Evaluation of fertility and embryo implantation in rats after the oral administration of Salvia officinalis (sage) extract / Evaluación de la fertilidad y la implantación de embriones en ratas después de la administración oral de extracto de Salvia officinalis (salvia)

SUMMARY: In Saudi Arabia, it is widely believed that women with reproductive problems can use the extract of the sage plant as a tea drink. This study was conducted to investigate the effects of this herb on the fertility of female rats and embryo implantation. Forty-eight Wistar virgin female rats were divided into four groups at random, with 12 rats in each group. The control group received distilled water orally. The three treatment groups received different concentrations of sage extract: 15, 60, or 100 mg/kg for 14 days before mating, then mated with a male and sacrificed on the 7th day of gestation, the uterine horns removed, and photographed. The total body weight of mothers, weight of uteri and ovaries and number of fetuses were determined. Ovarian and uteri tissues were cut into 5 µ sections and stained with hematoxylin and eosin. Serum FSH, LH were determined by the ELISA method. The present study showed that low dose of sage (15 mg/kg) have no effects on serum concentration levels of FSH and LH hormones, also has no effect on the number of growing follicles. The present study showed a significant differences (P≤0.05) in body weight, ovary and uterus weight in the groups treated with high doses of Salvia officinalis as compared to control group. Also a significant differences (P≤0.05) found in FSH, LH hormones. Histological study showed overall histomorphological structural configurations including growing and matured graafian follicular countable changes, besides a number of corpora lutea and regressed follicles in the treated groups with high doses of Salvia officinalis as compared to control group. The researchers concluded that the extract of the sage plant with high doses can stimulate the growth graafian follicles and improve fertility in female rats.

RESUMEN: En Arabia Saudita, se cree ampliamente que las mujeres con problemas reproductivos pueden usar el extracto de la planta de salvia como bebida de té. Este estudio se realizó para investigar los efectos de esta hierba sobre la fertilidad de las ratas hembra y la implantación del embrión. Se dividieron cuarenta y ocho ratas hembra vírgenes Wistar en cuatro grupos al azar, con 12 ratas en cada grupo. El grupo control recibió agua destilada por vía oral. Los tres grupos de tratamiento recibieron diferentes concentraciones de extracto de salvia: 15, 60 o 100 mg/kg durante 14 días antes del apareamiento, luego se aparearon con un macho y se sacrificaron el día 7 de gestación, se extrajeron los cuernos uterinos y se fotografiaron. Se determinó el peso corporal total de las madres, el peso del útero y los ovarios y el número de fetos. Los tejidos ováricos y uterinos se cortaron en secciones de 5 µ y se tiñeron con hematoxilina y eosina. FSH sérica, LH se determinaron por el método ELISA. El presente estudio mostró que dosis bajas de salvia (15 mg/kg) no tienen efectos sobre los niveles de concentración sérica de las hormonas FSH y LH, tampoco tienen efecto sobre el número de folículos en crecimiento. El presente estudio mostró diferencias significativas (P≤0,05) en el peso corporal, peso de ovario y útero en los grupos tratados con altas dosis de Salvia officinalis en comparación con el grupo control. También se encontraron diferencias significativas (P≤0,05) en las hormonas FSH, LH. El estudio histológico mostró configuraciones estructurales histomorfológicas generales que incluyen cambios contables en los folículos maduros (de Graaf) y en crecimiento, además de una cantidad de cuerpos lúteos y folículos en regresión en los grupos tratados con altas dosis de Salvia officinalis en comparación con el grupo de control. Los investigadores concluyeron que el extracto de la planta de salvia en altas dosis puede estimular el crecimiento de los folículos maduros y mejorar la fertilidad en ratas hembra.

Insuficiencia ovárica primaria en adolescentes: revisión de la literatura a propósito de un caso clínico / Primary ovarian insufficiency in adolescents: review of the literature regarding a clinical case

