Abstract Steroids and gonadotrophins are essential for the regulation of late stages of preantral development and antral follicular development. Although the luteinizing hormone receptor (LHCGR) has been detected in the preantral follicles of rats, rabbits, and pigs, its expression, in bovine fetal ovary, has not been demonstrated. Based on this, we aimed to investigate the expression of the LHCGR and LHCGR mRNA binding protein (LRBP), as well as, to quantify bta-miR-222 (a regulatory microRNA of the LHCGR gene) during the development of bovine fetal ovary. In summary, LHCGR expression was observed in the preantral follicle in bovine fetal ovary, from oogonias to primordial, primary and secondary stages, and the mRNA abundance was lower on day 150 than day 60. However, the mRNA abundance of LRBP followed the opposite pattern. Similar to LRBP, the abundance of bta-miR-222 was higher on day 150 than day 60 or 90 of gestation. The LHCGR protein was detected in oogonia, primordial, primary, and secondary follicles. Moreover, both oocytes and granulosa cells showed positive immunostaining for LHCGR. In conclusion, we suggest the involvement of LHCGR/LRBP/bta-mir222 with mechanisms related to the development of preantral follicles in cattle. (LHCGR rats rabbits pigs ovary demonstrated this , (LRBP) btamiR222 btamiR bta miR 222 bta-miR-22 a gene summary primordial 15 However pattern 6 9 gestation oogonia Moreover conclusion LHCGR/LRBP/btamir222 LHCGRLRBPbtamir222 LHCGRLRBPbtamir LHCGR/LRBP/bta mir222 mir LHCGR/LRBP/bta-mir22 cattle (LRBP btamiR22 22 bta-miR-2 1 btamir222 btamir LHCGR/LRBP/btamir22 LHCGRLRBPbtamir22 LHCGRLRBPbta mir22 LHCGR/LRBP/bta-mir2 btamiR2 2 bta-miR- btamir22 LHCGR/LRBP/btamir2 LHCGRLRBPbtamir2 mir2 LHCGR/LRBP/bta-mir bta-miR btamir2 LHCGR/LRBP/btamir

Abstract This review aims to discuss the main factors involved in the development of early antral follicles until gonadotropin dependence. This follicular phase is characterized by intense proliferation of granulosa cells, formation of a fluid-filled cavity, morphological differentiation of cumulus cells, mural granulosa cells and recruitment of theca cells. The interaction between oocyte, granulosa and theca cells is crucial for follicular growth and hormone production. Growth factors produced by the oocyte, such as growth and differentiation factor-9 (GDF-9) and bone morphogenetic protein-15 (BMP-15), regulate granulosa cell proliferation and differentiation and antral cavity development, as well as stimulate the production of follicle-stimulating hormone (FSH) receptors in granulosa cells. In response to FSH, granulosa cells secrete C-type natriuretic peptide (CNP), which acts through its receptor to increase cyclic guanosine monophosphate (cGMP) production and consequently follicular development. Granulosa cells also produce insulin-like growth factor-1 (IGF-1) and increase aromatase enzyme activity, which results in greater sensitivity to gonadotropins and follicular steroidogenesis. The absence of IGF-1 signaling causes cessation of follicular growth at the early antral stage. Many other local factors are involved in the regulation of follicular development. Therefore, this review brings relevant data for a better understanding of the mechanisms involved in the control of early antral follicle growth, emphasizing the role of endocrine and paracrine factors, the oocyte-granulosa cell interaction and the processes of follicular atresia. The challenges for the establishment of efficient culture systems for in vitro growth of early antral follicles are also discussed. dependence fluidfilled fluid filled oocyte factor9 factor 9 factor- GDF9 GDF (GDF-9 protein15 protein 15 protein-1 BMP15, BMP15 BMP , (BMP-15) folliclestimulating stimulating FSH (FSH Ctype C type CNP, CNP (CNP) cGMP (cGMP insulinlike insulin like factor1 1 IGF1 IGF (IGF-1 activity steroidogenesis IGF- stage Therefore oocytegranulosa atresia discussed (GDF- protein1 protein- BMP1 (BMP-15 (CNP (IGF- (GDF (BMP-1 (IGF (BMP- (BMP

