Estudio prospectivo randomizado comparando la via de administración subcutáneas vs. la via intramuscular de depósito de acetato de leuprolide en técnicas de reproducción asistida / Randomized study comparing subcutaneous administration of vial vs. the intramuscular vial of acetate in techniques assited reproduction

Comparar los resultados en técnicas de reproducción asistida (ART) empleando dos vias diferentes de administración de análogos de GnRH (diario vs. depósito9

Estudio prospectivo randomizado comparando la via de administración subcutáneas vs. la via intramuscular de depósito de acetato de leuprolide en técnicas de reproducción asistida / Randomized study comparing subcutaneous administration of vial vs. the intramuscular vial of acetate in techniques assited reproduction

Comparar los resultados en técnicas de reproducción asistida (ART) empleando dos vias diferentes de administración de análogos de GnRH (diario vs. depósito9(AU)

Mass spectrometry, high performance liquid chromatography, and brain peptides.

This paper is a personal recollection of some of the events and research that surrounded the amino acid sequence determination of the hypothalamic releasing factor, TRF (now known as TRH), by mass spectrometry (MS), and the development of reverse phase high performance liquid chromatography (RP-HPLC) MS and tandem MS (MS/MS) methods for the qualitative and quantitative analysis of native opioid neuropeptides in human pituitary tissue extracts.

Manejo simplificado de un programa de fertilización in vitro / Simplify management of an in vitro fertilization programme

El desarrollo de la fertilización in vitro ha acelerado significativamente el conocimiento de la fisiología reproductiva humana. Los resultados obtenidos han demostrado ser consistentes y reproductibles en diferentes centros de medicina reproductiva en casi todo el mundo. Es sabido que la repetición de la técnica se traduce en tasas acumuladas de embarazo de hasta 60 por ciento Sin embargo, en casos de falla del método, la mayor limitante a los nuevos intentos es, por un lado su costo económico y por otro las opciones quirúrgicas de la técnica. En este estudio se presentan los resultados de un programa que ha intentado simplificar la técnica mediante: el empleo de transferencias no quirúrgicas; el monitoreo ecográfico de la inducción de ovulación y la disminución de la dosis de agonistas de la hormona liberadora de gonadotropinas para la supresión del eje hipotálamo hipófisis. Los resultados en 47 de estos ciclos no muestran diferencias significativas, respecto de la literatura, en lo referente al porcentaje de madurez ovocitaria, la tasa de fertilización y clivaje. La tasa de embarazo por aspiración es de 28,6 por ciento y de 32,3 por ciento por transferencia. Concluimos que el usoi del ultrasonido vaginal como único elemento de monitoreo de la inducción de ovulación, y el uso casi exclusivo de transferencias no quirúrgicas no parecen alterar los resultados

Agonistas de GnRH en ginecología II / Agonists of GnRH in gynecology II

La presente revisión tiene como objeto describir las características químicas y farmacológicas de los análogos de GnRH y las indicaciones actuales de su uso en ginecología. Se realiza una revisión crítica del uso de análogos de GnRH en: Anticoncepción Reproducción asistida, Miomatosis uterina, Endometriosis, Poliquistosis ovárica, Pubertad precoz, Síndrome de tensión premenstrual y cáncer de mama.

Endocrine function in multiple sclerosis.

In 31 patients with multiple sclerosis (MS) the endocrine functions of the hypothalamus, the pituitary and several peripheral endocrine glands were assessed with a combined pituitary test; 3/31 patients had an endocrine disease: one primary hypothyroidism, one primary amenorrhea and one primary male hypogonadism. We found no patient with endocrine disease of the hypothalamus, the pituitary or the adrenals. However, the poststimulatory secretion of cortisol, growth hormone or thyroid-stimulating hormone was impaired in 7/31 patients, suggesting a possible preclinical endocrine insufficiency in these patients.

Agonistas de GnRH en ginecología I / Agonist of GnRH in gynecology I

La presente revisión tiene como objeto describir las características químicas y farmacológicas de los análogos de GnRh y las indicaciones actuales de sus uso en ginecología. Se realiza una revisión crítica del uso de análogos de GnRH en: Anticoncepción, Reproducción Asistida, Miomatosis Uterina, Endometriosis, Poliquistosis ovárica, Pubertad Precoz, Síndrome de Tensión Premenstrual y Cáncer de mama.

Effect of dietary energy, body condition and calf removal on pituitary gonadotropins, gonadotropin-releasing hormone (GnRH) and hypothalamic opioids in beef cows.