El objetivo de este manuscrito es realizar una revisión y actualización de la literatura de la insuficiencia ovárica primaria (IOP) en población adolescente, a partir del diagnóstico, manejo y seguimiento de un caso clínico. La insuficiencia ovárica primaria se define como la menopausia en una mujer antes de los 40 años, acompañada de amenorrea, hipogonadismo hipergonadotrópico e infertilidad. Su prevalencia varía entre 1 a 2%, y en mujeres menores de 20 años su prevalencia es un caso de cada 10,000. Aunque se sabe que muchas afecciones pueden llevar a una IOP, la más común es la causa idiopática. La presentación clínica es diversa, y varios trastornos diferentes pueden también, llevar a esta condición. CASO CLÍNICO: Se presenta el caso de una adolescente de 17 años, previamente sana, con historia de amenorrea secundaria, no embarazada, con examen físico general y ginecológico normal. Se solicita estudio analítico complementario resultando con niveles de hormona folículo estimulante (FHS), estradiol (E2) y hormona antimülleriana (AMH) compatibles con una insuficiencia ovárica como la observada en la posmenopausia. Se inicia terapia hormonal (TH) clásica con estradiol y progesterona, siendo posteriormente reemplazada por anticoncepción hormonal combinada (AHC) oral, coincidente con el inicio de vida sexual, con respuesta favorable y sangrados regulares. La IOP tiene graves consecuencias para la salud incluyendo trastornos psicológicos como angustia, síntomas depresivos o depresión, infertilidad, osteoporosis, trastornos autoinmunes, cardiopatía isquémica, y un mayor riesgo de mortalidad. La enfermedad de Hashimoto es el trastorno autoinmune más frecuente asociado a la IOP. Su tratamiento y diagnóstico deben establecerse de forma precoz para evitar consecuencias a largo plazo. La terapia con estrógenos es la base del tratamiento para eliminar los síntomas de la deficiencia de estrógenos, además de evitar las consecuencias futuras del hipogonadismo no tratado. También el manejo debe incluir los siguientes dominios: fertilidad y anticoncepción, salud ósea, problemas cardiovasculares, función psicosexual, psicológica y neurológica, informando a los familiares y a la paciente sobre la dimensión real de la IOP y la necesidad de tratamiento multidisciplinario en muchos casos. CONCLUSIÓN: El caso presentado, pese a ser infrecuente, permite abordar de manera sistematizada el diagnostico de IOP y evaluar alternativas de manejo plausibles para evitar graves consecuencias en la salud, así como conocer respuesta clínica y de satisfacción de la adolescente.

The objective of this manuscript is to review and update the literature on primary ovarian insufficiency (POI) in an adolescent population, based on the diagnosis, management and follow-up of a clinical case. Primary ovarian insufficiency is defined as menopause in a woman before the age of 40, accompanied by amenorrhea, hypergonadotropic hypogonadism, and infertility. Its prevalence varies between 1 to 2%, and in women under 20 years of age its prevalence is one case in every 10,000. Although it is known that many conditions can lead to POI, the most common is the idiopathic cause. The clinical presentation is diverse, and several different disorders can also lead to this condition. CLINICAL CASE: The case of a 17-year-old adolescent, previously healthy, with a history of secondary amenorrhea, not pregnant, with a normal general physical and gynecological examination is presented. A complementary analytical study is requested, resulting in levels of follicle stimulating hormone (FHS), estradiol (E2) and anti-müllerian hormone (AMH) compatible with ovarian insufficiency such as that observed in postmenopause. Classic hormonal therapy (HT) with estradiol and progesterone was started, later being replaced by combined hormonal contraception (CHC), coinciding with the beginning of sexual life, with a favorable response and regular bleeding. POI has serious health consequences including psychological disorders such as distress, depressive symptoms or depression, infertility, osteoporosis, autoimmune disorders, ischemic heart disease, and an increased risk of mortality. Hashimoto's disease is the most common autoimmune disorder associated with POI. Its treatment and diagnosis must be established early to avoid long-term consequences. Estrogen therapy is the mainstay of treatment to eliminate the symptoms of estrogen deficiency, in addition to avoiding the future consequences of untreated hypogonadism. Management should also include the following domains: fertility and contraception, bone health, cardiovascular problems, psychosexual, psychological and neurological function, informing family members and the patient about the real dimension of POI and the need for multidisciplinary treatment in many cases. CONCLUSION: The case, although infrequent, allows a systematic approach to the diagnosis of POI and evaluate plausible management alternatives to avoid serious health consequences, as well as to know the clinical response and satisfaction of the adolescent.

Buchholzia coriacea (wonderful kola) seeds induce male reproductive toxicity by suppressing the pituitary-gonadal axis in Wistar rats

The methanolic extract of Buchholzia coriacea seeds (MEBC) has been reported to induce male reproductive toxicity by decreasing sperm parameters and fertility index. To elucidate the possible mechanism(s), the effects of graded doses of MEBC on sex hormones and sperm profile were investigated in this study. The MEBC (e.g., 50, 200, 400, and 600 mg/kg) was administered daily (p.o.) to male Wistar rats for 6 weeks, while a concurrent control group received distilled water (vehicle). Then, the animals were sacrificed under sodium pentobarbital anaesthesia. Weights of organs were recorded, and the sperm profile was determined microscopically. Testosterone, luteinizing hormone (LH), and follicle stimulating hormone (FSH) were assayed from the obtained serum using the ELISA technique. Sperm motility was significantly reduced by MEBC (i.e., 50 and 200 mg/kg), and sperm count reduced in all treated groups in a dose-dependent manner compared with that of the control. Serum testosterone, LH, and FSH decreased in treated rats. A histopathological examination of testes showed a considerable depletion and necrosis of the epithelium of seminiferous tubules. The result suggests that Buchholzia coriacea seeds induce male reproductive toxicity by suppressing the pituitary-gonadal axis.

