Grado de desarrollo del folículo preovulatorio y su relación con el tamaño del cuerpo lúteo y la producción de progesterona en las razas Romosinuano (RS) y Brahman (Br) / Degree of development of the preovulatory follicle and its relationship with corpus luteum size and progesterone production in the Romosinuano (RS) and Brahman (Br) breeds

RESUMEN Para comparar en los folículos preovulatorios de hembras bovinas de las razas Brahman (Br) y Romosinuano (RS) el desarrollo de los diferentes componentes foliculares, como el tamaño del antro folicular, el grosor y el volumen ocupado por la capa de células de la granulosa (CG) y de la teca interna (TI), y su posible relación con el desarrollo del cuerpo lúteo (CL) y la producción de progesterona P4, se utilizaron 5 hembras de la raza RS y 5 de la raza Br. A estas se les realizó seguimiento ecográfico durante 2 ciclos estrales y se les efectuó muestreo de suero sanguíneo para determinar por Elisa los niveles de P4. En el tercer estro se les extirpó el ovario que contenía el folículo preovulatorio y se realizó un corte diametral sobre el estigma del folículo para seccionarlo en 2 partes iguales. En una de las partes se hicieron cortes histológicos y se determinó la morfometría folicular. La duración del ciclo estral en las hembras de la raza Br osciló entre 18 y 21 días, con promedio de 19,9 ± 1,6 días, y en las vacas RS entre 18 y 24 días, con un promedio 21,2 ± 1,69 días. La duración del estro fue de 13,9 ± 6,98 y 9,60 ± 4,72 h para las razas RS y Br, respectivamente. El intervalo estro-ovulación fue de 21,20 ± 5,07 h para los animales de la raza Br y de 24,40 ± 6,43 para los RS. No se registraron diferencias entre razas ni en el grosor (RS: 55,12 ± 6,46 μm vs. Br: 49,48 ± 17,07 μm), p > 0,05, tampoco en el volumen ocupado por la capa de CG (RS: 27,93 ± 6,11 mm1 vs. Br: 25,40 ± 13,85 mm3) de los folículos preovulatorios p < 0,05, en el grosor (RS: 122,50 ± 20,53 μm vs. Br: 129,61 ± 84,85 μm) o en el volumen (RS: 64,97 ± 19,71 mm1 vs. Br: 59,83 ± 25,67 mm3) de las células de la TI. El máximo desarrollo de los CL para la totalidad de las hembras se alcanzó el día 12,6 ± 4,05 (día 0 = estro), con un diámetro promedio de 22,92 ± 3,60 mm. Para las de la raza RS el diámetro máximo fue 23,06 ± 3,9 mm y se observó en promedio el día 14,88 ± 3,4. Para las de la raza Br fue el día 10,00 ± 3,16 con un promedio de 22,75 ± 3,16 mm. En ese día, la concentración media de P4 fue de 5,37 ± 1,38 ng/ml para la raza RS y 5,74 ± 0,89 ng/ml para la raza Br. No se presentaron diferencias significativas entre razas (p > 0,05). Se concluyó que no existen diferencias en los eventos fisiológicos estudiados entre las razas RS y Br. Los hallazgos del presente estudio, sin duda, pueden servir de base para futuros análisis en los bovinos residentes en el trópico.

ABSTRACT To compare in the preovulatory follicles of bovine females of the Brahman (Br) and Romosinuano (RS) breeds the development of the different follicular components, such as the size of the follicular antrum, the thickness and volume occupied by the granulosa (CG) and theca interna cells (TI) layers and their possible relationships with the development of the corpus luteum (CL) and progesterone (P4) production, 5 RS and 5 Br females were used; ultrasound follow-up was performed during 2 estrous cycles and blood serum sampling was carried out to determine progesterone (P4) levels by Elisa, at the third estrus, the ovary containing the preovulatory follicle was removed, and a diametral cut was made on the stigma of the follicle to divide it into 2 equal parts. Histological sections were made of one of the parts and follicular morphometry was determined. Duration of the estrous cycle in Br females ranged between 18 and 21 days, with an average of 19,9 ± 1,6 days and between 18 and 24 with an average of 21,2 ± 1,69 days for RS cows. Estrus duration was 13,9 ± 6,98 and 9,60 ± 4,72 h for RS and Br breeds, respectively. The estrus-ovulation interval was 21,20 ± 5,07 h for the Br breed animals and 24,40 ± 6,43 RS. There were no differences between breeds or in thickness (RS: 55,12 ± 6,46 μm vs. Br: 49,48 ± 17,07 μm) p > 0,05, nor in the volume occupied by the granulosa cells layer (RS: 27,93 ± 6,11 mm3 vs. Br: 25,40 ± 13,85 mm3) of pre-ovulatory follicles; neither in the thickness (RS: 122,50 ± 20,53 vs. Br: 129,61 ± 84,85 μm) nor in the volume (RS: 64,97 ± 19,71 mm3 vs. Br: 59,83 ± 25,67 mm3) of the internal theca cells; p < 0,05. The maximum CL development for all the females was observed on day 12,6 ± 4,05 (day 0 = estrus), with an average diameter of 22,92 ± 3,60 mm. For those of the RS breed the maximum diameter was observed on average on day 14,88 ± 3,4, with 23,06 ± 3,9 mm, while for those of the Br breed it was day 10,00 ± 3,16 with an average diameter of 22,75 ± 3,16 mm; at this time the mean progesterone concentration was 5,37 ± 1,38 ng/ml for the RS breed and 5,74 ± 0,89 ng/ml for the Br breed. There were no significant differences between breeds (p-value: 0,5561). It was concluded that there are no differences in the physiological events studied between the studied breeds. Present results, undoubtedly, might serve as basis to future studies in bovines in tropical zones.

