhibit the HPG axis, followed by expression analysis of mRNA levels for HPG axis genes and radioimmunoassays for steroid hormone production. Statistical analysis was performed using the mixed models procedure of SAS. The pituitary cells from HEPH showed upregulation of genes associated with ovulation stimulation, whereas cells from LEPH showed upregulation of genes associated with inhibition of ovulation. High-egg-producing hens’ follicle cells displayed a higher sensitivity and responsiveness to gonadotropin treatment. Level of egg production impacted ovulation-related gene expression in the pituitary cells as well as steroid hormone production in the follicle cells, with HEPH displaying a greater positive response to stimulation. These findings indicate that differences in egg production among turkey hens likely involve differential responsiveness of the cells within the HPG axis.

Interference of gadolinium dechelated from MR contrast agents by calcium signaling in neuronal cells of GnRH.

Contrast agents (CAs) used in magnetic resonance imaging (MRI) are produced by chelating the metal gadolinium (Gd) with organic ligand molecules to form stable complexes. But, Gd3+ may dissociate from the CAs and subsequently might become toxic to its environment. Besides toxicity, it might inhibit calcium channels on cell membranes and this action could be detrimental to the cells governing biological development. The aim of this study was to investigate the interference of Gd3+ dechelated from the CAs bycalcium signaling in the neuronal cells of gonadotropin-releasing hormone (GnRH), regulating puberty, and sexual development. The study used the mouse GT1-7 cell line as a model system, and Fura-2 based calcium imaging for detecting the interruption of intracellular calcium transport by the extracellular presence of Gd3+ as released from the CAs; gadodiamide and gadoterate meglumine, when the cells were stimulated invitro culture by exposure to melatonin.The CA gadoterate meglumine interfered minimally with the calcium signaling, and thus its use is preferable in standard MRI exams. The release of Gd3+ from gadodiamide was significant and becomes of great concern as it may impact the neurophysiology of the neuronal cells in general,and gonadotropin production in particular, even in normal patients without nephrogenic systemic fibrosis. The toxicity induced by the influx of dechelated Gd3+ in the neurons of GnRH would have significant implications for puberty and reproductive functions.

SUN-513 Severe Hyperthyroidism in a Complete Molar Pregnancy

Hydatidiform mole (HM), a type of gestational trophoblastic disease (GTD), is a rare cause of clinical hyperthyroidism. The development of hyperthyroidism requires an elevation of HCG >100,000 mlU/mL for several weeks. Complete mole has a marked HCG elevation compared to partial mole thus presents with a higher incidence of thyrotoxicosis. Surgical uterine evacuation is the treatment of choice for HM. However, untreated hyperthyroidism can pose a risk for the development of thyroid storm and high-output cardiac failure in the perioperative period. To our knowledge, there are no specific guidelines for management at this time. We present a case of hyperthyroidism secondary to complete molar pregnancy successfully treated with propylthiouracil (PTU), potassium iodide (SSKI), and atenolol in the preoperative period.A 42-year-old female with history of migraines presented to her gynecologist with a 3-week history of lower abdominal cramping, vomiting, loss of appetite, and abnormal vaginal bleeding. She also endorsed a 6-pound weight loss, intermittent tachycardia, exertional dyspnea, and increased anxiety. Pregnancy test was positive, and ultrasound was concerning for GTD. Laboratory work up was significant for HCG 797,747 mIU/mL (< 5mlU/mL), TSH <0.005 mIU/mL (0.4-4.0 mlU/mL), Free T4 3.09 ng/dL (0.9-1.9 ng/dL), and Free T3 11.48 pg/dL (1.76-3.78 pg/dL). The patient was admitted to the hospital and started on PTU 100 mg Q6H, SSKI 200 mg TID following the first dose of PTU, and atenolol 25 mg daily. She underwent an uncomplicated D & C the next day. On post-op day 1, HCG decreased to 195,338 mIU/mL and Free T4 to 2.39 ng/dL. The patient was discharged on the aforementioned doses of PTU and atenolol. One-week follow-up labs showed HCG 8,917 mIU/mL and Free T4 1.22 ng/dL. Surgical pathology confirmed a complete hydatidiform mole. PTU was decreased to 50 mg TID. On post-op day 14, HCG had risen to 15,395 mIU/mL with onset of nausea and vomiting. Repeat Free T4 remained within reference range. Patient was taken back to surgery for a laparoscopic total hysterectomy with bilateral salpingectomy. Pathology confirmed an invasive hydatidiform mole. Two-week follow-up lab work showed HCG 155 mIU/mL, TSH 1.5 mIU/mL, and Free T4 1.19 ng/dL. PTU and atenolol were then discontinued.The development of hyperthyroidism in molar pregnancy is largely influenced by the level of HCG and usually resolves with treatment of GTD (1). However, it’s crucial to control thyrotoxicosis to avoid perioperative complications. This case also highlights the importance of monitoring HCG levels following a complete molar pregnancy due to an increased risk for invasive neoplasm.1. Walkington, L et al. “Hyperthyroidism and human chorionic gonadotrophin production in gestational trophoblastic disease.” British journal of cancer vol. 104,11 (2011): 1665-9. doi:10.1038/bjc.2011.139