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)

Produção de embriões em ovinos Morada Nova e Somalis Brasileira / Embryo production in Morada Nova and Somalis Brasileira ewes

Superovulatory response and embryo yield in 19 Morada Nova and 20 Somalis Brasileira ewes was analyzed. All animals were synchronized with the insertion of an intravaginal device (CIDR®) on Day 0, replaced by a new device on Day 7, which remained in place until Day 14 and superovulated with 133mg of porcine FSH (pFSH) in decreasing doses at 12h intervals from Day 12 until Day 15 of the treatment, and a single dose of equine chorionic gonadotropin (eCG, 200UI) on Day 14 (i.e., administered in CIDR removal). Fifty hours after CIDR® removal, females were inseminated by laparoscopy. All embryos were recovered by laparotomy 5 days after insemination. Sheep which responded to the superovulation protocol (P>0.05) included 74% of the Morada Nova ewes and 50% of the Somalis Brasileira ewes. Morada Nova showed better results (P<0.05) than Somalis Brasileira in number of ovulations (15.38 ± 5.24 vs. 10.56 ± 2.83), total structures (11.00 ± 7.55 vs. 3.33 ± 1.94) and embryo yields (6.79 ± 5.35 vs. 2.90 ± 2.18). Despite the high fertilization rate, degenerate embryo rate was high for both breeds, with an overall rate of 39% (57/145). In conclusion, superovulatory response and embryo yields in Morada Nova ewes were considered sufficient to justify the use of this procedure in genetic resources conservation programs. However, improvements to embryo quality and control of precocious regression of corpus luteum are necessary to produce better results in the MOET program, with minimal variations and maximum embryo yield in Morada Nova and Somalis Brasileira ewes.(AU)

Redução da resposta superovulatória de fêmeas bovinas superestimuladas com FSH em doses split / Reduced superovulatory response of bovine females overstimulated with FSH in split doses

Objetivou-se avaliar a superovulação (SOV) de vacas zebuínas, utilizando protocolo convencional ou protocolo com número menor de aplicações e similar dosagem (dose split). Utilizaram-se 16 fêmeas (total 32 SOV), com idade entre 17-42 meses e escore de condição corporal 2,5-4 (escala de 1-5), em delineamento tipo cross-over. No início do tratamento (D0), os animais receberam um dispositivo de progesterona e 2mg de benzoato de estradiol. As fêmeas do grupo convencional receberam 250UI de FSH/LH divididas em oito doses decrescentes administradas em intervalos de 12h (FSH/LH no D4, D5, D6 e D7 no período da manhã e tarde, nas respectivas dosagens: 50,0 UI; 37,5 UI; 25,0 UI; 12,5 UI). No D7 pela manhã, as fêmeas foram tratadas com 150μg de D+cloprostenol, e a remoção da progesterona foi realizada no D7 à tarde. As fêmeas do grupo split também receberam 250 UI de FSH/LH. No D4 de manhã, administraram-se 62,5 UI de FSH/LH via IM e 125 UI por via SC. Quarenta e oito horas após (D6) administraram-se 62,5 UI via SC e na manhã do D7 foi removida a progesterona e aplicaram-se 150μg de D+cloprostenol. As fêmeas de ambos os grupos receberam 50μg de análogo de GnRH no D8 pela manhã e foram inseminadas 12 e 24 horas após. No D15 realizou-se a colheita dos embriões em ambos os tratamentos. Avaliou-se a resposta superovulatória pela contagem do número de folículos e CLs de cada ovário, com auxílio de ultrassom. Todas as variáveis foram submetidas ao teste T de Student para amostras pareadas. Houve diferença (P<0,05) na quantidade de folículos acima de 8mm no D8 (9,06±4,54 e 5,50±4,59); número de CLs no dia da colheita (8,12± 3,26 e 4,69±3,46), número de embriões totais (6,69±3,05 e 3,37±2,50) e de embriões viáveis (5,25±2,29 e 2,37±1,78) nas vacas do grupo convencional em relação às do split, respectivamente. Conclui-se que o protocolo split tem pior resposta superovulatória e de produção in vivo de embriões, em vacas zebuínas, quando comparado ao protocolo convencional...