Resumo Esta revisão tem como objetivo discutir os principais fatores envolvidos no desenvolvimento de folículos antrais iniciais até a dependência de gonadotrofinas. Essa fase folicular é caracterizada por intensa proliferação de células da granulosa, formação de uma cavidade preenchida por líquido, diferenciação morfológica das células do cumulus, células da granulosa murais e recrutamento de células da teca. A interação entre oócito, células da granulosa e da teca é determinante para o crescimento folicular e produção hormonal. Fatores de crescimento produzidos pelo oócito, fator de crescimento e diferenciação-9 (GDF-9) e proteína morfogenética óssea-15 (BMP-15), regulam a proliferação e diferenciação de células da granulosa, e o desenvolvimento da cavidade antral, bem como estimulam a produção de receptores do hormônio folículo estimulante (FSH) nas células da granulosa. Em resposta ao FSH, as células da granulosa secretam o peptídeo natriurético tipo C (CNP), que atua através de seu receptor para aumentar a produção de monofosfato de guanosina cíclico (GMPc) e consequentemente o desenvolvimento folicular. As células da granulosa também produzem o fator de crescimento semelhante à insulina 1 (IGF-1) e aumentam a atividade da enzima aromatase, o que resulta em maior sensibilidade às gonadotrofinas e esteroidogênese folicular. A ausência de sinalização do IGF-1 causa cessação do crescimento folicular no início do estágio antral. Muitos outros fatores locais estão envolvidos na regulação do desenvolvimento folicular. Por tanto essa revisão traz dados relevantes para uma melhor compreensão dos mecanismos envolvidos no controle do crescimento de folículos antrais iniciais, enfatizando o papel dos fatores endócrinos e parácrinos, a interação oócito-células da granulosa e os processos de atresia folicular. Os desafios para o estabelecimento de sistemas de cultivo eficientes para o crescimento in vitro de folículos antrais iniciais também são discutidos. líquido cumulus oócito hormonal diferenciação9 9 diferenciação- GDF9 GDF (GDF-9 óssea15 óssea 15 óssea-1 BMP15, BMP15 BMP , (BMP-15) antral FSH (FSH CNP, CNP (CNP) GMPc (GMPc IGF1 IGF (IGF-1 aromatase IGF- parácrinos oócitocélulas discutidos (GDF- óssea1 óssea- BMP1 (BMP-15 (CNP (IGF- (GDF (BMP-1 (IGF (BMP- (BMP

SUMMARY OBJECTIVE: The objective of this study was to investigate the allele frequencies of polymorphisms in genes CYP11A1 rs4886595 and CYP11A1 rs4887139 that are responsible for the steroidogenesis mechanism in polycystic ovary syndrome patients and control females. METHODS: Samples were obtained from the Department of Obstetrics and Gynecology in the Near East University Hospital from September 2019 to December 2019. Only the nonobese patients between the ages of 18–40 years were included in this study following informed consent. Obese patients and patients more than 40 years of age were excluded from the study. Nonobese women and normal ovulation were included in the control group. DNA was isolated from blood samples. Real-time polymerase chain reaction (PCR) was used to analyze single nucleotide polymorphisms (SNPs) in various genes linked to polycystic ovary syndrome. The studies were carried out using the samples obtained from 120 women, of whom 55 were nonobese and had normal ovulation, and 65 were polycystic ovary syndrome patients. The allelic frequencies of SNPs in genes linked to polycystic ovary syndrome were calculated using real-time PCR outcomes. RESULTS: The variation of the CYP11A1 rs4887139 G>A did not show any significance, while the variation of CYP11A1 rs4886595 C>A showed significant differences between the patient and the control groups (p=0.01), respectively. CONCLUSION: Future research ought to focus on elucidating the susceptible causes of polycystic ovary syndrome with a wide range of SNPs and more sample size. The genome-wide association studies in polycystic ovary syndrome patients of different origin will be important to identify candidate genes as well as proteins that are implied in polycystic ovary syndrome risk. OBJECTIVE CYPA CYP A CYP11A rs rs488659 rs488713 females METHODS 201 1840 18 18–4 consent 4 group Realtime Real time (PCR (SNPs 12 5 6 realtime real outcomes RESULTS GA G significance CA C p=0.01, p001 p p=0.01 , 0 01 (p=0.01) respectively CONCLUSION size genomewide genome risk rs48865 rs48871 20 184 1 18– p00 p=0.0 (p=0.01 rs4886 rs4887 2 p0 p=0. (p=0.0 rs488 p=0 (p=0. rs48 p= (p=0 rs4 (p= (p