Tumores Benignos de Ovario en la edad Pediátrica y Adolescencia / Benign Ovarian Tumors in Pediatric and Adolescence

Introducción: Los tumores ováricos son raros en la edad pediátrica y representan del 1 al 5% de los tumores infantiles. El objetivo del presente trabajo es conocer cuáles son los tumores de ovario más frecuentes en nuestro medio en niñas y adolescentes en un centro oncológico de la ciudad de Guayaquil-Ecuador. Métodos: El presente estudio es descriptivo y retrospectivo, en la que se realiza una revisión de registros de niñas y adolescentes con diagnóstico de tumores de ovario, en el Instituto Oncológico Nacional "Dr. Juan Tanca Marengo" en el período de enero 2015 a mayo del 2017. Las variables registradas fueron edad, diagnóstico ecográfico, diagnostico histopatológico, ovario afecto, características clínicas, salpingectomía, marcadores tumorales. Se utiliza estadística descriptiva. Resultados: Ingresaron al estudio 32 pacientes de 12.7 años promedio. El ovario más afectado fue el derecho con una relación 2.2: 1 y en 9 % fue bilateral. La histología reportada de los quistes serosos fueron el 21 % quiste folicular y 18.5 % por quiste lúteo. Los quistes simples representaron el 15.25 %, el cistoadenoma y el teratoma maduro con un 9 %, el quiste dermoide y le endometrioma con un 3 %. El dolor fue el síntoma más llamativo estado como síntoma único en 65.5 %. El 18.75 % terminó en anexectomía con ooforectomía por extensión del quiste, el 81.25 % solo requirió quistectomía. Conclusión: La histología reportada en esta serie de tumores benignos de ovario, los quistes serosos foliculares fueron los más prevalentes, seguidos de los quistes lúteos. El dolor pélvico fue el síntoma más llamativo. La mayoría de los casos requirieron quistectomía y el 19 % requirió adicionalmente ooforectomía.

Introduction: Ovarian tumors are rare in pediatric age and represent 1 to 5% of childhood tumors. The objective of this work is to know which are the most frequent ovarian tumors in our environment in girls and adolescents in an oncological center of the city of Guayaquil-Ecuador. Methods: The present study is descriptive and retrospective, in which a review of records of girls and adolescents diagnosed with ovarian tumors is performed, at the National Oncology Institute "Dr. Juan Tanca Marengo" in the period from January 2015 to May 2017. The variables recorded were age, ultrasound diagnosis, histopathological diagnosis, affected ovary, clinical characteristics, salpingectomy, tumor markers. Descriptive statistics are used. Results: 32 patients with an average age of 12.7 entered the study. The most affected ovary was the right one with a 2.2: 1 ratio and in 9% it was bilateral. The reported histology of serous cysts were 21% follicular cyst and 18.5% by luteal cyst. Simple cysts accounted for 15.25%, cystadenoma and mature teratoma with 9%, dermoid cyst and endometrioma with 3%. Pain was the most striking condition as a single symptom in 65.5%. 18.75% ended up in anexectomy with oophorectomy due to cyst extension, 81.25% only required a cystectomy. Conclusion: The histology reported in this series of benign ovarian tumors, follicular serous cysts were the most prevalent, followed by luteal cysts. Pelvic pain was the most striking symptom. The majority of cases required a cystectomy and 19% additionally required oophorectomy.

Melatonin effects on ovarian follicular cells: a systematic review

SUMMARY Melatonin is known for its effects on both the sleep and reproductive system of mammals. The latter has melatonin receptors type 1 and 2, which act to regulate, among other things, cyclic AMP. Notwithstanding all the literature data, there is still no sound knowledge or a clear understanding of the hormone's action on the physiology of ovarian follicular cells. OBJECTIVE To review and evaluate studies about melatonin action on the ovarian granulosa/theca interna cells from the literature. METHODS The systematic review was carried out according to the PRISMA recommendations. The MEDLINE and Cochrane primary databases were consulted with the use of specific terms. There was no limitation on language or publication year. RESULTS Seven papers about melatonin action on granulosa cells were selected. The following can be attributed to the hormone's effects: a) progesterone increase in culture medium; b) increased estrogen production; c) antagonistic action on estrogen; d) improvement in cell quality resulting in improved embryo and higher pregnancy rates; e) improved cell proliferation via MAPK; f) reduction of free radicals. Nevertheless, there are contrarian papers reporting a reduction in progesterone production. CONCLUSION Melatonin interferes in sex steroid production, boosting progesterone output. Such action may help improve oocyte quality.

RESUMO A melatonina é conhecida por seus efeitos no sono e no sistema reprodutivo dos mamíferos. Este último tem receptores de melatonina tipos 1 e 2, que atuam para regular, entre outras coisas, o AMP cíclico. Apesar de todos os dados da literatura, ainda não há um conhecimento sólido ou uma compreensão clara da ação do hormônio na fisiologia das células foliculares ovarianas. OBJETIVO Revisar e avaliar estudos da ação da melatonina na literatura sobre as células internas da granulosa/teca ovariana. MÉTODOS A revisão sistemática foi realizada de acordo com as recomendações do Prisma. As bases de dados primárias Medline e Cochrane foram consultadas com o uso de termos específicos. Não houve bar na língua ou ano de publicação. RESULTADOS Sete artigos sobre a ação da melatonina nas células da granulosa foram selecionados. O que se segue pode ser atribuído aos efeitos do hormônio: a) aumento de progesterona no meio de cultura; b) aumento da produção de estrogênio; c) ação antagônica no estrogênio; d) melhoria na qualidade celular, resultando em melhor embrião e maiores taxas de gravidez; e) melhor proliferação celular via MAPK; f) redução de radicais livres. No entanto, existem artigos controversos relatando redução na produção de progesterona. CONCLUSÃO A melatonina interfere na produção de esteroides sexuais, aumentando a produção de progesterona. Tal ação pode ajudar a melhorar a qualidade do oócito.