The aim of this study was to evaluate the superovulation (SOV) response of zebu cows, using conventional protocol or other protocol with similar dose but smaller number of applications (split dose). 16 females (32 SOV), aged 17-42 months, and body condition score of 2.5-4 (1-5 scale) were used in randomized cross-over. At the start of treatment (D0), the animals received progesterone device and 2mg of estradiol benzoate. The females from the conventional group received 250 IU of FSH/LH divided into eight decreasing doses administered at intervals of 12 hours (FSH/LH in D4, D5, D6 and D7, with their respective strengths: 50.0 IU, 37.5 IU, 25.0 IU, 12.5 IU). In D7, the females were treated with 150μg of D+cloprostenol, and the removal of progesterone device was held in the afternoon. The females from the split group also received 250 IU of FSH/LH. In the morning D4 was administered in 62.5 IU FSH/LH IM and 125 IU subcutaneously. Forty eight hours later (D6) 62.5 IU was administered subcutaneously in the morning and on D7, the progesterone device was removed and 150 μg of D+cloprostenol was applied. The females in both groups were given 50 μg of GnRH in the morning and on D8 were inseminated after 12 and 24 hours. On D15 the embryo collection was performed in both treatments. The evaluation of superovulatory response was done by counting the number of follicles and corpus luteum (CL) in each ovary, with the aid of ultrasound. The variables were assessed by Student's t test for paired samples...

Toxic effects of lead exposure during gestation and lactation on testicular development in albino rat offspring: a hormonal and histopathological study

This study aims at exploring the effect of lead exposure during pregnancy and lactation on the rat testis, which was evaluated via light and electron microscopic examination of testicular tissue, besides estimation of serum follicle stimulating hormone [FSH], luteinizing hormone [LH] and testosterone [TT] levels. Dams were intoxicated with 10 mg/kg body weight [BW]/day lead acetate throughout the gestational period and lactation. Pups were sacrificed on postnatal [PN] days 1 and 21. The weights of the body and testes, as well as serum gonadotropins and TT concentrations were significantly reduced in pups from lead-intoxicated mothers compared with their controls. Light microscopic examination of testicular samples from lead-exposed pups revealed thin-walled seminiferous tubules with significant reduction in their size. In addition, multiple vacuoles were seen within the tubules and in the interstitium, and the interstitial spaces appeared enlarged with reduced number of Leydig cells. Apoptotic bodies were found among the basal parts of the spermatogenic epithelium. Examination of testicular tissue from the same group by electron microscopy revealed more cellular details confirming the toxic effects of lead. The altered seminiferous tubules showed multiple cytoplasmic vacuoles and precipitates along the nuclear membranes in Sertoli cells. Several seminiferous tubules showed apoptotic cells with heterochromatic nuclei and dense cytoplasm, whereas other tubules appeared ensheathed by single layers of myoid cells. Leydig cells revealed cytoplasmic vacuolations and irregular nuclei with chromatin masses. The results of this study indicate that lead intoxication affected the normal development of germinal cells and disrupted the testicular structure and endocrinal functions

Protocolo largo con análogos de GnRH versus protocolo corto con antagonistas: ¿existen diferencias en cuanto a los resultados de los ciclos de FIV-ICSI? / Protocol with GnRH analogues vs antagonists of GnRH: exist differences in the results of the IVF-ICSI cycles?

Objetivo: Valorar si existen diferencias en los resultados de los ciclos de FIV-ICSI en función del protocolo de estimulación empleado. Método: Estudio retrospectivo descriptivo de pacientes infértiles que fueron sometidas a ciclos de FIV-ICSI en el Hospital Universitario La Paz, entre los meses de enero y septiembre de 2010, comparando un protocolo largo de estimulación con análogos de GnRH vs un protocolo corto con antagonistas de GnRH. Las variables analizadas fueron: tasa de gestación, necesidad de cancelación del ciclo, dosis total de gonadotropinas requerida durante la estimulación, niveles de estradiol sérico el día de la administración de la hCG, número de folículos puncionados, complejos obtenidos, número de ovocitos maduros y de embriones conseguidos. Resultados: No hubo diferencias estadísticamente significativas en los resultados de los ciclos en función del protocolo de estimulación empleado, en ninguna de las variables analizadas. Conclusiones: Este estudio no encontró diferencias en los resultados de los ciclos de FIV-ICSI con relación al uso de análogos o antagonistas de GnRH. Es necesarios más estudios con mayores tamaños muestrales para definir qué tipo de pacientes serían subsidiarias de recibir cada tratamiento para conseguir resultados óptimos.

