Behaviour of a somatotroph population under a growth hormone releasing peptide treatment.

In this investigation, we studied the effects of Momany peptide (GHRP-5), on somatotroph secretory activity. Acute and chronic administration of GHRP-5 provokes a significant release of growth hormone that can be closely correlated with ultrastructural changes in somatotroph populations. After 3,5 and 7 days of GHRP-5 treatment, two somatotroph cell subpopulations coexist. One of them has an enhanced secretory activity and the other presents a quiescent appearance. Therefore, pituitary growth hormone content was not affected in the first seven days of GHRP-5 treatment. After 14 days, there was a significant depletion of growth hormone pituitary content coincident with the highest levels of serum growth hormone. These results concur with the surge of a new hyperactive somatotroph subtype characterised by numerous immature secretory granules that are discharged bypassing the maturation step. Acute and chronic treatments caused no changes in somatotroph cell density, the area immunostained for growth hormone and the levels of total mRNA for transcription factor pit-1. The results of pituitary cell cultures incubated with specific blockers for different signalling pathways demonstrated an involvement of the phospholipase C-inositol phosphate system in GHRP-5 stimulated somatotroph secretion. GHRP-5 treatment enhanced significantly the release of growth hormone, thereby eliciting ultrastructural modifications in somatotrophs that can be correlated with an increased secretory activity devoid of cell density changes.

Uteroglobina endometrial en pacientes con serología positiva para chlamydia trachomatis / Endometrial uteroglobin in patients with positive serology for chlamydia trachomatis

Introducción: Chlamydia trachomatis es causa frecuente de enfermedad inflamatoria pélvica y esterilidad tubaria. La afección endometrial asociada a este germen es dificil de poner de manifiesto. La úteroglobina (Utgh) es una proteína de secreción del tracto respiratorio y genitourinario. En el endometrio se ha detectado durante el comienzo y mitad de la fase selectora. Nuestro objetivo fue localizar la Utgb endometrial por inmunohistoquímica, en pacientes que consultaron por esterilidad y presentaron serología positiva contra C. trachomatis. Material y Metodos: Se estudiaron 30 parejas a las que se les realizó: análisis de esperma, dosajes hormonales, histerosalpingografía, anticuerpos séricos contra C.trachomatis (IgG Cht) y biopsia endometrial (días 19-21 del ciclo). En las biopsias se determinó fechado y presencia de endometritis. La inmunohistoquímica para Utgb se realizó en cortes de parafina usando antisuero policlonal (1:50 y 1:300). Resultados: De las muestras evaluadas, 20 presentaron fechado endometrial acorde al día del prelevamiento (día 19-21). La inmunomarcación para Utgb, fue positiva en el 100 por ciento (20/20) de los casos, con la dilución de anticuerpo 1:50. Se encontró título de IgG Ch t incrementando en 11 pacientes (11/20, 55 por ciento). El 91 por ciento (10/11) de estas presentaron inmunomarcación para Utgb, mientras que el grupo con serología negativa, se detectó en el 44,5 por ciento (4/9) cuando se empleó la dilución 1:300 (p<0.02). La incidencia de endometritis y los niveles de progesterona sérica no mostraron diferencias significativas entre estos grupos. Conclusión: Hemos observado una mayor incidencia de afección endometrial en pacientes con serología positiva para C trachomatis. La inmunodetección de uteroglobina podría ser un marcador de dicha afección, que complementaría la información obtenida por microscopía óptica (AU)

Uteroglobina endometrial en pacientes con serología positiva para chlamydia trachomatis / Endometrial uteroglobin in patients with positive serology for chlamydia trachomatis