SUN-036 Evaluation of Gender Experience Among Individuals with Isolated GnRH Deficiency Compared to Controls

Background: Sex hormones play a role in gender identity development. For example, 46,XY individuals with complete androgen insensitivity typically have a female gender identity. Isolated GnRH deficiency (IGD) leads to hypogonadism due to decreased GnRH-induced gonadotropin production. It is unknown if decreased sex hormone exposure leads to differences in gender identity among individuals with IGD compared with the general population. Our objective was to determine if the gender identity in subjects with IGD differs from controls.Methods: We distributed a validated questionnaire; the Gender Identity/Gender Dysphoria Questionnaire for Adults and Adolescents (GIDYQ-AA) (1), to IGD participants who previously enrolled in a phenotyping study. Subjects also provided their age, sex assigned at birth (SAAB), gender and information about their condition and treatment. Group survey scores are expressed as mean ± SD. IGD subject scores were compared with control data (Student’s t-test) obtained from a validation study for the GIDYQ-AA (2).Results: Out of 79 subjects who were contacted, 8 males (M) assigned at birth and 7 females (F) assigned at birth chose to participate and one person actively declined. Average age was 30 y for F and 28 y for M. At the time of the study, all subjects were on hormone supplementation except for one F. A score of 5 indicates a gender identity congruent with SAAB. Among F, mean scaled IGD score was 4.73 ± 0.29 vs. controls (4.8 ± 0.28, n=57; not significant (ns)), and for M it was 4.62 ± 0.52 vs. controls (4.82 ± 0.24, n=37; ns). One female identified as “non-gendered”, one male identified as “intersex/DSD” (14% of respondents), one female did not respond and gender was congruent with SAAB in the remainder. The lowest mean score for an individual question for the IGD F group was in response to whether they felt satisfied being a woman and for the IGD M group, it was in response to whether they felt they were a real man.Conclusions: Men and women with IGD did not show a significant difference in their gender identity compared with controls, and gender was found to be congruent with SAAB for the majority. However, the individual question responses and the self-described gender in this small cohort may suggest that there are differences in how some individuals with IGD experience their gender development. We speculate that this may be related to how they perceive the differences in physical development that they experienced related to their condition. Larger studies in participants with IGD and other disorders that alter sex hormone production/effect are necessary to further understand the relationship between decreased sex hormone exposure during critical developmental periods and gender identity development.