Aims: To assess if there exist any differences in the results of the IVF-ICSI cycles depending on the stimulation protocol employed. Methods: Retrospective descriptive study of infertile patients who underwent IVF-ICSI cycles at La Paz University Hospital, between January and September 2010, comparing sitmulation protocol with GnRH agonists vs antagonists of GnRH. The variables analyzed were pregnancy rate, cancellation rate, total dose of gonadotropin required for stimulation, serum estradiol levels on the day of hCG administration, number of follicles punctured, complexes obtained, number of mature oocytes and of embryos obtained. Results: No statistically significant differences where found in the results of cycles depending on the protocol of stimulation used in any of the variables analyzed. Conclusions: This study didn't find any difference in the outcome of IVF-ICSI cycles in relation to the use of GnRH agonists or antagonists. We need more studies with larger sample sizes to determine which is the best treatment to each patient in order to achieve optimal results.

[Assessment of serum LH, FSH /tumor fluid FSH, LH ratio in ovarian tumors]

As ovarian cancers are prevalent and have a high mortality rate, early diagnosis is important. Markers that detect ovarian malignancies earlier, include the levels of serum and ovarian cyst fluid gonadotropins. This study was performed in Imam Hossein hospital between 2008 and 2010 to determine the diagnostic value of the ratio of serum to ovarian cyst fluid gonadotropins in defining the type of ovarian tumors. This cross-sectional study was performed on patients operated because of ovarian mass. Their serum samples and cyst fluid were collected at the time of laparatomy and the histopathologic diagnosis was established after removal of the tumor. FSH and LH concentration were measured in serum and tumor fluid by Microparticle Enzyme Immunoassay technology in Imam Hossein laboratory and the ratios of serum FSH and LH to the cyst fluid gonadotropins were determined in four pathologic groups [simple cyst, benign, borderline and malignant]; statistical comparison was done with the Kruskal-Wallis and Mann-Whitney U-test. We studied 100 patients with ovarian masses; in 79 patients pathology was reported as epithelial tumors and simple cysts. The most common mass were simple cysts [48.1%], followed by benign tumors [27.8%], malignant tumors [16.5%] and borderline tumors [7.6%]. Serum FSH/ tumor fluid ratios were, 21.1 +/- 53.8 in simple cysts, 50.7 +/- 174.9 in benign tumors, 1.5 +/- 0.7 in borderline tumors and 1.9 +/- 1.9 in malignant tumors [P<0.001]. Serum LH/tumor fluid ratios were 12.1 +/- 26.5 in simple cysts, 20.0 +/- 29.1 in benign tumors, 2.6 +/- 2.3 in borderline tumors and 2.8 +/- 3.7 in malignant tumors [P<0.002]. Our findings suggest that serum/tumor fluid gonadotropins ratio can be used as a tumor marker for identification of ovarian tumors

GnRH Agonist Therapy to Protect Ovarian Function in Young Korean Breast Cancer Patients

The increased survival of patients with breast cancer has given rise to other problems associated with the complications of chemotherapy. One major complication is premature ovarian failure, an especially harmful outcome for women of reproductive age. This study was performed to evaluate the efficacy of GnRH agonist (GnRHa) treatment on protecting ovarian function in young breast cancer patients (30.59+/-5.1 yr) receiving chemotherapy after surgery. Twenty-two women were enrolled and given subcutaneous injections of leuprolide acetate (3.75 mg) every 4 weeks during chemotherapy. Follow-up laboratory tests (luteinizing hormone [LH], follicle stimulating hormone [FSH], and estradiol) were performed 1, 3, and 6 months after chemotherapy. Menstruation patterns and clinical symptoms were followed up for a mean duration of 35.6+/-1.7 months. FSH and LH levels were normal in all patients 6 months after completing chemotherapy (8.0+/-5.3, 4.4+/-2.7 mIU/mL, respectively). During follow-up, none of the patients complained of menopausal symptoms and 81.8% experienced recovery of menstruation. This report is the first trial of GnRHa as a treatment modality to protect ovarian function during adjuvant chemotherapy in young Korean breast cancer patients.

Immunodetection of Luteinizing Hormone (LH), Follicle-Stimulating Hormone (FSH), Thyroid Stimulating Hormone (TSH) and Prolactin (PRL) in Brachionus calyciflorus (Rotifera: Monogononta)