Melatonin may prevent or reverse polycystic ovary syndrome in rats

SUMMARY OBJECTIVE To evaluate the ovarian effects of melatonin (Mel) in a rat model of polycystic-ovary-syndrome (PCOS) before and after permanent estrus induction. METHODS Thirty-two adult-female rats with regular estrous cycle were equally divided into four groups: 1) GCtrl - at estrous phase. 2) GPCOS - at permanent-estrous phase. 3) GMel1 - treated for 60 days with Mel (0.4 mg/Kg) during permanent estrus induction and 4) GMel2 - rats with PCOS and treated for 60 days with Mel. After that, the animals were euthanized, and the ovaries were removed and processed for paraffin embedding. Sections were stained with H.E. for histomorphometry or subjected to immunohistochemistry for Ki-67 and cleaved caspase-3 (Casp-3) detections. RESULTS The GPCOS showed lack of corpus luteum and several ovarian cysts, as well as interstitial-like cells. The presence of corpus luteum and a significant increase in primary and antral follicles were observed in Mel-treated groups, which also showed a decrease in the number of ovarian cysts and in the area occupied by interstitial-like cells. These results were more evident in GMel1. The percentage of Ki-67-positive cells was significantly higher in the Mel-treated groups, mainly in the GMel2, as compared to GPCOS. On the other hand, the percentage of Casp-3-positive cells was significantly lower in granulosa cells of GMel1, whereas it was significantly higher in the interstitial-like cells of GMel2, in comparison to GPCOS. CONCLUSION Melatonin administration prevents the permanent estrus state in the PCOS rat model. This effect is more efficient when melatonin is administered before permanent estrus induction.

RESUMO OBJETIVO Avaliar os efeitos ovarianos da melatonina (Mel) em ratas com síndrome dos ovários policísticos (SOP) antes e após a indução do estro-permanente. MÉTODOS Trinta e duas ratas com ciclos estrais regulares foram igualmente divididas em quatro grupos: 1) GCtrl - fase de estro. 2) GSOP - fase de estro-permanente. 3) GMel1 - tratadas por 60 dias com Mel (0,4 mg/kg) durante a indução do estro-permanente e 4) GMel2 - ratas com SOP e tratadas com Mel. Após eutanásia dos animais, os ovários foram processados para inclusão em parafina. Cortes foram corados com H.E ou submetidos à imuno-histoquímica para detecção de Ki-67 e caspase-3 clivada (Casp-3). RESULTADOS O GSOP mostrou ausência de corpos lúteos e vários cistos ovarianos, além de inúmeras células intersticiais. A presença de corpos lúteos e o aumento significativo dos folículos primários e antrais foram observados nos grupos tratados com Mel, os quais também mostraram diminuição no número de cistos ovarianos e na área ocupada pelas células intersticiais. Esses resultados foram mais evidentes no GMel1 do que no GMel2. A porcentagem de células Ki-67 positivas foi significativamente maior no GMel1 e no GMel2, sendo mais evidente no GMel2, em comparação ao GSOP. Por outro lado, a porcentagem de células positivas à Casp-3 foi menor nas células da granulosa do GMel1 e maior nas células intersticiais do GMel2, em comparação ao GSOP. CONCLUSÃO A administração de melatonina previne o estado de estro-permanente em ratas com SOP. Esse efeito é mais eficiente quando a melatonina é administrada após indução do estado de estro-permanente.

Energy Demand and Its Regulatory Mechanism during Folliculogenesis / 中国医学科学院学报

The growth and development of follicles are regulated by genes,hormones and growth factors autocrined and paracrined from granulosa cells,theca cells,and oocytes.Products of glycolysis from granulosa cells such as pyruvate and lactate are one of the main energy sources,which play an important role during folliculogenesis and follicle maturity.Studies on the changes of the products and rate-limiting enzymes during granulosa cells' glycolysis help to clarify the molecular mechanism of energy demand in folliculogenesis and guide the clinical treatment of infertility due to abnormal follicular development.This article reviews recent research advances in the energy demand and regulatory mechanism in different states of folliculogenesis.