SAT-292 Musashi: A Novel Regulator of the Gonadotrope Transcriptome

Sufficient nutrition is critical for reproduction. We have previously shown that leptin, a circulating indicator of fat stores, signals to pituitary gonadotropes to maintain gonadotropin releasing hormone receptor (GnRHR) protein levels in female mice. We hypothesized that this process is post-transcriptional, happening primarily through regulation of the RNA-binding protein Musashi (MSI). We showed that MSI binds to Gnrhr and inhibits translation, and a gonadotrope-specific deletion of Msi1 and Msi2 (Gon-Msi1/2-null) leads to increased GnRHR protein levels. This culminates in dysregulated luteinizing hormone (LH) and follicle-stimulating hormone (FSH). We have recently identified other gonadotrope and pituitary targets of MSI. We therefore suspected that MSI plays a role in both the maturation of gonadotropes and the normal cyclic regulation of gonadotropes. We hypothesized that the deletion of MSI would lead to downstream effects on (1) the composition of the gonadotrope population and (2) the molecular landscape of these cells. Using our adult, diestrous Gon-Msi1/2-null females, we performed single-cell RNA-sequencing on methanol-fixed dispersed pituitary cells. Libraries were made from two control pools and two mutant pools (n=3 pituitaries/pool) using 10x Genomics v3.1 Single-Cell Gene Expression technology and initially sequenced on an Illumina Next-seq mid-output flow-cell, yielding 5,000 reads/cell. Subsequent high-output sequencing obtained 25,000 reads/cell. We recovered single-cell mRNA transcript information from 18,206 control pituitary cells and 16,255 Gon-Msi1/2-null cells. Our analyses revealed that the Gon-Msi1/2-null pools had a higher % of cells expressing Fshb, as well as an expected significant drop in Msi2-expressing gonadotropes and no change in Lhb-expressing cells. We have recently identified Fshb as an MSI target in silico, and qRT-PCR of female pituitary lysate immunoprecipitated with anti-MSI1 shows a 7-fold enrichment in Fshb mRNA. We identified differentially expressed genes comparing the control and Gon-Msi1/2-null gonadotrope clusters. Using Gene Ontology analyses, the Gon-Msi1/2-null gonadotrope cluster appears to have aberrant expression of mRNAs involved in protein folding and cellular responses to nutrients. Our high-output sequencing has allowed us to achieve 25,000 reads/cell and will provide greater resolution of the role of Musashi in control of gonadotrope function. Taken together, our data indicate that Musashi influences the molecular landscape and subsequent physiology of the female gonadotrope. We have identified potential gonadotrope-specific MSI targets, including pathways that may underlie the dysregulated gonadotropin production and secretion seen in our Gon-Msi1/2-null females. Future studies will compare pubertal and adult females, as well as females from different estrous cycle stages.

Biomarkers of male hypogonadism in childhood and adolescence.

The objective of this review was to characterize the use of biomarkers of male hypogonadism in childhood and adolescence. The hypothalamic-pituitary-gonadal (HPG) axis is active during fetal life and over the first months of postnatal life. The pituitary gland secretes follicle stimulating hormone (FSH) and luteinizing hormone (LH), whereas the testes induce Leydig cells to produce testosterone and insulin-like factor 3 (INSL), and drive Sertoli cells to secrete anti-Müllerian hormone (AMH) and inhibin B. During childhood, serum levels of gonadotropins, testosterone and insulin-like 3 (INSL3) decline to undetectable levels, whereas levels of AMH and inhibin B remain high. During puberty, the production of gonadotropins, testosterone, and INSL3 is reactivated, inhibin B increases, and AMH decreases as a sign of Sertoli cell maturation. Based on our knowledge of the developmental physiology of the HPG axis, these biomarkers can be used in clinical practice to interpret the physiopathology of hypogonadism. Additionally, these markers can have diagnostic value in different forms of hypogonadism that may appear during childhood and adolescence.