The endocrine system controls and coordinates behavioral, biochemical, and physiological processes through signal mechanisms using neuropeptides or products of neurosecretory cells. Among invertebrates, this system is poorly studied in rotifers, in which estrogens and androgens significantly affect sexual reproduction. This is the first report of the presence of the Luteinizing Hormone (LH), Follicle-Stimulating Hormone (FSH), Thyroid Stimulating Hormone (TSH) and Prolactin (PRL) in rotifers. Analyses included the avidin-biotin-peroxidase complex method with primary antibodies LH (Anti-Rat LH serum for RIA), PRL (Anti-Rat PRL serum for RIA), FSH (Anti-Rat FSH serum for RIA) and TSH (Anti-Rat TSH serum for RIA). These hormones were found in females, males and parthenogenetic and sexual eggs of the freshwater Brachionus calyciflorus. The immunoreactivity of FSH, LH, TSH and PRL in females was observed in: ovaries, cerebrum, mastax, stomach, lorica, and the stomach gland. However, in males LH was observed only at the trochal disk and cerebrum. The hormones FSH, TSH and PRL, were observed in testicles, contractil vesicles, and cementary gland of males. Regarding amictic or parthenogenetic eggs, the hormones LH, FSH, TSH, and PRL were located mainly in the micromeres, and the staining in the macromeres was weak. On the other hand, in the mictic or sexual eggs the inner shell is stained for the hormones PRL and LH, opposite to the staining of FSH and TSH, located mainly in the embryo. In general, immuno-reactivity was observed in areas important for the reproductive, excretory, digestive and developmental processes. Rev. Biol. Trop. 57 (4): 1049-1058. Epub 2009 December 01.

Se logró detectar la presencia de las hormonas: Hormona Luteinizante (LH), Hormona Folículo Estimulante (FSH), Hormona Estimulante de la Tiroides (TSH) y Prolactina (PRL) en Brachionus calyciflorus siendo el primer reporte de la presencia de dichas hormonas en rotíferos. Estas hormonas fueron identificadas por un método inmunológico-histológico-químico usando el complejo avidina-biotina- peroxidasa con los siguientes anticuerpos primarios: LH (Anti-Rata LH suero para RIA), PRL (Anti-Rata PRL suero para RIA), FSH (Anti-Rata FSH suero para RIA) y TSH (Anti-Rata TSH en suero para RIA). Estas hormonas se encontraron en las hembras, machos, huevos partenogenéticos y huevos sexuales del rotífero dulceacuícola B. calyciflorus. La reactividad inmunológica de FSH, LH, PRL y TSH en las hembras se observó en ovarios, cerebro, mástax, estómago, lorica, y la glándula del estómago. Sin embargo, en machos, la LH se observó sólo en el disco trocal y cerebro mientras que las hormonas FSH, PRL y TSH, se observaron en testículos, vesícula contráctil, y la glándula cementaria. En cuanto a los huevos partenogenéticos o amícticos, las hormonas LH, FSH, TSH, y PRL, se encontraron principalmente en los micrómeros, y en los macrómeros la tinción es débil. Por otra parte, el huevo sexual o míctico muestra reactividad inmunológica en la cubierta interior del huevo para las hormonas LH y PRL, lo contrario para FSH y TSH, las cuales se observaron principalmente en el embrión. La reactividad inmunológica fue observada, en general, en áreas importantes para los procesos reproductivos, excretorios, digestivos y del desarrollo.

Evaluación por inmunohistoquímica de la expresión de hormonas hipofisiarias y del marcador de proliferación celular Ki-67 en tejido de adenomas causantes de acromegalia / Immunohistochemistry for pituitary hormones and Ki-67 in growth hormone producing pituitary adenomas

Background: Growth hormone (GH) producing adenomas, frequently express several hormones. This condition could confer them a higher proliferative capacity. Ki-67 is a nuclear protein antigen that is a marker for proliferative activity. Aim: To measure the immunohistochemical hormone expression in pituitary adenomas, excised from patients with acromegaly. To determine if the pluríhormonal condition of these adenomas is associated with a higher proliferative capacity, assessed through the expression of Ki-67. Material and methods: Forty one paraffin embedded surgical samples of pituitary adenomas from patients with acromegalia were studied. Immunohistochemistry for GH, prolactin (PRL), follicle stimulating hormone (FSH), luteinizing hormone (LH), thyroid stimulating hormone (TSH), adrenocorticotropin (ACTH) and for the expression of Ki-67 was carried out. Results: All samples were positive for GH. Twenty seven had positive staining for PRL, 12 had positive staining for glycoproteic hormones and 11 for PRL and one or more glycoproteic hormones. Mean staining for Ki-67 was Z.6±3.3 percent. There were no differences in the expression of this marker between mono or pluríhormonal tumors. The expression was neither associated with extrasellar extensión. Conclusions: Half of GH producing pituitary adenomas are pluríhormonal. There are no differences in the expression of Ki-67 between mono and plurihormonal adenomas.

LH/FSH ratio: a better marker of secondary amenorrhea in patients from eastern Nepal.

