Dr Association of bone morphogenetic protein 15 and growth differentiation factor 9 with litter size in livestock: a review study

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 ß(TGF-ß) superfamily includes more than 34 members; GDF9 and BMP15 are among the most significantfactors 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 reviewprovides valuable information on the association of BMP15 and GDF9with litter size in livestock that can be used as biological markers of multiple ovulation or for improving fertility in livestock.(AU)

Morphology and immunolocalization of intertubular steroidogenic cell in mesonephros of Podocnemis expansa during gonadal differentiation

Sex steroid hormones are critical in gonadal differentiation in turtles. The gonads are not the only organs responsible for producing these hormones during this phase. Mesonephros play an important role in steroidogenesis. The present study aimed to investigate the presence of steroidogenic cells in mesonephros of Podocnemis expansa during gonadal differentiation and to evaluate their morphology and ultrastructure. Ten embryos of P. expansa were collected from 5 nests on day 36 of incubation, during spawning period on an artificial beach. Embryos were extracted from eggs by slicing the shell and euthanized. They were dissected under a stereoscopic microscope to collect the gonad-mesonephro complex, in which were fixed and subsequently processed for light microscopy, immunohistochemistry and transmission electron microscopy analysis. During histological analysis was observed mesonephros has typical morphological structure. Immunohistochemistry showed immunoreaction to aromatase in cells of intertubular space. Confirming these findings, it was possible to observe a type of intertubular cell in several regions of mesonephro, being more predominant in region close to blood vessels, distal and proximal tubules. In ultrastructural analysis these cells were characterized by having a clear, large, and rounded nucleus with evident nucleolus and cytoplasm rich in electron-dense droplets. This study demonstrated for the first time the presence of cells with morphological, immunohistochemical and ultrastructural characteristics similar to steroid-producing cells in P. expansa mesonephrons, suggesting that this organ may contribute to gonadal differentiation in this species.(AU)

Arsenic exposure and its implications in male fertility

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.(AU)

Prolonged use of letrozole causes morphological changes on gonads in Galea spixii

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.

Prolonged use of letrozole causes morphological changes on gonads in Galea spixii

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 longterm 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.(AU)

Doenças do pós-parto e seus efeitos sobre a eficiência reprodutiva de vacas leiteiras / Postpartum diseases and their effects on dairy cows reproductive efficiency

O período de transição das vacas leiteiras é marcado por diversas alterações fisiológicas, metabólicas e endócrinas. Essas alterações aumentam a susceptibilidade das vacas às infecções no pósparto o que compromete bem-estar, resultando em menor desempenho produtivo e reprodutivo, levando ao aumento dos custos relacionados ao tratamento das infecções e ao descarte de leite. Dentre as doenças que acometem as vacas leiteiras no período pós-parto destacam-se as patologias relacionadas ao trato reprodutivo (retenção de placenta, metrite, endometrite clínica e subclínica) ou em outros órgãos (mastite, claudicação, problemas digestivos e respiratórios), e os distúrbios metabólicos (cetose, hipocalcemia e deslocamento de abomaso). Embora o impacto negativo das doenças no pós-parto sobre a eficiência reprodutiva de vacas leiteiras já tenha sido demonstrado por diversos estudos, os mecanismos pelos quais os processos infecciosos e inflamatórios interferem direta e/ou indiretamente no funcionamento do trato reprodutivo das vacas leiteiras ainda não foram totalmente elucidados. Sabe-se que as respostas inflamatória e imunológica frente à infecção podem influenciar processos reprodutivos como a dinâmica folicular ovariana, a esteroidogênese, a competência ovocitária, a fertilização, o desenvolvimento embrionário e a manutenção da gestação. Portanto, a ocorrência de doenças no pós-parto é um obstáculo a eficiência reprodutiva, principalmente devido à alta incidência dessas infecções nas vacas de leite. Podese concluir que a prevenção é a melhor estratégia para combater os efeitos negativos da ocorrência das doenças do pós-parto na eficiência reprodutiva, pois mesmo após a cura clínica dessas patologias a eficiência reprodutiva não é reestabelecida, e seus efeitos persistem por toda a lactação.

