Avaliação da reserva ovariana na predição do mau prognóstico reprodutivo: quando e como investigar / Ovarian reserve evaluation in the prediction of poor reproductive prognosis: when and how to investigate?

Muitas vezes, torna-se um grande desafio para o ginecologista a identificação daquelas com maior ou menor chance de concepção. Vários marcadores laboratoriais e ultrassonográficos, conhecidos conjuntamente como testes de avaliação da reserva ovariana, são estudados há décadas com a intenção de se buscar uma ferramenta para a predição do potencial reprodutivo. E, embora ainda se busquem os marcadores ideais para aplicação clínica, mais difícil do que os definir é definir quando eles estão indicados. Este artigo de atualização, assinado pela Comissão Nacional Especializada em Ginecologia Endócrina da Febrasgo, pretende oferecer ao leitor as ferramentas necessárias para o uso racional dos testes de avaliação da reserva ovariana no cotidiano.(AU)

Often, it becomes a great challenge for the gynecologist to identify women with a greater or lesser chance of conception. Several laboratory and ultrasound markers, known jointly as ovarian reserve evaluation tests, have been studied for decades with the intention of seeking a tool for the prediction of reproductive potential. And, while the ideal markers for clinical application are still sought, defining them is as harder as defining when they are indicated. This update article, signed by the National Specialized Committee on Gynecologic Endocrinology, Febrasgo, intends to offer the reader the necessary tools for the rational use of ovarian reserve evaluation tests in daily practice.(AU)

Relação entre os níveis séricos do hormônio anti-Mulleriano, inibina B, estradiol e hormônio folículo estimulante no terceiro dia e o status folicular ovariano / Relationship of serum anti-Müllerian hormone, inhibin B, estradiol and FSH on day 3 with ovarian follicular status

OBJETIVO: examinar a hipótese de que o nível sérico do hormônio anti-Mülleriano (HAM) reflete o status folicular ovariano. MÉTODOS: Desenho: estudo prospectivo. Pacientes: foram incluídas 101 candidatas à FIV-TE submetidas à estimulação ovariana controlada com agonista de GnRH e FSH. Depois de atingir a supressão da hipófise e antes da administração de FSH (dia basal), os níveis séricos de HAM, inibina B e FSH foram avaliados. O número de folículos antrais foi determinado pela ultra-sonografia (dia basal) (folículo antral precoce; 3-10 mm). RESULTADOS: as médias do nível sérico de HAM, inhibina B, E2, P4 e FSH (dia basal) foram 3,4±0,14 ng/mL, 89±4,8 pg/mL, 34±2,7 pg/mL, 0,22±0,23 ng/mL e 6,6±0,1 mUI/mL, respectivamente, e a média do número de folículos antrais precoces foi 17±0,39. O nível sérico do HAM foi negativamente correlacionado com a idade (r= -0,19, p<0,04) e positivamente correlacionado com o número de folículos antrais precoces (r=0,65, p<0,0001), mas isto não se aplicou aos níveis séricos de inibina B, E2 e FSH. CONCLUSÕES: esse dado demonstra a associação do HAM com a quantidade de folículo antral, sendo aquele, portanto, um provável biomarcador do status folicular ovariano.

PURPOSE: to examine the hypothesis that serum anti-Müllerian hormone (AMH) levels reflect the ovarian follicular status. METHODS: Design: prospective study. Patients: we studied 101 IVF-ET candidates undergoing controlled ovarian hyperstimulation with GnRH agonist and FSH. After the achievement of pituitary suppression and before FSH administration (baseline), serum AMH, inhibin B, and FSH levels were measured. The number of antral follicles was determined by ultrasound at baseline (early antral follicles; 3-10 mm). RESULTS: at baseline, median serum levels of AMH, inhibin B, E2, P4 and FSH were 3.42±0.14 ng/mL, 89±4.8 pg/mL, 34±2.7 pg/mL, 0.22±0.23 ng/mL and 6.6±0.1 mIU/mL, respectively, and the mean number of early antral follicles was 17±0.39. Serum levels of AMH were negatively correlated with age (r=-0.19, p<0.04), and positively correlated with number of antral follicles (r=0.65, p<0.0001), but this did not apply to serum levels of either inhibin B, E2 or FSH. CONCLUSION: the data demonstrate an association between AMH and antral follicular counts. Therefore, AMH is probable a biomarker of ovarian follicular status.