FSH, LH and prolactin (PRL) levels were assessed by ELISA in 50 cases with secondary amenorrhea and 52 age and sex-matched healthy controls from eastern Nepal. Cases were diagnosed by differential diagnosis, and data were analyzed using standard statistical tools. Early stage (3-6 months) and long standing (> 6 months) secondary amenorrhea had no effect (p > 0.05) in hormonal parameters studied. Pulse, SBP, DBP, weight, height, age of menarche, cycle interval and duration of flows were homogenous (p > 0.05) in patients and controls. Median age of menarche, median cycle interval and median duration of flows in healthy subjects were 14 years, 30 and 4 days respectively. FSH in cases (15.38 +/- 7.24 mU/ml) was significantly elevated (p < 0.01) as compared to controls (9.38 +/- 6.34 mU/ml). LH in cases (35.44 +/- 24.35 mU/ml, median 36.5 mU/ml) was significantly (p < 0.01) elevated by almost 5 times of its mean value and 9 times of its median value as compared to that of controls (7.58 +/- 6.604 mU/ml, median 4.2 mU/ml). LH/ FSH ratio in cases (2.44 +/- 1.73, median 2.00) was significantly higher (p < 0.01) as compared to controls (0.82 +/- 0.42, median 0.76). FSH e"12 mU/ml, LH e"10 mU/ml and LH/FSH ratio e"1 cut offs were significantly associated (p = 0.000 in each) with the cases as revealed by chi-square analysis, and LH/FSH ratio e"1 (Sensitivity = 84.0%, specificity = 77.0%) was found to be a stronger marker of secondary amenorrhea. As the elevation of LH was more pronounced than that of FSH, this study hints towards possible LH receptor mutation, which is generally found in premature ovarian failure (POF). Diagnosis of cases in this region may need a new cut off level for POF, as the elevation of FSH itself was not as pronounced as reported by other workers.

Serum estradiol and follicle-stimulating hormone levels in Thai women post total abdominal hysterectomy and bilateral oophorectomy using oral 17 beta-estradiol.

OBJECTIVE: To assess the difference of serum estradiol (E2) and follicle stimulating hormone (FSH) levels in Thai women post total abdominal hysterectomy and bilateral oophorectomy, before and after using a daily dose of 2 mg oral 17 beta-estradiol. STUDY DESIGN: Experimental study (before and after treatment). SETTING: Menopause Clinic, Department of Obstetrics and Gynecology, Faculty of Medicine, Chulalongkorn University. SUBJECTS: Thirty-five women who had undergone total abdominal hysterectomy and bilateral oophorectomy at Chulalongkorn Hospital 1 week previously due to benign gynecologic conditions were recruited in the study. Body mass index was 20-25 kg/m2. These women had no contraindication for using hormonal replacement therapy and no history of any hormonal intake in the past. INTERVENTION: All subjects were assigned to receive a daily dose of 2 mg oral 17 beta-estradiol applied at bedtime (8.00 p.m.). MAIN OUTCOME MEASURES: Serum E2 and FSH were measured before and after the study at weeks 4, 8 and 12, 12-14 hours after oral application. The hormonal measurement was performed using the time-resolved fluoroimmunoassay (FIA) method. RESULTS: Five cases were excluded, three cases due to poor compliance which was less than 85 per cent and two cases due to loss to follow-up. Of the remaining 30 cases, the mean age and body mass index were 43.03 +/- 4.58 years and 22.72 + 1.86 kg/m2, respectively. Serum E2 level significantly increased from baseline value at 4, 8 and 12 weeks (median of E2 value at 0, 4, 8 and 12 weeks: 20.00, 22.50, 324.65 and 355.35 pmol/L, p<0.001). On the other hand, there was no significant difference in the FSH serum level (median of FSH value at 0, 4, 8 and 12 weeks: 18.65, 18.40, 18.60 and 20.35 IU/L, p=0.517). CONCLUSION: A daily dose of 2 mg oral 17 beta-estradiol taken at bedtime (8.00 p.m.) for 12 weeks increased the serum E2 to the follicular phase level of the normal menstrual cycle. On the other hand, there was no significant difference in the FSH serum level.

Elevada incidencia de hiperestrogenemia y de dislipidemia en un grupo de hombres infértiles / Hyperestrogen and dyslipidemia high-incidence in a group of infertile men