The transition period of dairy cows is marked by several physiological, metabolic and endocrine changes. These changes increase the susceptibility of cows to postpartum infections, which compromises welfare, resulting in lower productive and reproductive performance, leading to increased costs related to the treatment of infections and milk disposal. Among the diseases that affect dairy cows in the postpartum period are pathologies related to the reproductive tract (retention of placenta, metritis, clinical and subclinical endometritis) or to other organs (mastitis, claudication, digestive, and respiratory problems), and metabolic disorders (ketosis, hypocalcemia and abomasal displacement). Although the negative impact of postpartum diseases on dairy cows reproductive efficiency has already been demonstrated by several studies, the mechanisms by which infectious and inflammatory processes directly and/or indirectly interfere in the reproductive tract of dairy cows have not yet been fully elucidated. It is known that inflammatory and immunological responses to infection can influence reproductive processes such as ovarian follicular dynamics, steroidogenesis, oocyte competence, fertilization, embryonic development, and maintenance of pregnancy. Therefore, the occurrence of postpartum diseases is an obstacle to reproductive efficiency, mainly due to the high incidence of these infections in dairy cows. It can be concluded that prevention is the best strategy to combat the negative effects of the occurrence of postpartum diseases on reproductive efficiency, because even after the clinical cure of these pathologies reproductive efficiency is not reestablished, and its effects persist throughout lactation.

Hormônio antimülleriano como marcador de fertilidade em fêmeas suínas / Anti-Müllerian hormone as a fertility marker in swine females

O hormônio antimülleriano é conhecido por seu papel na regressão dos ductos de Müller durante a fase de diferenciação sexual dos machos. Ainda assim, esse hormônio atua de forma ainda mais ampla no desenvolvimento de ambos os sexos. O hormônio antimülleriano é diferencialmente expresso em machos e fêmeas durante toda a vida. Nas fêmeas esse hormônio é produzido pelas células da granulosa dos folículos ovarianos e atua modulando os processos de secreção de gonadotrofinas, foliculogênese e esteroidogênese, sendo considerado um biomarcador confiável para fertilidade em diferentes espécies. Apesar de pouquíssimo estudado na espécie suína, alguns trabalhos vêm demonstrando a existência de associações entre os níveis séricos do hormônio antimülleriano e os fenótipos de maior fertilidade. De forma que, o objetivo desta revisão é fazer um levantamento do conhecimento já acumulado acerca da temática, demostrando a importância do desenvolvimento de novos estudos; tendo em vista que o aprofundamento do conhecimento sobre as funções do hormônio antimülleriano nas fêmeas da espécie suína pode contribuir para o melhor entendimento da fisiologia da espécie(AU)

The anti-Müllerian hormone (AMH) is known for its role in the regression of Muller ducts during the phase of sexual differentiation in males. Notwithstanding, this hormone acts more broadly on the development of both sexes. The AMH is differentially expressed both in males and females throughout life. In females, this hormone is produced by granulosa cells of ovarian follicles and modulates the gonadotropin secretion, folliculogenesis, and steroidogenesis, and is considered a reliable fertility biomarker in different species. Despite the lack of studies in the swine species, some reports demonstrated the association between serum levels of AMH and more fertile phenotypes. Therefore, this review aims to survey the current knowledge about this theme, demonstrating the importance of the development of new research, since the strengthening of the knowledge on the functions of the AMH in swine females may contribute to understanding the physiology of the species.(AU)

Hormônio antimülleriano como marcador de fertilidade em fêmeas suínas / Anti-Müllerian hormone as a fertility marker in swine females

O hormônio antimülleriano é conhecido por seu papel na regressão dos ductos de Müller durante a fase de diferenciação sexual dos machos. Ainda assim, esse hormônio atua de forma ainda mais ampla no desenvolvimento de ambos os sexos. O hormônio antimülleriano é diferencialmente expresso em machos e fêmeas durante toda a vida. Nas fêmeas esse hormônio é produzido pelas células da granulosa dos folículos ovarianos e atua modulando os processos de secreção de gonadotrofinas, foliculogênese e esteroidogênese, sendo considerado um biomarcador confiável para fertilidade em diferentes espécies. Apesar de pouquíssimo estudado na espécie suína, alguns trabalhos vêm demonstrando a existência de associações entre os níveis séricos do hormônio antimülleriano e os fenótipos de maior fertilidade. De forma que, o objetivo desta revisão é fazer um levantamento do conhecimento já acumulado acerca da temática, demostrando a importância do desenvolvimento de novos estudos; tendo em vista que o aprofundamento do conhecimento sobre as funções do hormônio antimülleriano nas fêmeas da espécie suína pode contribuir para o melhor entendimento da fisiologia da espécie