Concentrações hormonais e desenvolvimento folicular de vacas leiteiras em hipertermia sazonal e aguda / Hormone concentration and follicular development in dairy cows under seasonal and acute hyperthermia

Avaliaram-se as concentrações hormonais e os parâmetros de desenvolvimento folicular de vacas leiteiras expostas ao calor sazonal e agudo. Dividiram-se os animais em quatro grupos: verão (n=5), outono (n=5), inverno com hipertermia aguda (grupo câmara climática, (CC), n=5) e inverno (n=9). Os animais foram abatidos no sétimo dia após a ovulação, e os parâmetros de desenvolvimento folicular avaliados. O líquido folicular do maior folículo foi aspirado e armazenado para posterior análise de hormônios esteróides e inibina. O número de células da granulosa vivas no verão e no outono foi 40 e 45 por cento respectivamente, menor que no inverno (P<0,05). A concentração de estradiol (E2) no inverno foi 62 por cento maior que no outono (P<0,05) e 34 por cento superior ao grupo verão (P<0,06). Houve um aumento na quantidade de androstenediona no verão em relação aos grupos inverno (P<0,08) e outono (P<0,05). A concentração de inibina foi maior no inverno do que no verão e CC (P<0,05). A exposição ao calor sazonal e agudo modificou os parâmetros de desenvolvimento do folículo e as concentrações hormonais no líquido folicular, podendo explicar em parte a queda nas taxas de concepção no verão.

The present study evaluated the seasonal and acute heat stress on follicular development and steroid and inhibin concentrations in follicular fluid, in bovine dominant follicle. Cows were distributed into four treatments: summer (n=5), autumn (n=5), animals heat stressed during the winter (n=5) and winter (n=9). On day 7 of the estrous cycle, animals were slaughtered and parameters related to follicle development were evaluated. The follicular fluid (FF) was aspirated and stored for further hormonal analysis. During the summer, the number of viable granulosa cells was 40 percent lower than during the winter, and there was a 45 percent decrease in this parameter during the autumn (P<0.05). In the winter, estradiol concentration was 62 percent higher than during the autumn (P<0.05) and 42 percent higher than during the summer (P<0.06). There was an increase in androstenedione concentration in summer group, when compared to winter (P<0.08) and autumn (P<0.05) groups. Inhibin concentration was higher in winter groups than summer and winter heat stressed groups (P<0.05). Seasonal and acute heat stress altered developmental parameters in dominant follicle and hormonal concentration in follicular fluid, those effects can partially explain the decrease in conception rates during summer.

Mucinous cystadenoma coexisting with stromal tumor with minor sex-cord elements of the ovary: a case report

Mucinous neoplasms occur rarely in association with cystic teratoma, Sertoli-Leydig cell tumor, granulosa cell tumor or carcinoid tumor. Several cases of an ovarian stromal tumor with minor sex-cord elements have been reported in the literatures. However, there has been no report about an ovarian mucinous neoplasm coexisting with a stromal tumor with sex-cord elements yet. We report a case of an ovarian neoplasm composed of both mucinous cystadenoma and stromal tumor with minor sex-cord elements in a 58-yr-old female. The ovary including the mass measured 5 cm in size. On section, it revealed an unilocular cyst (4.5 cm in diameter) filled with mucinous fluid. There was a round, yellow, solid nodule, 1.5 cm in diameter within the wall. Microscopically, the cyst was lined by a single layer of endocervical mucinous epithelium and the nodule was composed of spindle cells showing an intersecting and whorled arrangement. There were cell nests showing polygonal shape with abundant cytoplasm among the spindle cells. They showed immunoreactivity for inhibin and did not have any connection with the adjacent mucinous epithelium. Therefore, we interpret the mucinous cystadenoma as having arisen de novo.