Beef cows (n = 64) were slaughtered to evaluate effects of dietary energy and calf removal (CR) on hypothalamic and adenohypophysial endocrine characteristics. From d 190 of gestation until parturition, cows received maintenance (ME; n = 32) or low (LE; n = 32) energy diets (ME = 100%, LE = 70% NRC recommendations). After parturition, half (n = 16) of each prepartum diet group received low (LE; n = 32) or high (HE = 130% NRC; n = 32) energy diets. At 30 d postpartum, cows were slaughtered 0 or 48 hr after CR. Hypothalami [preoptic area (POA), hypothalamus (HYP), stalk-median eminence (SME)] and pituitaries were collected. Basal and K(+)-induced release of GnRH from SME, and pituitary luteinizing hormone (LH) and follicle stimulating hormone (FSH) did not differ among groups (P greater than .05). Hypophyseal LH was correlated (P less than .01) with body condition score (BCS) at parturition and slaughter (r = .36 and .47, respectively). Prepartum LE diet increased (P less than .05) met-enkephalin in POA compared to prepartum ME (.59 +/- .05 vs. .44 +/- .04 pmol/mg) regardless of postpartum diet or suckling status. Concentrations of beta-endorphin in combined HYP + POA were decreased (P less than .05) by 48 hr CR (15.1 +/- 1.1 vs. 18.1 +/- 0.7 fmol/mg).(ABSTRACT TRUNCATED AT 250 WORDS)

Phylogeny and ontogeny of gonadotropin-releasing hormone: comparison of guinea pig, rat, and a protochordate.

Immunoreactive gonadotropin-releasing hormone (ir-GnRH) was detected in brain extracts of newborn and 10-day-old rats and in adult guinea pigs; it was also present in extracts of the neural ganglion and gland of a protochordate. Radioimmunoassay (RIA) using different GnRH antisera after high-performance liquid chromatography (HPLC) revealed that the dominant form of GnRH is the mammalian form (pGlu-His-Trp-Ser-Tyr-Gly-Leu-Arg-Pro-Gly-NH2) both during ontogenesis in the rat and in the adult guinea pig known to have variant forms of other peptide hormones. None of the other forms of GnRH identified in nonmammalian species to date appear to be present in the rat or guinea pig. A small amount of an unidentified HPLC early eluting form of GnRH is present, but detection by antiserum B-6 implies that it is also mammalian GnRH, with the possibility of changes in positions 2-4. The molecular form of GnRH in a protochordate, the sea squirt Chelyosoma productum, is distinct from salmon and mammalian GnRHs. Cross-reactivity with the sea squirt GnRH-like molecule was highest with an antiserum made against lamprey GnRH; the same antiserum was used to stain nerve fibers in the neural ganglion and some of its roots. This is the first report using RIA, HPLC, and immunocytochemistry to show that protochordates have GnRH-like material. The results suggest that GnRH may have been present at the transition between the invertebrates and vertebrates.

Light and electron microscopic immunocytochemical analysis of antibodies directed against GnRH and its precursor in hypothalamic neurons.

A battery of antibodies directed against different portions of the precursor to gonadotropin-releasing hormone (GnRH), as well as to the mature decapeptide, were characterized immunocytochemically in two ways. Absorption experiments were used to determine the epitope recognized by each antiserum. Electron microscopic immunocytochemistry was then used to define the subcellular organelles that contained reaction product when tissue was incubated with these reagents. These latter observations helped to determine if the antibody recognized the epitope as part of the intact precursor or only after it had been cleaved from parent protein. Our results demonstrate that the GnRH precursor is routed from the rough endoplasmic reticulum through the Golgi apparatus to the secretory vesicles. Furthermore, we show that initial cleavage and processing of the GnRH precursor begin in the cell soma. These antibodies should be useful in the future in determining changes in processing of precursor in animals that differ in endocrine function.

Chicken GnRH II occurs together with mammalian GnRH in a South American species of marsupial (Monodelphis domestica).

Two molecular forms of gonadotropin-releasing hormone (GnRH) were demonstrated in hypothalamic extracts of M. domestica using high performance liquid chromatography and radioimmunoassay with specific GnRH antisera. One form eluted in the same position as synthetic mammalian GnRH and was quantified equally by two mammalian GnRH antisera, while the second form coeluted with synthetic chicken GnRH II and was quantified equally with two chicken GnRH II antisera. The finding of chicken GnRH II in a South American species of marsupial, which has previously been reported in some Australian species of marsupial and in species of Aves, Reptilia, Amphibia, Osteichthyes and Chondrichthyes, supports our hypothesis that this widespread structural variant may represent an early evolved and conserved form of GnRH.