Identification and expression of gonadotrophin hormones in gouramy (Osphronemous gouramy, Lacepède, 1801) under photoperiod manipulations

Abstract. Prayogo NA, Siregar AS, Sukardi P, Bessho Y. 2020. Identification and expression of gonadotrophin hormones in gouramy (Osphronemoous gouramy, Lacepède, 1801) under photoperiod manipulations. Biodiversitas 21: 1365-1372. The environment influences fish reproduction. One of the influential environmental factors is photoperiod that regulates the production of hormones for gonad growth and development, gametogenesis, then the reproductive cycle. This research was conducted to determine the effect of photoperiod on the reproductive performance of gouramy with different photoperiod treatments. The design of this study was four photoperiod treatments consisting of 14L: 10D, 16L: 8D and 18L: 6D and controls. Fish samples were divided into 4 aquariums consisting of nine fish and repetition was conducted on each. Fish were kept under photoperiod treatment for 16 weeks. The observed variable was pituitary activity. Pituitary activity was evaluated by measuring the expression of FSH and LH genes using Real Time PCR. The results showed that gouramy had FSH and LH genes with 483 bp and 506 bp. FSH and LH gene expression increased with long photoperiod exposure (P &lt;0.05). These findings indicate that photoperiods regulate the reproduction of gouramy through the HPG axis and control the production of the hormone Gonadotropin.

Dioxin male rat reproductive toxicity mode of action and relative potency of 2,3,7,8-tetrachlorodibenzo-p-dioxin and 2,3,7,8-tetrachlorodibenzofuran characterized by fetal pituitary and testis transcriptome profiling

Fetal rat exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) reduces epididymal sperm number involving altered pituitary-testicular hormonal signaling as the proposed mode-of-action (MOA). To evaluate this MOA and compare TCDD to 2,3,7,8-tetrachlorodibenzofuran (TCDF), an in utero rat exposure and study was conducted. Endpoints included congener tissue levels and transcriptomes of maternal liver and fetal liver, testis, and pituitary. Decreased gonadotropin subunit mRNAs levels (Lhb and Fshb) and enriched signaling pathways including GNRH Signaling and Calcium Signaling were observed in fetal pituitary after TCDD (but not TCDF) exposure. TCDD (but not TCDF) decreased fetal testis cholesterologenic and steroidogenic pathway genes. TCDD tissue concentrations in dam liver, dam adipose, and whole fetus were approximately 3- to 6-fold higher than TCDF. These results support a MOA for dioxin-induced rat male reproductive toxicity involving key events in both the fetal pituitary (e.g., reduced gonadotropin production) and fetal testis (e.g., reduced Leydig cell cholesterologenesis and steroidogenesis).

Церебральная микроангиопатия (болезнь мелких сосудов) и возрастной гипогонадизм у мужчин

The review discusses the contribution of age-related male hypogonadism to the formation of cerebral microangiopathy (disease of small brain vessels) (CMA/SVD). The current explanation of the nature of CMA by an ischemic-hypoxic process, which is based on hypo- or hyperperfusion of the white matter of brain due to a complex of various causes, does not explain the etiopathogenesis of CMA. The article considers a violation of the neuroglyovascular connection, represented by a vessel, astrocytic glia, and a neuron, as the cause of SVD development. The main role in the CMA formation is assigned to endothelial dysfunction, all variants of which (vasomotor, thrombophilic, adhesive, angiogenic) can be traced in SVD. The role of disorders of the main functions of arterial vessels — conducting and damping, as well as liquorodynamics disorders in the CMA pathogenesis is discussed. The data indicating the involvement of testosterone and its derivatives in the formation of cerebral vascular pathology are presented. It is emphasized that this participation can be of one kind in men with early physiological uncomplicated hypogonadism, when the hypothalamic-pituitary system is involved in the process, and of another kind in complicated or late climax. At that, an increase in the production of gonadotropin — releasing hormone (GnRH), the content of FSH and LH in peripheral blood is considered as a reaction to an age-related decrease in androgen function, and the vegetative dysfunction, metabolic and other disorders inherent in this period of life – as a result of changes in the functioning of the entire hypothalamus-pituitary-testicular axis. In complicated age-related hypogonadism, the resulting endocrine or somatic pathology is included in the process of small vessel disease formation. In late hypogonadism, the androgen function fades, and the hypothalamic-pituitary system begins to function in a different mode. Often, the clinic and mechanism of CMA formation are determined by diseases inherent in old age. The differences in the pathogenetic mechanisms of CMA development at various periods of IgG determine the need for a differentiated approach to the preventive means and methods of CMA therapy in men of the involutional period. The presented information may be useful for better understanding of CMA development in individuals with age-related hypogonadism.