Adult granulosa cell tumors of the ovary: a retrospective study of 30 cases with respect to the expression of steroid synthesis enzymes / 부인종양

OBJECTIVE: Some, but not all, granulosa cell tumors are characterized by estrogen production. This study was designed to determine whether there are clinical or pathological variations in granulosa cell tumors in relation to the expression of sex steroid synthesis enzymes. METHODS: Clinical symptoms, serum hormonal values, and histology of 30 granulosa cell tumor patients who underwent surgery between 2002 and 2014 were retrospectively reviewed. RESULTS: Most patients presented with abnormal genital bleeding including abnormal menstrual cycles. Eight of 16 patients older than 50 years had endometrial hyperplasia and one had endometrial cancer. Serum 17β-estradiol (E2) levels tended to be higher in patients over 50 years of age (p=0.081). Serum follicle-stimulating hormone (FSH) levels were low in all patients irrespective of serum E2 levels. Magnetic resonance imaging revealed a thicker endometrium in older as compared to younger patients (p<0.05). Tumor cells in the majority of cases were positive for inhibin α and P450 aromatase, irrespective of age and serum E2 levels. P450 17α-hydroxylase (P450c17) expression varied among cases. P450c17 was strongly positive in luteinized tumor cells and weakly positive in theca cells and fibroblasts. High E2 levels were associated with P450c17-positive cells in the tumor (p<0.05). CONCLUSION: The expression of hormone-synthesizing enzymes divides granulosa cell tumors into 2 distinct types; tumors with P450c17-positive cells show elevated serum E2 and related clinical symptoms, while tumors without these cells show symptoms related to FSH suppression by inhibin.

Unusual malignant neoplasms of ovary in children: two cases report / 소아과

Sex cord tumors with annular tubules are known to originate from the sex cord of embryonic gonads that synthesize Sertoli cells, Leydig cells, granulosa cells, and theca cells of the ovarian stroma, while ovarian small cell carcinoma of the hypercalcemic type is a type of neuroendocrine tumor. Both these tumors are uncommon, potentially malignant neoplasms in children. We report the case of a sex cord tumor with annular tubules in an 11-year-old girl and a case of small cell carcinoma of the hypercalcemic type in a 10-year-old girl. We also discuss the prognosis and management of these tumors.

Influencia de la 6-Bencilaminopurina y el agente gelificante en la reducción de la hiperhidricidad en Tectona grandis L / Influence of 6-Benzyladenine and gelling agent on the reduction of hyperhydricity in Tectona grandis L

The influence of different factors on shoot proliferation and the occurrence of hyperhydricity in teak (Tectona grandis L.) have been studied. Four concentrations of BA (2.22, 4.44, 6.66 and 8.88 µM) and a control treatment with 0 BA were examined. Aiming at reducing the costs during commercial propagation by using gelrite in stead of agar, the use of both gelling agent in the proliferation and hyperhydricity was tested. In order to evaluate if hyperhydricity can be reduced by increasing the gelrite concentration in the culture medium, three concentrations (2.0, 2.5 and 3.0 g L-1) were tested in combination with 4.44 µM BA. The proliferation and occurrence of hyperhydricity during 21 successive subcultures were evaluated. The highest proliferation was achieved in the treatments with 6.66 or 8.88 µM BA. They yielded 5.22 and 5.56 shoots/explant, respectively. But also, the highest percent of hyperhydric shoots was achieved in this treatment. Gelrite resulted in a higher proliferation, but also an almost two times higher hyperhydricity as compared to agar-solidified media. Satisfactory reduction in hyperhydricity (18%) was achieved with 3.0 g L-1 gelrite. However, the successive subcultures onto proliferation in this treatment favored hyperhydricity compromising shoot quality and it´s competence to proliferate. in vitro teak plants were ex vitro rooted and then transferred to greenhouse conditions for acclimatization; ten weeks after transfer they were ready for field plantation.

Se estudió la influencia de diferentes factores en la proliferación y la ocurrencia de la hiperhidricidad in teca (Tectona grandis L.). Se probaron cuatro concentraciones de BA (2,22; 4,44; 6,66 and 8,88 µM) y un control sin BA. Con el objetivo de reducir los costos durante la propagación comercial se experimentó sustituir el agar por el gelrite, para lo cual se estudió en efecto de ambos gelificantes en la proliferación y la hiperhidricidad de los brotes. Se estudiaron, tres concentraciones de gelrite (2,0; 2,5 and 3,0 g L-1) combinadas con 4,44 µM BA, con el objetivo de evaluar si la hiperhidricidad podía ser reducida incrementando la concentración de gelrite. Se evaluó la proliferación de brotes y la ocurrencia de la hiperhidricidad durante 21 subcultivos. Se logró una alta proliferación de brotes en los tratamientos con 6,66 y 8,88 µM BA (5,22 y 5,56 brotes), pero el porcentaje de brotes hiperhídricos también se incrementó. El gelrite resultó en una alta proliferación de brotes, pero con mayor incidencia de la hiperhidricidad que el medio gelificado con agar. Se obtuvo una reducción satisfactoria de la hiperhidricidad (18%), cuando la concentración de gelrite se incrementó hasta 3,0 g L-1. No obstante, la multiplicación de los brotes en este tratamiento más allá del 11no subcultivo favoreció la hiperhidricidad, lo que afectó la calidad de los brotes y su competencia para la proliferación. Las plantas fueron enraizadas ex vitro, transferidas a condiciones de invernadero para su aclimatización y diez semanas después de la transferencia estaban listas para la plantación en campo.