Chromatographic and immunological evidence for mammalian GnRH and chicken GnRH II in eel (Anguilla anguilla) brain and pituitary.

Gonadotropin-releasing hormone (GnRH) peptides in the brain and pituitary of the European eel (Anguilla anguilla) were investigated by reverse phase high performance liquid chromatography (HPLC) and radioimmunoassay with region-specific antisera. Two GnRH molecular forms were demonstrated in brain and pituitary extracts. One form eluted in the same position as synthetic mammalian GnRH on HPLC and was recognized by antibodies directed against the NH2 and COOH termini of mammalian GnRH as well as by antibodies to the middle region. The second form eluted in the same position as synthetic chicken GnRH II and was recognized by specific antibodies to this molecule. Salmon GnRH and chicken GnRH I were not detected. The occurrence of mammalian GnRH in teleost fish suggests that this molecular form is more ancient than was previously suspected and arose earlier than in primitive tetrapods, or that it has arisen in the eel through random mutation of salmon GnRH. The lack of salmon GnRH in the eel brain indicates that this molecular form is not common to all teleost species. The finding in eel brain of chicken GnRH II, which has previously been described in species of Mammalia, Aves, Reptilia, Amphibia, Osteichthyes, and Chondrichthyes, supports our hypothesis that this widespread structural variant may represent an early evolved and conserved form of GnRH.

Topographical relationships between catecholamine- and neuropeptide-containing fibers in the median eminence of the newt, Triturus alpestris. An ultrastructural immunocytochemical study.

Dopaminergic and peptidergic nerve fibers were simultaneously demonstrated with a double-labeling technique at the ultrastructural level. The first antibody, raised against tyrosine hydroxylase, was applied during the preembedding phase and visualized with the peroxidase method. The second antibody, raised against one of the peptides met-enkephalin, somatostatin or gonadotropin-releasing hormone (GnRH), was applied to the ultrathin sections and visualized with gold-labeled goat anti-rabbit IgG. The fibers of both categories were present in the zona externa of the median eminence, frequently contacting the basal lamina of the portal vessels. In addition, topographical relationships between different types of nerve fibers were observed in the perivascular areas, although there were no morphological signs of synaptic specializations. Using serial sections, it could be established that one GnRH-fiber contacted both a dopaminergic fiber and a fiber immunoreactive for met-enkephalin. The observations support earlier physiological data concerning the regulation of the hypothalamo-hypophyseal axis, with special emphasis on the release of neurohormones in the median eminence of the newt.

The gonadotropin-releasing hormone neuronal system of the male Djungarian hamster: distribution from the olfactory tubercle to the medial basal hypothalamus.

The neuroanatomical distribution and morphology of neurons that produce gonadotropin-releasing hormone (GnRH) in the brain of the postpubertal male Djungarian hamster was studied using light microscopic immunocytochemistry. Analysis of every section from the rostral olfactory tubercle to the medial basal hypothalamus indicate 356 +/- 37 immunoreactive GnRH perikarya per brain (mean +/- SE; n = 4 brains). Over 90% of GnRH cell bodies were found in 6 brain regions; the largest number of somata were located in the medial preoptic area followed by the diagonal band, lateral hypothalamus, lateral preoptic area, lateral septum and anterior hypothalamus. Morphologically, two predominant types of GnRH neurons were identified: unipolar GnRH cells with an ovoid soma and only a single distinct process (about 40% of all GnRH neurons), and bipolar cells with a fusiform-shaped perikaryon. Overall and in most brain regions, the ratio of unipolar to bipolar GnRH perikarya was 2:3 or greater. A significant proportion of GnRH neurons had an unusually "thick" process(es) that exited the soma and tapered gradually. GnRH fibers were evident in most sections, forming dense plexuses in the arcuate nucleus-median eminence, the periventricular region of the third ventricle and organum vasculosum of the lamina terminalis. These findings indicate that the Djungarian hamster is similar to other rodent species, especially the white-footed mouse, in the neuroanatomical distribution of GnRH neurons. The present report provides a working atlas for the rostral ventral forebrain of the postpubertal Djungarian hamster.(ABSTRACT TRUNCATED AT 250 WORDS)

Molecular forms of immunoreactive gonadotropin-releasing hormone in hypothalamus and testis of the frog, Rana esculenta.

The hypothalamus and the testis of the frog, Rana esculenta, contain gonadotropin-releasing hormone (Gn-RH)-like peptides which are recognized by an antiserum raised against mammalian Gn-RH. Two molecular forms which coelute with synthetic chicken II and salmon Gn-RH from reverse-phase HPLC were distinguished in the hypothalamus. A single peak coeluting with synthetic chicken II Gn-RH was present in the testis.