СРАВНИТЕЛЬНЫЙ АНАЛИЗ СТОМАТОЛОГИЧЕСКОГО СТАТУСА БЕРЕМЕННЫХ ЖЕНЩИН

Currently, oral diseases require an interdisciplinary approach to solving the problem. In most cases, patients go to a doctor during an acute stage or exacerbation of the disease. The high prevalence and increase of inflammatory oral diseases among pregnant women is one of the most significant social problems. Inflammatory processes in the oral cavity can be a provoking factor for the inflammatory reaction of the whole body and subsequently affect the development of the fetus. During pregnancy, a number of changes occur in the oral cavity in the woman's body: the microbiological picture changes, the pathogenicity of the microflora increases, the hormonal background is rebuilt: the production of progesterone, chorionic gonadotropin and other hormones increases. All these factors can lead to changes in the dentate system: deterioration of oral hygiene, increased salivation, halitosis, the appearance of gum bleeding and a change in the pH of saliva. We presented data on the examination of pregnant women in the Tambov Region. We assessed dental status of pregnant women based on dental indices: CPITN, PMA and H.R. Muhleman’s gum bleeding index, estimates of changes in saliva pH of pregnant women.

Serum Levels of Reproductive Hormones and Their Relationship with Age in Men with Benign Prostatic Hyperplasia in Benin City, Edo State

Background: Benign Prostatic Hyperplasia (BPH) is a common condition which occurs as men get older. Aim: We aimed at investigating the reproductive hormone profile of men with BPH and the relationship of these hormones with age. Methods: Twenty-nine men of age > 40 years were screened for prostate symptoms. The prostate examination included digital rectal examination and measurement of serum prostate-specific antigen levels. Demographic and clinical characteristics of subjects were obtained using a structured questionnaire. The serum levels of Testosterone (Te), Luteinizing hormone (LH), Follicle stimulating hormone (FSH) and Prolactin (PRL) were measured using ELISA method. Results: Our study indicated that age and levels of prostate specific antigen (PSA), LH and FSH were significantly (p < 0.05) higher in men with BPH compared with healthy controls. In contrast, the control group indicated significantly (p < 0.05) higher Te level compared with the BPH group. In patients with BPH, age was significantly (p < 0.05) correlated with the hypothalamic-pituitary-testicular axis hormones (LH, FSH and PRL), but not with the sex hormone, and testosterone. Majority (62.1%) of the patients with BPH indicated they had poor libido, while none of them indicated regular sexual activity with their spouses. Conclusion: This study suggests that the rise in serum levels of the hypothalamic-pituitary-testicular axis hormones in BPH patients as demonstrated by the present findings may be age-related since testicular dysfunction in aging men is associated with the decrease in androgen production which may stimulate an increased production of gonadotropins by negative feedback.