Morfologia das células intersticiais de ovários policísticos de ratas: um estudo experimental / Morphology of the interstitial cells of rat polycystic ovaries: an experimental study

OBJETIVOS: Avaliar a histomorfometria das células intersticiais dos ovários, bem como analisar a concentração sanguínea de esteroides sexuais de ratas portadoras de ovários policísticos induzidos pela luz contínua. MÉTODOS: Vinte ratas foram divididas em dois grupos: ratas na fase de estro (GCtrl ) e ratas portadoras de ovários policísticos induzidos pela iluminação contínua (GOP). Os animais do GCtrl permaneceram com período de luz das 7:00 s 19:00 horas, e os animais do GOP, com iluminação contínua (400 Lux), durante um período de 60 dias. Ao final desse período todos os animais foram anestesiados, foi coletado o sangue, para determinação dos níveis séricos de estradiol (E2), progesterona (P4) e testosterona (T), seguido da retirada dos ovários que foram fixados em formol a 10% e processados para inclusão em parafina. Cortes histológicos com 5 µm corados pela hematoxilina e eosina foram utilizados para análise histomorfométrica. As análises morfológicas, contagem de cistos, determinação da concentração e do volume nuclear das células intersticiais foram realizadas com o auxílio de microscópio de luz adaptado a uma câmera de alta resolução (AxioCam), cujas imagens foram transmitidas e analisadas em computador com software AxioVision Rel 4.8 (Carl Zeiss). Os dados obtidos foram submetidos ao teste t de Student (p<0,05). RESULTADOS: A morfologia mostrou a presença de cistos nos ovários pertencentes ao Grupo OP e de corpos lúteos no GCtrl, mostrando ainda evidências da origem das células intersticiais a partir das células da teca interna desses cistos. Com relação aos níveis hormonais o GOP apresentou níveis séricos de estradiol (pg/mL) aumentados em relação ao GCtrl (GOP=124,9±4,2>GCtrl=73,2±6,5; p<0,05), o mesmo ocorrendo com os níveis de testosterona (pg/mL) (GOP=116,9±4,6>GCtrl=80,6±3,9; p<0,05). Entretanto os níveis de progesterona (ng/mL) foram mais elevados no GCtrl em relação ao GOP (GCtrl=16,3±2,0>GOP=4,2±1,5; p<0,05). A morfometria mostrou haver aumento significante do volume nuclear no grupo GOP (GOP=102,1±5,2>GCtrl=63,6±16,5; p<0,05), assim como da área ocupada (%) pelas células intersticiais (GOP=24,4±6,9>GCtrl=6,9±3,2; p<0,05) em relação aos animais do GCtrl. CONCLUSÃO: As células intersticiais do ovário policístico da rata provavelmente provêm dos cistos ovarianos devido degeneração das células da granulosa e diferenciação das células da teca interna. As elevações dos níveis séricos de testosterona e de estradiol provavelmente provêm do aumento significativo da atividade celular e da área ocupada pelas células intersticiais.

PURPOSES: To evaluate the histomorphometry of ovarian interstitial cells, as well as the blood sex steroid concentrations of female rats with polycystic ovaries induced by continuous light. METHODS: Twenty female rats were divided into two groups: Control Group - in the estrous phase (CtrlG), and a group of rats with polycystic ovaries induced by continuous illumination (POG). CtrlG animals were maintained on a light period from 07:00 a.m. to 07:00 p.m., and POG animals with continuous illumination (400 Lux) for 60 days. After this period all animals were anesthetized and blood was collected for the determination of serum estradiol (E2), progesterone (P4), and testosterone (T), followed by removal of the ovaries that were fixed in 10% formalin and processed for paraffin embedding. Five-µm histological sections were stained with hematoxylin and eosin and used for histomorphometric analysis. Morphological analyses, cyst count, determination of concentration and of the nuclear volume of interstitial cells were performed with the aid of a light microscope adapted to a high resolution camera (AxioCam), whose images were transmitted to and analyzed by the computer using AxioVision Rel 4.8 software (Carl Zeiss). Data were analyzed statistically by the Student's t-test (p<0.05). RESULTS: Morphological analysis showed the presence of ovarian cysts in POG animals and corpora lutea in CtrlG animals, as well as evidence of the origin of interstitial cells from the internal theca of these cysts. POG animals presented increased serum estradiol levels (pg/mL) compared to CtrlG animals (POG=124.9±4.2>CtrlG=73.2±6.5, p<0.05), the same occurring with testosterone levels (pg/mL) (POG=116.9±4.6>CtrlG=80.6±3.9, p<0.05). However, progesterone levels (ng/mL) were higher in CtrlG than in POG animals (CtrlG=16.3±2.0>POG=4.2±1.5, p<0.05). Morphometry showed a significant increase in nuclear volume in POG animals (POG=102.1±5.2>CtrlG=63.6±16.5, p<0.05), as well as in the area occupied (%) by interstitial cells (POG=24.4±6.9>CtrlG=6.9±3.2, p<0.05) compared to CtrlG animals. CONCLUSION: The interstitial cells of the rat polycystic ovary probably originate from ovarian cysts due to the degeneration of granulosa cells and differentiation of the internal theca cells. The elevations of serum testosterone and estradiol were probably due to the significant increase in cell activity and in the area occupied by interstitial cells.

Regulation and 3 dimensional culture of tertiary follicle growth / 대한생식의학회지

It has been revealed that multiple cohorts of tertiary follicles develop during some animal estrous cycle and the human menstrual cycle. To reach developmental competence, oocytes need the support of somatic cells. During embryogenesis, the primordial germ cells appear, travel to the gonadal rudiments, and form follicles. The female germ cells develop within the somatic cells of the ovary, granulosa cells, and theca cells. How the oocyte and follicle cells support each other has been seriously studied. The latest technologies in genes and proteins and genetic engineering have allowed us to collect a great deal of information about folliculogenesis. For example, a few web pages (http://www.ncbi.nlm.nih.gov; http://mrg.genetics.washington.edu) provide access to databases of genomes, sequences of transcriptomes, and various tools for analyzing and discovering genes important in ovarian development. Formation of the antrum (tertiary follicle) is the final phase of folliculogenesis and the transition from intraovarian to extraovian regulation. This final step coordinates with the hypothalamic-pituitary-ovarian axis. On the other hand, currently, follicle physiology is under intense investigation, as little is known about how to overcome women's ovarian problems or how to develop competent oocytes from in vitro follicle culture or transplantation. In this review, some of the known roles of hormones and some of the genes involved in tertiary follicle growth and the general characteristics of tertiary follicles are summarized. In addition, in vitro culture of tertiary follicles is also discussed as a study model and an assisted reproductive technology model.