ABSTRACT. Litter size is one of the crucial factors in livestock production and is of high economic value, which is affected by ovulation rate, hormones, and growth factors. Growth factors play a multifaceted role in reproductive physiology. This review aims to investigate the association of bone morphogenetic protein 15 (BMP15) and growth differentiation factor 9 (GDF9) with litter size in livestock. The transforming growth factor β (TGF- β) superfamily includes more than 34 members; GDF9 and BMP15 are among the most significant factors for regulating fertility and litter size in most livestock species. Ovarian follicles release BMP15 and GDF9 that are involved in the maturation of primary follicles into the basal form, proliferation of granulosa and theca cells, steroidogenesis, ovulation, and formation of the corpus luteum. Besides, these factors are highly expressed in oocytes and are necessary for female fertility and multiple ovulation in several livestock species. Animals with two inactive copies of these factors are sterile, while those with one inactive copy are fertile. Thus, the present review provides valuable information on the association of BMP15 and GDF9 with litter size in livestock that can be used as biological markers of multiple ovulation or for improving fertility in livestock. ABSTRACT value rate hormones physiology 1 BMP (BMP15 GDF (GDF9 TGF (TGF 3 members BMP1 species form cells steroidogenesis luteum Besides sterile fertile Thus (BMP1 (GDF (BMP

This study aimed to evaluate in vitro the possible mechanisms underlying the estrogenic potential of benzalkonium chloride (BAC) as a disinfectant emerging contaminant. Effects of BAC at the environmentally-relevant concentrations on estrogen synthesis and estrogen receptor (ER) signaling were assessed using the H295R steroidogenesis assay and the MCF-7 proliferation assay, respectively. Results showed that exposure to BAC at concentrations of 1.0-1.5 mg/L for 48 h significantly increased estradiol production of H295R cells in a concentration-dependent manner. Transcription of steroidogenic genes 3β‐HSD2, 17β‐HSD1, 17β‐HSD4, and CYP19A were significantly enhanced by BAC. In ER-positive MCF-7 cells, exposure to 0.5-1.5 mg/L BAC for 48 h significantly promoted cell proliferation and increased the expressions of ERα and G-protein coupled estrogen receptor 1. Flow cytometry analysis showed that 0.5-1.5 mg/L BAC significantly decreased the percentage of cells in G0/G1 phase, increased the percentage in S phase, and BAC at concentrations of 1.0 and 1.5 mg/L increased the G2/M phase cells. Findings of the study suggested that BAC at environmentally-relevant concentrations might act as a xenoestrogen through its inhibitory effect on steroidogenesis and ER-mediated mechanism. (BAC contaminant environmentallyrelevant environmentally relevant ER (ER HR H R MCF7 MCF 7 MCF- respectively 1.01.5 1015 1 0 5 1.0-1. mgL mg L 4 concentrationdependent concentration dependent manner 3βHSD2 βHSD 3β HSD2 β HSD 3β‐HSD2 17βHSD1 17β HSD1 17β‐HSD1 17βHSD4 HSD4 17β‐HSD4 CYPA CYP A ERpositive positive 0.51.5 0515 0.5 0.5-1. Gprotein G protein G0G1 GG G0 G1 G0/G 10 15 G2M GM G2 M ERmediated mediated mechanism 01 1.01. 101 1.0-1 3βHSD 3β‐HSD 17βHSD 17β‐HSD 51 0.51. 051 05 0. 0.5-1 G0G 1.01 1.0- 0.51 0.5-