Objetivo. Describir algunas alteraciones metabólicas y endocrinas en un grupo de hombres de parejas estériles. Métodos. Incluimos 106 hombres en forma consecutiva, con el propósito de analizar sus muestras séricas para la determinación por duplicado de hormona luteinizante (LH), hormona foliculoestimulante (FSH), estradiol (E2), testosterona libre (TL), 17alfa-hidroxiplrogesterona (17OHP), androstenediona (A), sulfato de dehidroepiandrosterona (S-DHEA), prolactina (PRL), insulina, glucosa, colesterol total y triglicéridos. Resultados. Se encontró una incidencia de dislipidemia de 65 por ciento (hipercolesterolemia aislada, hipertrigliceridemia aislada o ambas) en el grupo, donde 80 por ciento de los pacientes son menores de 40 años. No se encontró correlación con obesidad, sobrepeso o con alguna alteración endócrina o tipo de alteración espermática. En el grupo completo se encontró una correlación positiva entre E2 y FSH (r = 0.67, p < 0.0001) que continuó siendo significativamente en el subgrupo de hombres con hiperestrogenismo (n = 27, r = 0.68, p < 0.0001) pero no en el subgrupo de hombres con estradiol sérico normal (n = 79, r = 0.10, NS). Otras alteraciones: obesidad en 18 por ciento, sobrepeso 30.2 por ciento, diabetes mellitus 4.7 por ciento, intolerancia a la glucosa 15 por ciento, hipertensión arterial 26 por ciento (14/53), hipogonadismo hipergonadotrófico 3.7 por ciento. Conclusión. De los 106 pacientes consecutivos, sólo ocho (8.4 por ciento) no tuvieron alguna de las alteraciones endócrinas o metabólicas descritas. Estos datos son significativos ya que 83 por ciento de los pacientes son menores de 40 años. Es novedosa la correlación positiva encontrada entre el E2 y la FSH cuando los niveles séricos de estradiol exceden de 50 pg/mL.

Quality control in clinical pathology

The reliability and validity of any test related to clinical pathology is the key parameter for its integrity. This can be monitored by using control samples having known values of the particular analyte, and determining the precision in reproducibility of these samples. The quality control programme used in our laboratory is discussed, through which extra costs involved in purchasing control samples have been cut without compromising upon the integrity of the results

Esteróides anabolizantes e suas alteraçöes em análises clínicas / Anabolic steroids and its alterations in clinical chemistry

Os esteróides anabolizantes säo compostos sintéticos semelhantes à testosterona e apresentam efeitos anabolizantes e androgênicos. Os esteróides anabolizantes possuem alguns usos terapêuticos, mas apresentam muitos efeitos tóxicos: alguns reversíveis, como diminuiçäo na produçäo de espermatozóides e impotência; outros irreversíveis, como ginecomastia e calvície. Essas substâncias säo utilizadas por jovens e atletas, em dosagens muito elevadas, na busca de uma melhora no desempenho e aparência física. Laboratorialmente, é possível observar alteraçöes bioquímicas e hematológicas, entre elas diminuiçäo do HDL colesterol, aumento de enzimas hepáticas, glicose e hematócrito, causadas pelo uso destas substâncias. Por isso, é importante conhecer e identificar essas alteraçöes, para alertar médicos sobre sua possível causa, o que pode gerar mudanças no diagnóstico e tratamento de pacientes

Antiovarian antibody in premature ovarian failure.

Premature ovarian failure is a syndrome consisting of primary or secondary amenorrhoea, hypergonodotropiremia and hypoestrogenemia in women under the age of 40. An autoimmune mechanism was suggested as possible etiology when Vallolton and Forbes in 1966-67 found antibodies to the cytoplasm of rabbit ova in 29 of 232 tested sera. Immune mechanism in the pathogenesis of premature ovarian failure (POF) is suggested by association of autoimmune phenomenon with POF in some cases and demonstration of circulating antibodies to ovary in serum samples from women with POF. The incidence of presence of antiovarian antibody of POF patients has been reported earlier. Evidence of autoimmunity is present in 18-92% of patients with POF. In the present study we have studied 18 cases of POF without any overt manifestation of autoimmune disorder but the antiovarian antibody was detected, with the idea that this autoantibody might be the cause of ovarian dysfunction which is evident in POF. Presence of antiovarian antibody in 16.67% cases with POF in our study that ovarian antibodies may play a role in or reflect an autoimmune process responsible for the development of POF.

Interpreting immunoassays

Immunoassay are carried out to quantify body fluids, and form an integral part of the clinical diagnosis. However, normal ranges of laboratories differ and the same value may be normal for one laboratory and pathological for another. Thus it becomes extremely important to correlate the clinical picture with the chemical pathology profile

Hormonal control of spermatogenesis in adult albino rat; ultrastructural, endocrinological 1-fsh and testosterone

A quantitative and qualitative analysis of germ cell populations in normal, hypophysectomized [Hx] and Hx-hormone treated animals was undertaken during the periods of regression that were characterized as intermediate between short-term and long-term regression of the testis. Ten groups of adult rats were utilized and administered either follicle-stimulating hormone [FSH] or testosterone [T] in different combinations. Flutamide and casodex were used to compete with and rogen binding to receptors in Hx animals where it is known that small amounts of and rogen are secreted in spite of the lack of a pituitary gl and. FSH and T significantly increased the testis weight as compared with Hx controls, but FSH and T were the most effective either singly or in combination. A novel assay for the determination of the numbers of viable germ cells was used in these groups to determine the cellular sites of FSH and T action