The anti-Müllerian hormone (AMH) is known for its role in the regression of Muller ducts during the phase of sexual differentiation in males. Notwithstanding, this hormone acts more broadly on the development of both sexes. The AMH is differentially expressed both in males and females throughout life. In females, this hormone is produced by granulosa cells of ovarian follicles and modulates the gonadotropin secretion, folliculogenesis, and steroidogenesis, and is considered a reliable fertility biomarker in different species. Despite the lack of studies in the swine species, some reports demonstrated the association between serum levels of AMH and more fertile phenotypes. Therefore, this review aims to survey the current knowledge about this theme, demonstrating the importance of the development of new research, since the strengthening of the knowledge on the functions of the AMH in swine females may contribute to understanding the physiology of the species.

Impact of hypotensive action of angiotensin converting enzyme inhibitor on ovarian-intraovarian blood flow and follicles development in goats hormonally stimulated with repeated FSH-ECG treatment

Background: Recent evidence shows that the renin-angiotensin system (RAS) participates in important reproductiveprocesses, such as steroidogenesis, folliculogenesis, oocyte maturation and ovulation. Several studies have proposed touse an angiotensin-converting enzyme (ACE) as a RAS modulator, aiming to improve reproductive efficiency, however,the presence of the main components of this system in reproductive tissues still need to be further investigated, since thephysiological functions seem to be species-specific. The aim of this study was to assess the impact of enalapril-maleate,an ACE inhibitor, during repeated gonadotropins treatment on ovarian blood flow and follicular development in goats.Materials, Methods & Results: Twenty Anglo-Nubian cross-bred goats were equally grouped according to parity (n= 10/group): nulliparous and multiparous parity. In each group, five animals were randomly selected to receive 0.4 mg.kg-1 ofenalapril-maleate during 11 days of estrus synchronization and gonadotropins treatments. The other animals received thesame volume of saline solution. Estrus synchronization of all goats was made by intramuscular ad-ministration of PGF2αanalog, followed 48 h later by intravaginal insertion of a controlled internal drug release device. Forty-eight h after devicewithdrawal, a single dose of 60 mg of FSH plus 300 UI of eCG was administered and repeated every 4 days to complete 3treatments. Transrectal ultrasonography was performed using pulsed and color Doppler to evaluate Doppler velocimetricsparameters of the ovarian artery and intraovarian blood flow, respec-tively, and B-mode real-time ultrasound scanner toevaluate the follicular development. In the females treated with enalapril-maleate was observed a significant reduction ofsystolic and diastolic peak, without difference according to parity. In addition, in the third session of hor-monal stimulation,only the groups (nulliparous and multiparous) not treated with...(AU)

Impact of hypotensive action of angiotensin converting enzyme inhibitor on ovarian-intraovarian blood flow and follicles development in goats hormonally stimulated with repeated FSH-ECG treatment