An Autopsy Case of Severe Ovarian Hyperstimulation Syndrome with Multifocal Arterial and Venous Thromboembolism / 대한법의학회지

Ovarian hyperstimulation syndrome (OHSS) is a rare, serious complication of assisted reproductive procedures for fertility treatment. Although most cases of OHSS are mild to moderate in nature, its severe form causes life-threatening complications including respiratory distress, renal failure, liver dysfunction, and thromboembolism because of hemoconcentration. We herein report a rare autopsy case of severe OHSS in a 29-year-old woman who underwent controlled ovulation induction. Three days after the administration of human chorionic gonadotropin, she developed abdominal pain and distension. Two days after, she suddenly developed respiratory arrest, hypotension and semicomatose mentality and died of respiratory failure. At autopsy, multifocal thromboembolism was detected in the internal carotid arteries, internal jugular veins, pulmonary arteries, inferior vena cava, renal arteries, and deep leg veins. And there were cerebral edema, anasarca, pleural effusion, massive pulmonary edema, ascites, and bilateral ovarian enlargement. Microscopically, the bilateral ovaries showed multiple well-developed cyst formations consisting of membrana granulosa and theca interna. To the best of our knowledge, this is the first autopsy report of severe OHSS with multifocal arterial and venous thromboembolism.

Potencial fisiológico de diásporos de Tectona grandis L.f. derrubados pelo vento / Physiological potential of diasporas of Tectona grandis L.f. felled by wind

Nas áreas de produção de sementes de Tectona grandis em Cáceres, MT, ventos fortes podem causar queda precoce de frutos. Diásporos de três lotes de teca, um obtido após a queda natural e os outros dois após queda precoce, foram inicialmente qualificados quanto ao teor de água, massa de mil diásporos, número de diásporos por quilograma, emergência em canteiro e respectivo tempo médio. Essas duas últimas variáveis foram novamente determinadas em intervalos de 50 dias até 331 dias após a colheita com o uso de 200 diásporos divididos em quatro repetições. Para os diásporos de queda natural, a emergência de plântulas e o tempo médio não diferiram nos períodos de análise com média geral de 60,6% e 10,8 dias. Os diásporos dos lotes de colheita precoce apresentaram a emergência inicial de 23,5 e 17,5% e os tempos médios de 19,4 e 22,6 dias, respectivamente. Até 115 dias após a colheita, a emergência aumentou (55,5 e 59,5%) e o tempo médio diminuiu (15,0 e 14,1 dias) e a partir daí ocorreu o inverso. Os diásporos de colheita precoce derrubados pelo vento apresentam maturação e o maior potencial fisiológico aos 115 dias após a colheita. No período de 64 a 269 dias após a colheita, esses diásporos atingem o padrão mínimo estabelecido para a teca e o período entre 115 a 165 dias é o mais adequado para sua utilização.

In the seed production areas of Tectona grandis in Cáceres, state of Mato Grosso, Brazil, strong winds cause the precocious fall of fruits. Diasporas of three lots of teak, one of then crop after the natural fall, and the others two after the precocious fall were characterized initially by the content of water, thousand diasporas mass, number of diasporas by kilogram, emergency in seedbeds and respective average time. With 200 diasporas divided in four replications and in periods of 50 days until 331 days after crop, the last two variables were determined again. To the diasporas of natural fall, the seedlings emergency and the average time were not different in periods of analyses with a general average of 60.6% and 10.8 days. The diasporas of the precocious crop lots presented respectively the initial emergency of 23.5 and 17.5% and the average time of 19.4 and 22.6 days. Until 115 days after crop, the emergency increased (55.5 and 59.5%) and the average time diminished (15.0 and 14.1 days) and on this point the inverse occurred. The diasporas of precocious crop fall by the wind present maturation and the greater physiological potential at 115 days after the crop. In the period at 64 to 269 days after crop, these diasporas reaches the established minimum standard for the teak and at 115 to 165 days is more adjusted period for its use.

Identification of Differentially Expressed Genes in Bovine Follicular Cystic Ovaries

Follicular cystic ovary (FCO) is one of the most frequently diagnosed ovarian diseases and is a major cause of reproductive failure in mammalian species. However, the mechanism by which FCO is induced remains unclear. Genetic alterations which affect the functioning of many kinds of cells and/or tissues could be present in cystic ovaries. In this study, we performed a comparison analysis of gene expression in order to identify new molecules useful in discrimination of bovine FCO with follicular cystic follicles (FCFs). Normal follicles and FCFs were classified based on their sizes (5 to 10 mm and > or =25 mm). These follicles had granulosa cell layer and theca interna and the hormone 17beta-estradiol (E2)/ progesterone (P4) ratio in follicles was greater than one. Perifollicular regions including follicles were used for the preparation of RNA or protein. Differentially expressed genes (DEG) that showed greater than a 2-fold change in expression were screened by the annealing control primer (ACP)-based PCR method using GeneFishing(TM) DEG kits in bovine normal follicles and FCFs. We identified two DEGs in the FCFs: ribosomal protein L15 (RPL15) and microtubule-associated protein 1B (MAP1B) based on BLAST searches of the NCBI GenBank. Consistent with the ACP analysis, semi-quantitative PCR data and Western blot analyses revealed an up-regulation of RPL15 and a down-regulation of MAP1B in FCFs. These results suggest that RPL15 and MAP1B may be involved in the regulation of pathological processes in bovine FCOs and may help to establish a bovine gene data-base for the discrimination of FCOs from normal ovaries.