La inhibina (Inh) A producida por células de granulosa humanas actuaría como factor autocrino-paracino regulando el mecanismo de luteinización folicular / Inh A produced by the human granulosa cells would act as autocrinous-paracrinous factor regulating the follicular luteinization mechanism

Las Inh gonadales ejercen un efecto local en la regulación de la foliculogénesis. En éste trabajo se midió la producción de inhibina popr células de granulosa humana en cultivo por un enzimoinmunoensayo específico para las formas A y B encontrándose mayores niveles de Inh A que B (2,89ñ0,88 vs 0,025ñ0,008 ng/1 por 10 cels. n:6, p<0.05). Se detrminó la concentración de inhibinas, Progesterona, Estradiol, FSH y hCG en fluido folicular (FF) obtenido el día de la aspiración ovocitaria en 23 pacientes sometidas a FIV. Se encontró que la concentración de Inh A fue mayor que la de Inh B (6,87ñ1,27 vs 1,24ñ0,23 ng/mg proteína). Se observó una correlación positiva entre la concentración intrafolicular de Inh A y Progesterona (r=0,47 p<0.05), Estradiol (r=0,67 p<0.05) y FSH (r=0,74 p<0.05). También se observó correlación positiva entre los niveles de E y FSH en FF (r=0.56 p<0.05). La Inh B en FF no correlacionó, con ninguno de los parámetros estudiados. Se ensayó el efecto in vitro de la Inh. A en cultivo de células de granulosa inmaduras de rata, que posee la ventaja de tener mayor capacidad proliferativa que las humanas obtenidas por FIV. Se estimuló con FSH (20 ng/ml), Estradiol (500 ng/ml) y TGFß (5 ng/ml) durante 48 hs. Se observó inhibición (p<0,05) de la proliferación medida por incorporación de timidina. (Control: 5543ñ146; Inh 4 ng/ml: 4767ñ174; Inh 40 ng/ml: 3480ñ117 cpm). Esto fue confirmado por MTS (Absorbancia=Control: 1,3ñ0,2; Inh 40 ng/ml: 0,5ñ0,02; p<0,05). Concluímos: a) las células de granulosa humanas provenientes de FIV producen más Inh A que B cultivo, ésta misma relación se observa en FF, b) el contenido de Inh A en FF se correlaciona con la concentración de FSH y esteroides foliculares y c) la Inh A inhibe la proliferación de células de granulosa en cultivo. Estos resultados sugieren que la Inh A actuaría como factor autócrino/parácrino regulando el mecanismo de selección y/o luteinización folicular

Inibina-A no soro materno no segundo trimestre gestacional: um novo marcador bioquímico de síndrome de Down / Second-trimester maternal serum inhibin-A: a new biochemical marker of Down's syndrome

As inibinas säo hormônios glicoproteícos que durante a gravidez, säo sintetizados, principalmente, pela placenta. A inibina-A é a principal forma dimérica circulante durante este período. Este hormônio vem sendo incluído no teste combinado de rastreamento da síndrome de Down por marcadores bioquímicos de soro materno, por aumentar a taxa de detecçäo. Nosso estudo abrangeu a dosagem de inibina-A em 160 amostras de soro materno utilizando-se um ensaio imunoenzimático do tipo sanduíche, de média sensibilidade. Verificamos que os níveis de inibina-A säo constantes ao longo do segundo trimestre gestacional, como já havia constatado por outros autores anteriormente. Encontramos uma correlaçäo positiva entre os níveis de inibina-A e os de Beta-hcg (p=0,0001). O peso materno e a inibina-A sérica mostraram uma correlaçäo negativa (p=0,0023). Näo foi observada correlaçäo estatisticamente signicativa dos valores de inibina-A com níveis de AFP e uE3. Nossos dados foram comparados com aqueles encontrados na literatura médica. Este estudo nos permitirá incluir mais este marcador em nosso esquema de rastreamento pré-natal de síndrome de Down

Patterns of inhibin and FSH localization in endometrium of baboon (Papio anubis) during menstrual cycle and early pregnancy.