Background: Recent evidence shows that the renin-angiotensin system (RAS) participates in important reproductiveprocesses, such as steroidogenesis, folliculogenesis, oocyte maturation and ovulation. Several studies have proposed touse an angiotensin-converting enzyme (ACE) as a RAS modulator, aiming to improve reproductive efficiency, however,the presence of the main components of this system in reproductive tissues still need to be further investigated, since thephysiological functions seem to be species-specific. The aim of this study was to assess the impact of enalapril-maleate,an ACE inhibitor, during repeated gonadotropins treatment on ovarian blood flow and follicular development in goats.Materials, Methods & Results: Twenty Anglo-Nubian cross-bred goats were equally grouped according to parity (n= 10/group): nulliparous and multiparous parity. In each group, five animals were randomly selected to receive 0.4 mg.kg-1 ofenalapril-maleate during 11 days of estrus synchronization and gonadotropins treatments. The other animals received thesame volume of saline solution. Estrus synchronization of all goats was made by intramuscular ad-ministration of PGF2αanalog, followed 48 h later by intravaginal insertion of a controlled internal drug release device. Forty-eight h after devicewithdrawal, a single dose of 60 mg of FSH plus 300 UI of eCG was administered and repeated every 4 days to complete 3treatments. Transrectal ultrasonography was performed using pulsed and color Doppler to evaluate Doppler velocimetricsparameters of the ovarian artery and intraovarian blood flow, respec-tively, and B-mode real-time ultrasound scanner toevaluate the follicular development. In the females treated with enalapril-maleate was observed a significant reduction ofsystolic and diastolic peak, without difference according to parity. In addition, in the third session of hor-monal stimulation,only the groups (nulliparous and multiparous) not treated with...

Effect of recombinant bovine somatotropin (rbST) treatment on follicular population and development in non-lactating dairy cows

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.

Effect of recombinant bovine somatotropin (rbST) treatment on follicular population and development in non-lactating dairy cows

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.(AU)

The local regulation of folliculogenesis by members of the transforming growth factor superfamily and its relevance for advanced breeding programmes

Regulation of the growth and maturation of the ovarian follicle is critical for normal reproductive function. Alterations in this growth can lead to pathological conditions, such as cystic follicles, reduced oocyte quality, or an abnormal endocrine environment leading to poor fertility. Alterations in follicular growth also influence the number of follicles ovulating and thus can change litter size. Both endocrine factors, such a sfollicle stimulating hormone and luteinizing hormone, as well as local factors, are known to regulate follicular growth and development. This review will focus on the role of local factors in regulation of ovarian follicular growth in ruminants, with a focus on members of the transforming growth factor superfamily. The potential role of these factors in regulating proliferation, apoptosis, steroidogenesis and responsiveness to gonadotrophins will be considered.

The local regulation of folliculogenesis by members of the transforming growth factor superfamily and its relevance for advanced breeding programmes

Regulation of the growth and maturation of the ovarian follicle is critical for normal reproductive function. Alterations in this growth can lead to pathological conditions, such as cystic follicles, reduced oocyte quality, or an abnormal endocrine environment leading to poor fertility. Alterations in follicular growth also influence the number of follicles ovulating and thus can change litter size. Both endocrine factors, such a sfollicle stimulating hormone and luteinizing hormone, as well as local factors, are known to regulate follicular growth and development. This review will focus on the role of local factors in regulation of ovarian follicular growth in ruminants, with a focus on members of the transforming growth factor superfamily. The potential role of these factors in regulating proliferation, apoptosis, steroidogenesis and responsiveness to gonadotrophins will be considered.(AU)

Simazine, a triazine herbicide, disrupts swine granulosa cell functions

The triazine herbicide simazine is a pesticide commonly detected in surface and ground waters, although banned in most European countries since 2004. Concerns for humans and animal health result from its potential endocrine disrupting action, that can lead to reproductive disorders. The present in vitro study was undertaken to study simazine effects on swine granulosa cell function, namely cell viability, proliferation, steroidogenesis and NO production. Moreover, the ability of this substance to interfere with the angiogenetic process, a crucial event in reproductive function, was taken into account. Our data document that simazine treatment, at 0.1 or 10 μM concentration levels, stimulates granulosa cell proliferation and viability and impairs steroidogenesis, increasing in particular progesterone production. In addition, the in vitro angiogenesis bioassay revealed a significant simazine stimulatory effect on immortalized porcine Aortic Endothelial Cell proliferation. Collectively, these results show that simazine can display disruptive effects on ovarian cell functional parameters, possibly resulting in reproductive dysfunction. This hypothesis is also supported by the observed pro-angiogenetic properties of this herbicide, as already suggested for different endocrine disruptors.