Abstract Letrozole comprises a non-steroid aromatase inhibitor that has been applied as a preventive for many uses, such as breast cancer prevention, treatment of hormonal dysfunction, and male infertility. Precisely in Northeast Brazil, ovine consist of the leading livestock produced, and their reproduction in captivity has been demonstrated difficult. Thus, we hypothesized whether the application of letrozole will improve male sheep reproduction. One group of 6 animals received a daily dosage of 0.5mg/Kg of letrozole for 60 days, while the other six animals were used as control. Samples were collected from control and treated animals after 30 and 60 days of the experiment. Blood samples were collected to measure the steroid hormone levels. Semen was collected from control and treated groups using an artificial vagina and cryopreserved for spermatozoa morphology and CASA analysis. The testicles were collected for histological analysis, gene expression, and immunohistochemistry of P450aromatase protein. Hormone levels demonstrate no differences in the estradiol/testosterone levels among the control and both treated groups. Immunohistochemistry analysis revealed the presence of P450aromatase protein in spermatogonia cells and Leydig cells in the control and treated groups in both periods analyzed. Moreover, there were no differences in the P450aromase gene expression in the control and treated group. Morphological analysis of the spermatozoa revealed that letrozole treatment did not affect mitochondrial activity or cause any deformities. In addition, motility parameters in the sperm from the treated group were not affected by letrozole treatment compared to the control group. Morphological analysis of the testis demonstrated that letrozole treatment increase the seminiferous tubule area but no signs of germ cell damage. Our results show that letrozole has a morphological effect on the testicles in the ovine model but no pathological or severe effect caused by hormone level imbalance. Overall, letrozole comprises a non-steroid aromatase inhibitor, which can be applied to ovine reproduction. nonsteroid non uses prevention dysfunction infertility Brazil produced difficult Thus 05mgKg mgKg 0 5mg Kg mg 3 experiment Paromatase P estradioltestosterone estradiol testosterone analyzed Moreover Paromase aromase deformities addition damage imbalance Overall

Abstract This study aimed to investigate the effect of growth and differentiation factor 9 (GDF-9) during the in vitro culture of isolated caprine early antral follicles. The isolated and selected early antral follicles were individually cultured for 18 days, and the following treatments were tested: α-MEM+ (control treatment) or α-MEM+ supplemented with 200 ng/mL GDF-9. The following endpoints were evaluated: follicular growth and morphology, estradiol production, oocyte nuclear maturation, and relative expression of key genes related to steroidogenesis (CYP19A1, CYP17, and insulin receptor) and basement membrane remodeling (MMP-9 and TIMP-2). In both treatments, a decrease was observed in the percentage of morphologically intact follicles with a concomitant increase in the rates of extruded and degenerated follicles (P < 0.05). The GDF-9 treatment showed higher rates of extruded follicles only on day 6 of culture (P < 0.05). Follicle diameter increased progressively throughout the culture period (P < 0.05) with similar diameters between treatments at all culture times (P > 0.05). Growth and differentiation factor 9 increased the daily growth rate from the first to the second third of culture, with higher values (P < 0.05) than control in the second third. Oocyte maturation rate as well as estradiol levels and relative mRNA expression for CYP19A1, CYP17, MMP-9, TIMP-2, and insulin receptor genes were similar between treatments (P > 0.05). This study shows for the first time that GDF-9 added to a culture medium increased the follicle growth rate of goat early antral follicles cultured in vitro. GDF9 GDF (GDF-9 1 days tested αMEM+ αMEM α MEM+ MEM α-MEM 20 ngmL ng mL GDF9. 9. evaluated morphology production CYP19A1 CYPA CYP A (CYP19A1 CYP17 MMP9 MMP (MMP- TIMP2. TIMP2 TIMP 2 . TIMP-2) P 0.05. 005 0.05 0 05 GDF- MMP9, 9, MMP-9 TIMP2, 2, TIMP-2 (GDF- CYP19A (CYP19A CYP1 (MMP 00 0.0 MMP- TIMP- (GDF 0.