Hyperreactio luteinalis associated with pregnancy induced hypertension

This is a unique case of hyperreactio luteinalis associated with partial HELLP syndrome, chronic superimposed hypertension, and intrauterine growth restriction. The cysts were managed conservatively. A caesarean section was performed at 34 weeks due to the deterioration of liver function tests, and intrauterine growth restriction. Multiple tissue biopsies of the ovaries were taken which showed theca leutein cysts. Her postoperative recovery was uneventful. The size of the cysts subsequently regressed to normal in 3 months postpartum

Pathologic Characteristics of Ovarian Hemorrhagic Polycyst in Estrogen Receptor-alpha (ERalpha) Knockout Mice and Roles of ERalpha in Hemorrhagic Polycyst

BACKGROUND: Polycystic ovary syndrome (PCOS) is the most common endocrinopathy causing anovulation in women of childbearing age. It has been well established that estrogen receptor-alpha knockout (ERalphaKO) mice display several pathologic ovarian phenotypes of PCOS. The aims of this study were to determine ovarian pathology in new ERalphaKO mice using a CreloxP approach and intra-ovarian ERalpha function as regulating key aspects of PCOS. METHODS: ERalphaKO mice, which were deficient in exon 3 of the ERalpha gene, were used. Immunohistochemical studies were done on ovaries of control and ERalphaKO mice using antibodies specific to ERalpha, ERbeta, inhibin-alpha, and alpha-smooth muscle actin (SMA), as well as histochemical staining using Sudan black-B. RESULTS: All ovaries of ERalphaKO mice were larger than control mouse ovaries and displayed a disrupted theca-interstitial tissue organization, multiple atretic follicles and multiple hemorrhagic cysts. None of the ERalphaKO mouse ovaries showed a corpus luteum. In addition, heavy deposition of Sudan black-B positive foamy cells was seen. The theca externa of preantral immature follicles and hemorrhagic cysts showed strong expression of alpha-SMA. CONCLUSIONS: ERalphaKO mice show hemorrhagic polycystic ovaries and hyperplasia of the theca externa. This study demonstrates that the ERalpha is the functional key to the pathogenesis of PCOS.

Of berberine and puerarin on dexamethasone-induced insulin resistance in porcine ovarian thecal cells / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore the direct contribution of dexamethasone (Dex) for insulin resistance inducing in thecal cells and effects of berberine (Ber) and puerarin (Pue).</p><p><b>METHODS</b>Ovarian thecal cells from porcine follicles were isolated and cultured in vitro. Insulin resistance of thecal cells was induced by Dex treatment for 48 h. Then the glucose utilization ratio of thecal cells was detected. Meanwhile, the effects of Ber and Pue on insulin signal transmission and steroid hormones synthesis were determined by RT-PCR and Western blotting.</p><p><b>RESULTS</b>(1) After being treated by Dex for 48 h, the [3-3H] -glucose uptake in cells was lowered by about half, and the glucose content in supernate increased for about 1/3. (2) The RT-PCR and Western blotting showed that levels of insulin receptor substrate-1 (IRS-1), mRNA and protein expression of glucose transporter 4 (GLUT4) lowered, peroxisome proliferator-activated receptor -gamma(PPARgamma) and cytochrome P450 17 hydroxylase (CYP17) mRNA or protein expression increased in the model cells. However, the changes of above insulin signal molecules and CYP17 expression were inversed significantly after treated with Ber and Pue for 48 h. (3) As compared with the control, in the model cells, levels of testosterone (T, microg/mL) was higher (0.82 +/- 0.20 vs 0.38 +/- 0.01, P < 0.05), while after Ber and Pue treatment it was 0.44 +/- 0.24 and 0.45 +/- 0.21 respectively, all lower than that in the model cells (P < 0.05). No significant change of serum progesterone was found in all groups (P > 0.05).</p><p><b>CONCLUSIONS</b>After insulin resistance has been induced, the androgen synthetic capacity of thecal cells enhanced significantly. Ber and Pue could lower the degree of insulin resistance and the androgen synthesis in the model cells, displaying the favorable prospect of the two insulin sensitizing agents for the treatment of polycystic syndrome.</p>

The expression of p57(kip2) in normal ovaries and ovarian tumors / 대한산부인과학회지

OBJECTIVES: The purpose of this study is to investigate the expression of CDK (Cyclin dependent kinase) inhibitor, p57(kip2) in human ovarian corpus luteum, benign and malignant ovarian tumors. METHODS: 46 women undergoing laparoscopic surgery or laparotomy for ovarian tumors were enrolled. Total 46 formalin-fixed, paraffin-embedded sections of corpus luteum, benign and malignant ovarian tumors were stained by immunohistochemistry for expression of p57(kip2). RESULTS: p57(kip2) was stained in theca cell of growing follicle but not induced in human corpus luteum. There was the expression of p57(kip2) in mature teratoma, immature teratoma and endometrioma but not in epithelial ovarian tumors. CONCLUSIONS: These results showed that p57(kip2) expression may be not important in luteinization of the ovary and seemed not to play a role in development of epithelial ovarian tumors. However, it may involve pathogenesis of mature teratoma, immature teratoma and endometrioma.