Simazine, a triazine herbicide, disrupts swine granulosa cell functions

The triazine herbicide simazine is a pesticide commonly detected in surface and ground waters, although banned in most European countries since 2004. Concerns for humans and animal health result from its potential endocrine disrupting action, that can lead to reproductive disorders. The present in vitro study was undertaken to study simazine effects on swine granulosa cell function, namely cell viability, proliferation, steroidogenesis and NO production. Moreover, the ability of this substance to interfere with the angiogenetic process, a crucial event in reproductive function, was taken into account. Our data document that simazine treatment, at 0.1 or 10 μM concentration levels, stimulates granulosa cell proliferation and viability and impairs steroidogenesis, increasing in particular progesterone production. In addition, the in vitro angiogenesis bioassay revealed a significant simazine stimulatory effect on immortalized porcine Aortic Endothelial Cell proliferation. Collectively, these results show that simazine can display disruptive effects on ovarian cell functional parameters, possibly resulting in reproductive dysfunction. This hypothesis is also supported by the observed pro-angiogenetic properties of this herbicide, as already suggested for different endocrine disruptors.(AU)

Granulosa cell tumor associated with occult hyperadrenocorticism in a yorkshire terrier bitch

Background: Hyperadrenocorticism (HAC), is considered a set of symptoms due to excessive exposure to cortisol. Naturally occurring HAC is most often related to pituitary tumors (pituitary-dependent HAC - PDH). Occult HAC, is referred as a clinical picture highly consistent with HAC; however, routine screening tests are negative. In addition, one or more steroids are elevated following administration of adrenocorticotrophic hormone (ACTH). Ovarian granulosa cell tumors, can produce steroids leading to paraneoplastic syndromes. The objective of this study was to report an unpublished case of ovarian granulosa cell tumor associated with occult hyperadrenocorticism in a Yorkshire Terrier.Case: A 13-year-old intact female dog, Yorkshire Terrier, was brought for consultation with slight weight loss, polyuria, polydipsia, irregular estrous cycles, increased abdominal volume and thin coat. On physical examination the animal was gasping and presented severe periodontal disease, bulging abdomen, alopecia and skin hyperpigmentation. Complete blood count presented no changes; however, serum biochemistry evaluation highlighted hyperalbuminemia, hypertriglyceridemia, alkaline phosphatase increased activity, and urinary specific gravity and creatinine below reference values. On abdominal ultrasonography left adrenal gland measured 2.08 cm x 1.08 cm and the right adrenal gland measured 2.11 cm x 0.84 cm, indicating bilateral adrenomegaly compatible with PDH. In the hypogastric abdomen, a large heterogeneous hypoechogenic mass was also observed, with areas of cystic cavities, measuring 5.80 cm x 7.30 cm. A low dose dexamethasone suppression test (LDDST) was performed, due to PDH suspicion. The test did not confirm HAC, suspecting, then, to be a case of occult/atypical HAC. Due to the strong clinical suspicion, and owner financial problems for further investigated occult HAC, trilostane treatment was initiated.[...](AU)

Parthenogenetic bovine embryos secrete type I interferon capable of stimulating ISG15 in luteal cell culture

Interferon tau (IFNT) is the pregnancy recognition signal in ruminants and is secreted by trophoblast cells. Paracrine action in the endometrium is well established by inhibiting luteolytic pulses of prostaglandin F2 alpha. Recently, endocrine action was documented in the corpus luteum, blood cell and liver. It was hypothesized that conditioned medium (CM) obtained from days 7, 9 and 12 parthenogenetic embryos alters luteal cell gene expression. The aim was to establish a bovine mixed luteal cell culture to evaluate cellular response associated to interferon stimulated genes, steroidogenesis and apoptosis. Conditioned medium was obtained from Days 7, 9 and 12 parthenogenetic (PA) embryos culture. Moreover, antiviral assay was performed on CM from Days 7, 9 and 12 to verify Type I interferon activity. Luteal cell culture was validated by steroidogenic and apoptotic genes (CYP11A1, HSD3B1, BAX, BCL2, AKT and XIAP mRNA expression), and concentration of progesterone as endpoint. Luteal cell culture was treated with interferon alpha (IFNA) and CM from parthenogenetic embryos. Antiviral assay revealed Type I interferon activity on CM from embryos increasing on Days 9 and 12. ISG15 mRNA was greater in the mixed luteal cells culture treated with 1, 10 and 100ng/ml of interferon alpha (IFNA) and also on Days 7, 9 and 12 CM treatments. Concentration of progesterone was not altered in luteal cell culture regardless of treatments. Steroidogenic and apoptotic genes were similar among groups in luteal cell culture treated with different doses of IFNA or CM from PA embryos. In conclusion, parthenogenetic embryo-derived CM has antiviral activity, luteal cell culture respond to Type I interferon by expressing IGS15. These data indicate this model can be used for IFNT endocrine signaling studies.