Resumo Este estudo teve como objetivo investigar o efeito do GDF-9 durante o cultivo in vitro de folículos antrais iniciais caprinos isolados. Os folículos antrais iniciais isolados e selecionados foram cultivados individualmente por 18 dias, e os seguintes tratamentos foram testados: α MEM+ (tratamento controle) ou α-MEM+ suplementado com 200 ng/mL de GDF-9 (tratamento GDF-9). Os seguintes parâmetros foram avaliados: crescimento e morfologia folicular, produção de estradiol, maturação nuclear do oócito e expressão relativa de genes-chave relacionados a esteroidogênese (CYP19A1, CYP17 e receptor de insulina) e remodelamento da membrana basal (MMP-9 e TIMP-2). Em ambos os tratamentos, observou-se diminuição na porcentagem de folículos morfologicamente intactos com aumento concomitante nas taxas de folículos extrusos e degenerados (P < 0,05). O tratamento GDF-9 apresentou maiores taxas de folículos extrusos apenas no 6º dia de cultivo (P < 0,05). O diâmetro do folículo aumentou progressivamente ao longo do período de cultivo (P < 0,05) com diâmetros semelhantes entre os tratamentos em todos os tempos de cultivo (P > 0,05). O GDF-9 aumentou a taxa de crescimento diário do primeiro para o segundo terço de cultivo, sendo maior (P < 0,05) que o controle no segundo terço. A taxa de maturação oocitária assim como os níveis de estradiol e a expressão relativa de RNAm para os genes CYP19A1, CYP17, MMP-9, TIMP-2 e receptor de insulina foram similares entre os tratamentos (P > 0,05). Em conclusão, este estudo mostra pela primeira vez que GDF-9 adicionado a um meio de cultivo aumentou a taxa de crescimento de folículos antrais iniciais caprinos cultivados in vitro. GDF9 GDF 9 GDF- 1 dias testados MEM αMEM+ αMEM α-MEM 20 ngmL ng mL GDF9. . GDF-9) avaliados folicular geneschave chave CYP19A1 CYPA CYP (CYP19A1 CYP1 MMP9 MMP (MMP- TIMP2. TIMP2 TIMP 2 TIMP-2) observouse observou se P 0,05. 005 0,05 0 05 º MMP9, 9, MMP-9 TIMP- conclusão CYP19A (CYP19A (MMP 00 0,0 MMP- 0,