90 Establishment and characterization of Day 30 equine chorionic girdle and allantochorion cell lines

Establishing cell lines is a good model for experimental applications to study molecular mechanisms and cell-specific gene expression. Equids have a diffuse epitheliochorial placenta, where the invasive trophoblast is represented by the chorionic girdle (CG) and the noninvasive trophoblast by the allantochorion (AC). Embryonic CG cells are unique to horses and have a crucial role in equine chorionic gonadotropin (eCG) production and maintenance of pregnancy during the first trimester. This study had three objectives: (1) establishing a stable cell line from Day 30 CG cells and AC using lentivirus encoding hTERT; (2) characterisation of Day 30 CG cells and AC cell morphology and expression of eCG α (eCGA) and β (eCGB) subunits, major histocompatibility complex class II (MHCII), and Kisspeptin receptor (KISS1R) in CG and AC cells; (3) investigating eCG protein production invitro from Day 30 CG and AC cells. Three mares (n=3) were used to collect Day 30 conceptuses by non-surgical uterine lavage on Day 30 of pregnancy. All 3 conceptuses were dissected for CG and AC cells then cultured invitro to confluency in cell culture plates. Second-generation lentiviral particles were generated using a three-vector system including transfer vector pLV-hTERT-IRES-hygro, and human telomerase reverse transcriptase (hTERT) lentivirus was utilised to establish stable hygromycin-resistant equine embryonic cell lines. Reverse-transcription PCR (RT-PCR) was used to study gene expression in cells and radioimmunoassay was used to investigate protein presence in the media. We established a hygromycin-resistant Day 30 CG and AC cell lines that express eCGA, eCGB, and hTERT and confirmed using RT-PCR yielding the predicted bands. The cell lines were maintained for 16 passages (7±2 days/passage), 10 of which were cultured after the lentiviral infection steps. Also, we characterised CG cells as fast-growing, large, binucleated, and epithelioid, and AC cells as rapid-growing showing smaller, squamous, mononucleate, epithelioid, and elongated fibroblastic cells. The RT-PCR results showed eCGA and eCGB subunits are expressed by both Day 30 CG and AC cells, but MHCII and KISS1R genes were not expressed in either of cells. Moreover, radioimmunoassay results showed that Day 30 CG cells did produce eCG protein (35.42ngmL−1) invitro earlier than what previous literature has shown. However, Day 30 AC cells did not produce eCG protein (0.042ngmL−1) invitro, and both CG and AC cell lines stopped secreting eCG in the media after the lentiviral infection. To conclude, establishing stable and hygromycin-resistant cell lines from Day 30 equine CG and AC cells using lentivirus encoding pLV-hTERT-IRES-hygro is attainable. Also, equine chorionic gonadotropin eCG protein is produced invitro as early as Day 30 from CG cells.

Horse Welfare During Equine Chorionic Gonadotropin (eCG) Production.

Simple SummaryEquine chorionic gonadotropin or eCG is an important hormone produced by the placenta of pregnant mares and extracted from the blood of these same mares. This hormone is commonly used to enhance reproduction of pigs, dairy cows, sheep, beef cows, and goats. There are currently no alternative sources of this hormone. Horse welfare problems may arise if too much blood is collected at one time or during repeated collections or if the mares are not managed well. In some countries, mares are aborted several months into the pregnancy to improve efficiency since this permits them to become pregnant a second time in one year. We discuss approaches to protect the welfare of pregnant mares kept for eCG production.Collection of blood from pregnant mares for extraction of equine chorionic gonadotropin (eCG) is a critical but relatively unknown and poorly regulated practice in the countries in which it occurs. Equine chorionic gonadotropin is a hormone that is widely used to enhance reproductive performance and management of dairy and beef cattle, sheep, goats, and pigs kept under intensive housing systems. eCG is extracted from the blood of brood mares between days 40–120 of gestation. Although alternatives have been sought, there is currently no efficacious replacement, natural or synthetic, for eCG. Recently, several animal welfare organizations have voiced concerns over the condition and treatment of pregnant mares kept for eCG production in some countries. Animal welfare issues may arise if mares are bled too frequently or if too much blood is collected at any time. In addition, these mares tend to be managed extensively on pastures with minimal veterinary oversight and they may be poorly desensitized and habituated to handling and other practices. This can lead to serious injuries and even death when mares are brought in for bleeding. This paper reviews the process of blood collection for eCG extraction and provides recommendations for ensuring mare welfare.