Granulosa cell tumor associated with occult hyperadrenocorticism in a yorkshire terrier bitch

Background: Hyperadrenocorticism (HAC), is considered a set of symptoms due to excessive exposure to cortisol. Naturally occurring HAC is most often related to pituitary tumors (pituitary-dependent HAC - PDH). Occult HAC, is referred as a clinical picture highly consistent with HAC; however, routine screening tests are negative. In addition, one or more steroids are elevated following administration of adrenocorticotrophic hormone (ACTH). Ovarian granulosa cell tumors, can produce steroids leading to paraneoplastic syndromes. The objective of this study was to report an unpublished case of ovarian granulosa cell tumor associated with occult hyperadrenocorticism in a Yorkshire Terrier.Case: A 13-year-old intact female dog, Yorkshire Terrier, was brought for consultation with slight weight loss, polyuria, polydipsia, irregular estrous cycles, increased abdominal volume and thin coat. On physical examination the animal was gasping and presented severe periodontal disease, bulging abdomen, alopecia and skin hyperpigmentation. Complete blood count presented no changes; however, serum biochemistry evaluation highlighted hyperalbuminemia, hypertriglyceridemia, alkaline phosphatase increased activity, and urinary specific gravity and creatinine below reference values. On abdominal ultrasonography left adrenal gland measured 2.08 cm x 1.08 cm and the right adrenal gland measured 2.11 cm x 0.84 cm, indicating bilateral adrenomegaly compatible with PDH. In the hypogastric abdomen, a large heterogeneous hypoechogenic mass was also observed, with areas of cystic cavities, measuring 5.80 cm x 7.30 cm. A low dose dexamethasone suppression test (LDDST) was performed, due to PDH suspicion. The test did not confirm HAC, suspecting, then, to be a case of occult/atypical HAC. Due to the strong clinical suspicion, and owner financial problems for further investigated occult HAC, trilostane treatment was initiated.[...]

Parthenogenetic bovine embryos secrete type I interferon capable of stimulating ISG15 in luteal cell culture

Interferon tau (IFNT) is the pregnancy recognition signal in ruminants and is secreted by trophoblast cells. Paracrine action in the endometrium is well established by inhibiting luteolytic pulses of prostaglandin F2 alpha. Recently, endocrine action was documented in the corpus luteum, blood cell and liver. It was hypothesized that conditioned medium (CM) obtained from days 7, 9 and 12 parthenogenetic embryos alters luteal cell gene expression. The aim was to establish a bovine mixed luteal cell culture to evaluate cellular response associated to interferon stimulated genes, steroidogenesis and apoptosis. Conditioned medium was obtained from Days 7, 9 and 12 parthenogenetic (PA) embryos culture. Moreover, antiviral assay was performed on CM from Days 7, 9 and 12 to verify Type I interferon activity. Luteal cell culture was validated by steroidogenic and apoptotic genes (CYP11A1, HSD3B1, BAX, BCL2, AKT and XIAP mRNA expression), and concentration of progesterone as endpoint. Luteal cell culture was treated with interferon alpha (IFNA) and CM from parthenogenetic embryos. Antiviral assay revealed Type I interferon activity on CM from embryos increasing on Days 9 and 12. ISG15 mRNA was greater in the mixed luteal cells culture treated with 1, 10 and 100ng/ml of interferon alpha (IFNA) and also on Days 7, 9 and 12 CM treatments. Concentration of progesterone was not altered in luteal cell culture regardless of treatments. Steroidogenic and apoptotic genes were similar among groups in luteal cell culture treated with different doses of IFNA or CM from PA embryos. In conclusion, parthenogenetic embryo-derived CM has antiviral activity, luteal cell culture respond to Type I interferon by expressing IGS15. These data indicate this model can be used for IFNT endocrine signaling studies.(AU)