ABSTRACT Adrenal steroid biosynthesis and its related pathology are constant evolving disciplines. In this paper, we review classic and current concepts of adrenal steroidogenesis, plus control mechanisms of steroid pathways, distribution of unique enzymes and cofactors, and major steroid families. We highlight the presence of a “mineralocorticoid (MC) pathway of zona fasciculata (ZF)”, where most circulating corticosterone and deoxycorticosterone (DOC) originate together with 18OHDOC, under ACTH control, a claim based on functional studies in normal subjects and in patients with 11β-, and 17α-hydroxylase deficiencies. We emphasize key differences between CYP11B1 (11β-hydroxylase) and CYP11B2 (aldosterone synthase) and the onset of a hybrid enzyme – CYP11B1/CYP11B2 –, responsible for aldosterone formation in ZF under ACTH control, in “type I familial hyperaldosteronism” (dexamethasone suppressible). In “apparent MC excess syndrome”, peripheral conversion of cortisol to cortisone is impaired by lack of 11β-hydroxysteroid dehydrogenase type 2, permitting free cortisol access to MC receptors resulting in severe hypertension. We discuss two novel conditions involving the synthesis of adrenal androgens: the “backdoor pathway”, through which dihydrotestosterone is formed directly from androsterone, being relevant for the fetoplacental setting and sexual differentiation of male fetuses, and the rediscovery of C19 11-oxygenated steroids (11-hydroxyandrostenedione and 11-ketotestosterone), active androgens and important markers of virilization in 21-hydroxylase deficiency and polycystic ovaries syndrome. Finally, we underline two enzyme cofactor deficiencies: cytochrome P450 oxidoreductase which partially affects 21- and 17α-hydroxylation, producing a combined clinical/hormonal picture and causing typical skeletal malformations (Antley-Bixler syndrome), and PAPSS2, coupled to SULT2A1, that promotes sulfation of DHEA to DHEAS, preventing active androgens to accumulate. Its deficiency results in reduced DHEAS and elevated DHEA and androgens with virilization. Future and necessary studies will shed light on remaining issues and questions on adrenal steroidogenesis.

Abstract Arsenic exposure is a global health concern. This toxic metalloid is ubiquitous in the environment and contaminates food and drinking water. Once ingested, it undergoes a complex metabolic process within the body, which contributes to its accumulation and reactivity. Arsenic toxicity stems from the induction of oxidative stress, inhibition of thiol-containing proteins, and mimicry of inorganic phosphates. Arsenic poisoning is associated with the development of reproductive disorders. In males, arsenic causes a reduction in testicular weight and alterations in steroidogenesis and spermatogenesis. Moreover, it reduces the number and quality of spermatozoa harvested from the cauda epididymis. The mitochondria are targets of arsenic toxicity because of the production of free radicals and their high content of cysteine-rich proteins and fatty acids. Mitochondrial dysfunction may contribute to reproductive disorders because this organelle is crucial for controlling testicular and epididymal events related to sperm production and maturation. All of these alterations mediated by arsenic exposure contribute to the failure of male reproductive competence by reducing gamete viability. This review describes the potential mechanisms of arsenic toxicity, its detrimental effects on male reproductive organs, and consequences on sperm fertility. concern water ingested body reactivity stress thiolcontaining thiol containing phosphates males spermatogenesis Moreover epididymis cysteinerich cysteine rich acids maturation viability organs fertility

Abstract The relationship between endometriosis and infertility is still unknown, but it is possible that genetic polymorphisms influence these two variables. This study aims to identify, in the literature, which polymorphisms are related to infertility in women with endometriosis. A search was performed in databases using the descriptors: polymorphisms genetics and infertility and endometriosis. 386 articles were identified, and after applying the inclusion and exclusion criteria, 33 case-control studies were included. Genes and their respective polymorphisms, which exhibited statistically significant values, were classified into three categories: related to metabolic/cellular processes, steroidogenesis and sex hormone receptors, inflammation and immune response. In summary, the results of these studies suggest that the polymorphisms rs882605 of MUC4 gene, rs16826658 of WNT4 gene, rs10953316 of MUC17 gene, rs10928050 of KAZN gene, rs1799889 of PAI-1 gene, (TA)n repeats of ESR1 gene, (CA)n repeats of ESR2 gene, rs605059 of HSD17B1 gene, rs743572 of CYP17A1 gene, insLQ of LHR gene, p.Ile49Ser of AMH gene, rs12700667 of NPVF/NFE2L3 gene, G1502A of LHβ gene, G + 1730A of ERβ gene, rs7528684 of FCRL3 gene, rs3761549 of FOXP3 gene and rs28362491 of NFKβ1 gene are implicated in the etiology of infertility in women with endometriosis.