Introducción: Chlamydia trachomatis es causa frecuente de enfermedad inflamatoria pélvica y esterilidad tubaria. La afección endometrial asociada a este germen es dificil de poner de manifiesto. La úteroglobina (Utgh) es una proteína de secreción del tracto respiratorio y genitourinario. En el endometrio se ha detectado durante el comienzo y mitad de la fase selectora. Nuestro objetivo fue localizar la Utgb endometrial por inmunohistoquímica, en pacientes que consultaron por esterilidad y presentaron serología positiva contra C. trachomatis. Material y Metodos: Se estudiaron 30 parejas a las que se les realizó: análisis de esperma, dosajes hormonales, histerosalpingografía, anticuerpos séricos contra C.trachomatis (IgG Cht) y biopsia endometrial (días 19-21 del ciclo). En las biopsias se determinó fechado y presencia de endometritis. La inmunohistoquímica para Utgb se realizó en cortes de parafina usando antisuero policlonal (1:50 y 1:300). Resultados: De las muestras evaluadas, 20 presentaron fechado endometrial acorde al día del prelevamiento (día 19-21). La inmunomarcación para Utgb, fue positiva en el 100 por ciento (20/20) de los casos, con la dilución de anticuerpo 1:50. Se encontró título de IgG Ch t incrementando en 11 pacientes (11/20, 55 por ciento). El 91 por ciento (10/11) de estas presentaron inmunomarcación para Utgb, mientras que el grupo con serología negativa, se detectó en el 44,5 por ciento (4/9) cuando se empleó la dilución 1:300 (p<0.02). La incidencia de endometritis y los niveles de progesterona sérica no mostraron diferencias significativas entre estos grupos. Conclusión: Hemos observado una mayor incidencia de afección endometrial en pacientes con serología positiva para C trachomatis. La inmunodetección de uteroglobina podría ser un marcador de dicha afección, que complementaría la información obtenida por microscopía óptica

Somatotroph response to periodical IGF-I administration to male rats.

Insulin-like growth factor I (IGF-I) downregulates growth hormone (GH) expression in pituitary cell cultures. However, in vivo different results were found depending on the experimental protocol used. We determined the kinetics of changes of pituitary and serum GH concentrations after subcutaneous IGF-I administration (240 microg/100 g body weight) to rats every 12 h for various periods. These parameters were correlated with changes in the somatotroph cell population. A significant increase in serum GH was registered at 6 h after IGF-I injection. At this time, some somatotroph cells exhibited ultrastructurally signs of high secretory activity, whereas adjacent somatotroph cells showed a quiescent appearance with sizeable stores of secretory granules. In contrast, serum GH levels remained unchanged at 1, 2 and 12 h after each IGF-I injection. Pituitary GH concentrations were comparable to control levels during the first 48 h and declined significantly at 72 h and 96 h of IGF-I treatment. After these prolonged periods of time of treatment, the size and extension of organelles involved in protein synthesis decreased and mature secretory granules in the cytoplasm increased significantly in GH-secreting cells. The somatotroph cell density remained unchanged even at 96 h of treatment. In conclusion, our results suggest that periodical IGF-I administration to rats does not inhibit GH secretion. Interestingly, IGF-I injections induced early and significant increases in serum GH levels. This result may be a consequence of a temporary stimulatory action on somatotroph cells concurrent with increased secretory activity.

Cellular and functional interactions between gonadotrophs and lactotrophs in pituitary cell cultures.

Lactotroph secretory activity is regulated by hypothalamic stimulating and inhibiting factors as well as peripheral endocrine hormones. In addition to this important control domain, the pituitary gland displays an intrinsic regulatory ability through autocrine and paracrine signals. To evaluate the role of gonadotrophs in the regulation of prolactin (PRL) secretion, a comparative study was performed applying two regulatory agents that operate through different physiological mechanisms: gonadotropin-releasing hormone (GnRH), which releases regulatory factors co-localized in secretory granules of gonadotrophs, stimulating PRL secretion from lactotrophs; and angiotensin II (AII), with direct effects on lactotroph secretion through specific receptors. In these studies performed in regular and purified primary pituitary cell cultures from female rats, the lactotrophs comprised the largest population of cells (about 51%), whereas gonadotrophs represented only a small fraction (3%) of the total pituitary cell count. In regular cell cultures treatments with AII and GnRH showed a similar secretory behavior, increasing PRL output 73% and 63%, respectively. The stimulation with GnRH and AII of cell cultures with purified lactotrophs and gonadotrophs provided comparable results, but the response of lactotrophs was significantly higher (106% and 138%, respectively) than that recorded in regular cell cultures. Simultaneous AII treatment with an antipeptide antagonist to AII receptor (AII-antipep) completely blocked the PRL release induced by AII. The co-incubation of cells with GnRH and AII-antipep suppressed the peak of PRL release caused by GnRH, confirming that AII is a paracrine agent released by gonadotrophs stimulated with GnRH. The different secretory behavior of lactotrophs treated with AII and GnRH in both regular and purified cell cultures is indicative of the degree of functional interactions between different pituitary cell types. The present study supplies morphological and functional information on the cell-to-cell interactions, which plays an important role in the intrinsic regulatory control of PRL secretion.