Immunoreactive 10.5 KDa moiety of inhibin and hFSH was present in the baboon endometrium during menstrual cycle, early pregnancy and in castrated animals treated with steroid hormones, estrogen and/or progesterone. Endometrial differences during the menstrual cycle altered the intensity of immunostaining of inhibin and FSH. Maximum staining was observed in late luteal phase for both the hormones. In early pregnancy (35th day), the conceptus increased the staining for inhibin in the adjoining endometrial glands. Treatment of castrated animals with steroids for 14 days caused increased staining for inhibin. Maximum staining was observed when treated with estradiol or progesterone, whereas combination of estrogen and progesterone treatment decreased the staining reaction. In conclusion, both inhibin and FSH were localized in baboon endometrium and were under the influence of estrogen and progesterone.

Seminal inhibin--prospects for immunocontraception.

Passive immunization of adult rats, hamsters and marmosets with rabbit anti-seminal inhibin resulted in complete or partial block of fertility. The antiserum treatment presumably neutralized endogenous inhibin resulting in an unopposed rise in circulating FSH. This probably led to a refractoriness of the testes to FSH resulting in complete spermatogenic arrest. Nevertheless, there was no change in the mating behaviour of the animals. The antibodies also affected the epididymal spermatozoa by causing large scale agglutination.

Dual regulatory action of prostatic inhibin (10.7 kDa) on DNA synthesis.

Present studies deal with the role of inhibin in proliferation and growth. The effect of inhibin on incorporation of 3H-thymidine in prostatic DNA in vivo as well as by NRK-49F and Balb/c3T3 cell lines in vitro, was investigated. Also studied the immunocytochemical localization of inhibin in normally proliferating and differentiated tissues of human prostate and endometrium. The in vivo studies revealed a suppression of 3H-thymidine uptake both in ventral (33%) and dorsolateral (26%) lobes of rat prostate. Interestingly, the histology of inhibin treated rat prostate manifested amidst the epithelial lining, an appearance of apoptotic bodies which are considered to be indicative of cell death. Further, the immunocytochemical studies for localization of inhibin showed intense staining in the differentiated human prostate and endometrium as compared to the respective proliferative tissues. Is inhibin kept suppressed in these proliferating tissues, because it is antiproliferative? The present in vitro experiments demonstrated that, at low inhibin concentrations, the incorporation of 3H-thymidine is stimulated while at higher doses it is suppressed. Thus, it is clear that prostatic inhibin seems to have a concentration-dependent dual role in the regulation of DNA synthesis.

De novo biosynthesis and localization of inhibin in marmoset (Callithrix jacchus) and rat epididymis.

Using specific polyclonal antibodies generated against a 13 Kd human testicular inhibin, immunocytochemical localization was carried out in epididymis of intact and castrated marmoset monkey and rat epididymis. Inhibin was found to be present in the cytoplasm of epithelial cells of caput, corpus and cauda epididymis. The intensity of staining and pattern of distribution did not change following castration. Further, the in vitro biosynthesis of inhibin studied by incorporating 3H-leucine and precipitating it with specific antibody indicated maximum biosynthesis in the corpus epididymis in case of marmosets and cauda in case of rats. Following castration in rats, the epididymal tissue still retained the capacity to biosynthesize inhibin. These studies indicate that marmoset and rat epididymis are capable of biosynthesizing/absorbing inhibin and whose synthesis does not depend on androgens.

Localization of inhibin in testes of human, bonnet monkey, dog and rat by immunoperoxidase technique.

Immunocytochemical study on the localization of inhibin in the testes of human, bonnet monkey, dog and rat was carried out using indirect immunoperoxidase technique, in order to investigate the cell types involved in inhibin production/storage. A positive reaction was observed in the testes of human, monkey and dog while it was negative in rat testis using specific antiserum to human testicular inhibin generated against homogeneous preparation of human testicular inhibin in our laboratory. Inhibin was found to be localized in Sertoli cells, spermatogonia and primary spermatocytes of human, monkey and dog testes. A weak positive reaction was observed in spermatids of human testis only. Interestingly, Leydig cells of human, monkey and dog testes showed positive reaction indicating presence of inhibin in these cells also.