Summary: Growth hormone is recognized as a key endocrine regulator of postnatal growth and production in dairy cattle. However, it is currently known that this hormone also affects reproduction in these animals. Ovarian function in cattle is controlled by intraovarian control mechanisms that, concomitantly with endocrine signals, coordinate recruitment, selection and follicular growth. Although the role of gonadotropins in stimulating follicular growth and differentiation is clear, particularly during the final stages of maturation, growth hormone also plays an important modulating role, since it exerts multiple direct and indirect effects on the ovary, acting on steroidogenesis, folliculogenesis and oocyte maturation. Any advances in understanding intraovarian regulatory mechanisms will be helpful in the design of new approaches to manipulating ovarian function andimproving fertility in dairy herds.

RESUMEN: La hormona del crecimiento es reconocida como un regulador endocrino clave del crecimiento postnatal y la producción en bovinos de leche. No obstante, actualmente se conoce que esta hormona también afecta la reproducción de estos animales. La función ovárica en los bovinos está controlada por mecanismos de control intraováricos que, concomitantemente con señales endócrinas, coordinan el reclutamiento, la selección y el crecimiento folicular. Aunque el rol de las gonadotropinas en la estimulación del crecimiento y diferenciación folicular es claro, particularmente durante las etapas finales de la maduración, la hormona del crecimiento también cumple una función moduladora de relevancia, ya que ejerce múltiples efectos directos e indirectos en el ovario, actuando sobre la esteroidogénesis, la foliculogénesis y la maduración de los ovocitos. Todo avance en la comprensión de los mecanismos de regulación intraováricos resultará de utilidad en la planificación de nuevos enfoques para la manipulación de la función ovárica y la mejora de la fertilidad de los rodeos lecheros.

Abstract Ginger is traditionally used as a sexual enhancer in folk medicine. Despite extensive studies on the effect of ginger on reproduction, the molecular mechanism of ginger prevention effect on ethanol-induced reproductive disorder is not fully understood. Twenty-four adult male ratswereallocated into control, ethanol (4 g/kg of body weight (BW)/day), ginger (250 mg/kg of BW/day) and ginger-ethanol group. Ginger and ethanol were administrated by gavage for 28 days. Testicular concentration of testosterone, TNF-α, and antioxidant enzymes activity and serum concentration of gonadotropins hormone and testosterone were measured. The gene expression of Nrf2 and NF-κB which regulate oxidative damage and inflammation, respectively, and StAR, P450scc and 17βHSD which are involved in testosterone synthesis were detected. Ethanol significantly decreased gonadotropin hormones, oxidative markers, expression of genes involved in testosterone synthesis and Nrf2, and in reverse significantly increased TNF-α, MDA and gene expression of NF-κB compared to control (p<0.05). While ginger could significantly improve all of the above factors compared to the ethanol group (p<0.05). These results were also supported by histological findings. It can be concluded that ginger prevents the ethanol-induced reproductive dysfunction by improving the gonadotropins, oxidative damage and inflammation and the genes involved in testosterone synthesis.

Abstract Letrozole is used as a therapeutic agent in reproductive disorders caused by high estrogen levels. Letrozole inhibits cytochrome P450 aromatase and reduces estrogen levels. However, the effects of long-term use on reproductive traits are unknown. The aim of this study was to evaluate the prolonged use of letrozole in the gonads of rodents (Spix's yellow-toothed cavy; Galea spixii). Forty-eight rodents (24 males and 24 females) were randomly divided into the treated and control groups. Letrozole administration started at 15 days of age and continued weekly until 30, 45, 90, and 120 days of age. The body, testis, and ovary weights were analyzed, as well as the morphological progression of spermatogenesis and folliculogenesis. Macroscopically, body weight gain and gonads weight were increased in the letrozole group. Microscopically, the ovaries of treated females showed stratified epithelium and a cellular disorder of the tunica albuginea. In the testes of treated males, the development of seminiferous tubules was delayed and sperm was absent. The collective findings indicate that the prolonged use of letrozole alters secondary sexual characteristics, and causes weight gain, reproductive changes, and male infertility.