Polycystic Ovary Syndrome and Insulin Resistance / 대한산부인과학회지

Polycysytic Ovary Syndrome (PCOS) is a common endocrine disorder characterized by chronic anovulation and hyperandrogenism. The etiology of PCOS is complex and incompletely understood. Accumulating data conclude that hyperinsulinemia and hyperandrogenemia may cause hormonal abnormalities that lead to disturbance of ovarian function. Although insulin resistance is not a part of the diagnostic criteria for PCOS, its importance in its pathogenesis can not be ignored. Excess insulin is capable of stimulating steroidogenesis and therefore excessive androgen production occurs from the theca cell system. Recently, the effects of insulin sensitizer in PCOS patients are being reported and they include the improvement of menstrual pattern, improvement in hyperandrogenism, increased response in ovulation induction and prevention of cardiovascular diseases. Understanding the relation of PCOS and insulin resistance will offer an improvement in treatment of PCOS in the future.

Characterization of Genes Related to the Cell Size Growth and CCN Family According to the Early Folliculogenesis in the Mouse / 대한불임학회지

OBJECTIVES: Previously, we sought to compile a list of genes expressed during early folliculogenesis by using cDNA microarray to investigate follicular gene expression and changes during primordialprimary follicle transition and development of secondary follicles (Yoon et al., 2005). Among those genes, a group of genes related to the cell size growth was characterized during the ovarian development in the present study. METHODS: We determined ovarian expression pattern of six genes related to the cell size growth (cyr61, emp1, fhl1, socs2, wig1 and wisp1) and extended into CCN family (connective tissue growth factor/cysteine-rich 61/nephroblastoma-overexpressed), ctgf, nov, wisp2, wisp3, including cyr61 and wisp1 genes. Expression of mRNA and protein according to the ovarian developmental stage was evaluated by in situ hybridization, and/or semiquantitative reverse transcriptase polymerase chain reaction (RT-PCR), and immunohistochemistry, respectively. RESULTS: Among 6 genes related to the cell size growth, cyr61 and wisp1 mRNA was detected only in oocytes in the postnatal day5 mouse ovaries. cyr61 mRNA expression was limited to the nucleolus of oocytes, while wisp1 was expressed in the cytoplasm and nucleolus of oocytes, except nucleus. cyr61 mRNA expression, however, was found in granulosa cells from secondary follicles. The rest 4 genes in the cell size growth group were detected in oocytes, granulosa and theca cells. Cyr61 and Wisp1 proteins were expressed in the oocyte cytoplasm from primordial follicle stage. Especially, Cyr61 protein was detected in pre-granulosa cells, Wisp1 protein was not. By using RT-PCR, we evaluated and decided that Cyr61 protein is produced by their own mRNA in pre-granulosa cells that was not detected by in situ hybridization. cyr61 and wisp1 genes are happen to be the CCN family members. The other members of CCN family were also studied, but their expression was detected in oocytes, granulose and theca cells. CONCLUSIONS: We firstly characterized the ovarian expression of genes related to the cell size growth and CCN family according to the early folliculogenesis. Cyr61 protein expression in the pre-granulosa cells is profound in meaning. Further functional analysis for cyr61 in early folliculogenesis is under investigation.

Distribution and Role of Ovarian Follicle Macrophage in Rat Ovarian Follicular Atresia / 대한산부인과학회지

OBJECTIVE: Ovarian follicular atresia is initiated from ovarian granulosa cell apoptosis and macrophages exert their effects directly and/or indirectly on follicular atresia by phagocytosis of apoptotic bodies and secretion of various cytokines. In spite of the abundant data on ovarian macrophages, the presence of these cells within the follicles (i.e., among granulosa cells) remains controversial and the elimination methods of apoptotic bodies of atretic follicles, and the time and methods of penetration of macrophages into the follicles are not known completely. The aim of the present study is to demonstrate the presence of macrophage within the ovary as related to follicular atresia and the process of elimination of apoptotic granulosa cells by light and electron microscopy. METHODS: Using rat ovaries, immunohistochemical studies with rat macrophage monoclonal antibody ED1 for macrophages, and light and transmission electron microscopic observations were performed. RESULTS: In the rat, follicular atresia was initiated by the granulosa cell apoptosis which occured randomly within the all granulosa layers. Macrophages were observed within normal follicles, in antrum, granulosa and theca cell layers of atretic follicels, in interstium and in corpus luteum. Ultrastructurally, apoptotic granulosa cells showed characteristics, pyknotic nucleus and apoptotic body formation. Apoptotic bodies were eliminated by intact neighboring granulosa cells and macrophages. Intact granulosa cells ingested apoptotic bodies transiently, soon after they fell into the apoptosis. Finally, apoptotic bodies and degenerating oocytes were phagocytosed by macrophages. Macrophages entered the ovarian follicle at the time of initiation of granulosa cell apoptosis, and migrated with the progression of apoptosis. By elimination of theca cells, macrophages contributed the completion of follicular atresia. CONCLUSION: This study demonstrates both intact neighboring granulosa cells and macrophages in the elimination of apoptotic bodies in atretic follicles of the rat ovary. Macrophages are present within normal follicles, in atretic follicles such as antrum, granulosa and theca cell layers and in corpus luteum but are in different appearances according to their location in ovary. A number of macrophages appearing in atretic follicles and in corpora lutea suggest a role for macrophages in follicular atresia and corpus luteum differentiation. The function of macrophage according to their location in follicular development should be further studied.