A new member of the gonadotropin-releasing hormone family in teleosts: catfish gonadotropin-releasing hormone.

Two forms of immunoreactive gonadotropin-releasing hormone (GnRH) were detected in extracts of brain-pituitary tissue from the African catfish, Clarias gariepinus. Catfish I GnRH eluted first from reverse-phase HPLC and was present in larger amounts compared with catfish II GnRH. Chromatographic and immunological studies with four antisera provide evidence that catfish I GnRH is unique compared with identified GnRHs from mammal, chicken, salmon, and lamprey. Catfish II GnRH elutes in the same position as chicken II GnRH and the forms cannot yet be distinguished. GnRHs extracted from female and male catfish tissue appear to be similar in terms of the number of peaks eluted, elution position, quantity, and cross-reactivity with the antisera. The results of the HPLC and radioimmunoassay studies suggest that catfish I GnRH is likely to be 10 amino acids in length, and have an amide at the C terminus similar to the other family members. In addition, catfish I GnRH is probably different in the 5 to 10 amino acid region compared with mammalian GnRH. Finally, catfish I GnRH is likely to have a lysine or arginine residue as it is the most hydrophilic family member. The lack of the salmon form of GnRH and the presence of a unique GnRH form constitute another example of the considerable evolutionary variation that has occurred in the catfish family compared with other teleosts.

Existence of a GnRH immunoreactive nucleus in the dorsal midbrain tegmentum of the chameleon.

The GnRH system of the chameleon brain was studied at light microscopic and ultrastructural levels by use of an immunohistochemical technique with antibodies directed against salmon gonadotrophin-releasing hormone. Immunoreactive (IR) perikarya were found in the anterior midbrain tegmentum. At this level numerous IR cell bodies were detected around the fasciculus longitudinalis medialis (FLM). The more rostral neurons were observed dorsal to the FLM and progressively tended to be lateral to it along the midline. More caudally, they were found ventral to the FLM. At the electron microscope level, these cells were seen to contain large granular vesicles and to receive numerous synaptic inputs. A prominent pathway was traced from these cell bodies along the medulla oblongata to the spinal cord. A second IR pathway ascended rostrally to the habenular complex. No IR perikarya were located in the anterior brain including the olfactory bulbs.

Development of a radioimmunoassay for measuring gonadotrophin releasing hormone in patients receiving treatment.

A radioimmunoassay for the measurement of gonadotrophin releasing hormone (GnRH) in plasma and urine using readily available reagents was developed. The GnRH assay showed good precision, recovery, and parallelism over a wide range of GnRH concentrations with a sensitivity of 15 pg/ml. The assay was compared with a commercially available kit (Buhlmann Laboratories). Although the Buhlmann kit showed acceptable precision, recovery, sensitivity, and correlation with the developed GnRH assay for plasma samples, lack of parallelism of serially diluted plasma and urine samples was consistently observed, together with a poor correlation with the developed GnRH assay for urine, suggesting a matrix effect with the Buhlmann kit. The developed assay is suitable for measuring GnRH in samples obtained from patients receiving pulsatile infusions of GnRH. In contrast, the commercially available Buhlmann kit was unsuitable for measuring plasma GnRH as the kit had a top standard of only 160 pg/ml, well below the peak plasma concentration. It would not be possible to dilute samples for analysis because of the lack of parallelism of diluted samples compared with standards obtained with the Buhlmann assay.

Bromocriptina na síndrome dos ovários policísticos: uma revisäo / Bromocriptine in polycystuc ovary syndrome: a review

A síndrome dos ovários policísticos (SOP) é uma entidade cuja fisiopatologia näo está totalmente esclarecida e os tratamentos propostos apresentam resultados variáveis e insatisfatórios. A possibilidade de que alteraçöes dopaminérgicas hipotalâmicas ocasionassem um distúrbio na secreçäo do hormônio liberador das gonadotrofinas (GnRH) e, em conseqüência, um aumento exagerado do hormônio luteinizante (LH), levou autores a utilizar a bromocriptina (BRC), um potente agonista da dopamina, no tratamento desta síndrome. No entranto, os resultados têm sido confiltrantes, havendo que descrevem sucesso de seu uso e outros que näo conseguiram demonstrar sua utitlidade. Nós revisamos a literatura disponível sobre o controle da secreçäo do LH pela dopamina (DA), as discrepância existentes com o uso da BRC na SOP e os mecanismos de açäo que podem estar envolvidos. Concluímos que a utilidade da BRC na SOP bem como seu mecanismo de açäo, necessitam de trabalhos adicionais para melhor esclarecimento