ABSTRACT Objective To analyze the morphological and functional characteristics of primary macronodular adrenal hyperplasia (PMAH) nodules carrying or not carrying ARMC5 mutations and the consequences of the presence of mutations in terms of the pattern of macronodule composition and functional state. Subjects and methods The analyses were performed by hematoxylin-eosin staining, immunohistochemistry, microdissection of spongiocyte tissue and RT-qPCR of histological sections from 16 patients diagnosed with PMAH with germline (5) or germline/somatic mutations (5) and without mutations (6) in the ARMC5 gene. Results Hyperplastic nodules were predominantly composed of spongiocytes in mutated and nonmutated sections. ARMC5 mRNA expression in spongiocytes was higher in ARMC5-mutated nodules than in ARMC5-nonmutated nodules, and homogenous ARMC5 protein distribution was observed. The presence of arginine-vasopressin receptor (AVP1AR) and ectopic ACTH production were observed in both cell populations regardless of ARMC5 mutations; the numbers of serotonin receptor (5HT4R)- and proliferating cell nuclear antigen (PCNA)-positive cells were higher in macronodules carrying ARMC5 mutations than in those without mutations. Conclusions Our results suggest that the presence of ARMC5 mutations does not interfere with the pattern of distribution of spongiocytes and compact cells or with the presence of AVP1AR, gastric-inhibitory polypeptide receptor (GIPR) and ectopic ACTH. Nevertheless, the higher numbers of PCNA-positive cells in mutated nodules than in nonmutated nodules suggest that mutated ARMC5 can be related to higher proliferation rates in these cells. In conclusion, our results provide more information about the crosstalk among abnormal GPCRs, ectopic ACTH in steroidogenesis and the ARMC5 gene, which may be relevant in understanding the pathogenesis and diagnosis of patients with PMAH.

Abstract The aim of this study was to evaluate the long-term effects of recombinant bovine somatotropin (rbST) on follicle population and ovulatory follicle development in non-lactating dairy cows. Twenty-one Jersey cows were allocated in rbST (n=11) or control (n=10) groups. On day -60, cows in rbST group received 500 mg of somatotropin (s.c. Lactotropin®, Elanco). On day 0, control and rbST cows received an intravaginal progesterone-releasing device (1.9 g, CIDR®, Zoetis) and GnRH (100 mg, IM, Factrel®, Zoetis). On day 8, cows received PGF2α (25 mg, IM, Lutalyse®, Zoetis) and the CIDR® was removed. Twelve hours after device removal (D8), serum, follicular fluid and granulosa cells samples were collected. Serum and follicular concentration of estradiol (E2) and progesterone (P4) were analyzed. Total RNA was extracted from granulosa cells to measure gene expression of LHCGR, STAR, HSD-3B1, CYP11A1, CYP19A1, CYP17A1, IGFR and PAPPA by real-time PCR. Ultrasonography was performed on days -60, -53, -46, -14, -7, 0 and 8 for antral follicle count. Results were analyzed by repeated measures ANOVA and t-test. There was no effect of rbST treatment on the number of follicles during the 60 days period, as well as no effect on serum and follicular fluid E2 and follicular fluid P4 at the moment of follicle aspiration. There was a reduction in PAPPA (P = 0.006), CYP11A1 (P = 0.04) and CYP19A1 (P = 0.002) mRNA levels in granulosa cells of the pre-ovulatory follicle of rbST treated cows. In conclusion, a single dose of rbST did not have long-term effects on antral follicle population, serum and follicular E2/P4 concentrations in non-lactating dairy cows. Despite that, rbST injection decreased granulosa cell expression of genes related to steroidogenesis in the pre-ovulatory follicle.