Biochemical and functional changes during the bovine fetal thyroid development.

To establish biochemical and functional relations during thyroid development, the activity of thyroid peroxidase (TPO), nicotinamide adenine dinucleotide phosphate (NADPH)-cytochrome c reductase and monoamine oxidase (MAO) in a particulate fraction and the iodide transport and organification in slices of bovine fetal thyroid were examined throughout gestation. The cytochemical localization of TPO, H2O2 generating sites and MAO was also studied. Fetal glands were grouped in stages I to V according to increasing developmental features; adult tissues were also analyzed. TPO activity in each of the fetal stages was higher than in the adult; a marked increase was observed in stages IV and V. Iodide transport (T/M) was similar in stages I to V and the adult. Iodide organification in fetal thyroids showed a similar pattern to that of TPO activity. When compared with the adult, at midgestation (stages II to III), a lower iodination coexisted with a higher TPO activity. The activity of NADPH-cytochrome c reductase and MAO, two enzymes previously proposed to participate in thyroid H2O2 generation, did not parallel the level of iodide organification. Cells from stages II to V exhibited a positive cytochemical reaction for TPO in the rough endoplasmic reticulum (RER) and the perinuclear cisternae (PC). In stages IV, V, and adult, TPO was occasionally found in apical vesicles and microvilli, whereas H2O2 was detected within the RER and the PC. MAO reaction was positive in adult, but not in fetal thyroid. These results indicate that a high TPO activity accompanied the onset of the organification process during fetal thyroid development. The level of iodination was associated with the presence of TPO at a proper site rather than to the level of TPO activity. Evidence against a role of NADPH-cytochrome c reductase and MAO in the iodide organification was obtained.

Residual effects of thyroid hormone on secretory activity of somatotroph population.

In the present experiments the effects of the interruption of a prolonged T3 treatment on somatotroph population and GH synthesis and secretion were studied. The treatment with pharmacological doses of T3 provokes marked ultrastructural changes in somatotrophs compatible with a stimulated synthesis of GH. These results can be correlated with the significant increase in pituitary GH content and the normal values of serum GH. Twenty-four hours after T3 withdrawal, somatotrophs exhibited a marked depletion of secretory granules by exocytosis. These changes were concurrent with a significant discharge of pituitary GH and a two-fold increase in GH serum levels. The serum concentrations attained the highest values on the second and the third day after the T3 suppression, while the pituitary GH contents recovered the control levels. Morphometry of somatotroph population revealed a clear proliferation of cells and increased areas immunostained for GH, 24 h after withdrawal of the T3 treatment. The effects of T3 on somatotrophs were persistent for several days and at least five days were required for all parameters to return to control values. These results provide a new insight on the residual activity of thyroid hormones on both functional activity and morphological organization of somatotrophs.

Influence of L-triiodothyronine on rat somatotroph cells.

The L-Triiodothyronine (L-T3) has a direct influence on the population of somatotrophs in rat pituitary gland. This effect is dose-dependent and induces both proliferation of somatotrophs and striking changes in the synthesis and secretion of growth hormone (GH). Daily injections of 5 micrograms L-T3 for 7 days increased significantly the synthesis and storage of GH in pituitary gland, but the GH release was partially blocked. By contrast, injections of 10 micrograms L-T3 promote rapid synthesis and secretion of GH with removal of the cytoplasmic stores of the hormone and a consequent rise of serum levels. A close correlation was found between levels of stimulation and proliferation or retrogression of lactotroph cell population.

Dissociation of thyrotropin-dependent enzyme activities, reduced iodide transport, and preserved iodide organification in nonfunctioning thyroid adenoma and multinodular goiter.

Several biochemical and functional modifications demonstrated in goitrous tissues could reflect the effect of goitrogenic factors. Growth-enhancing agents, including TSH itself, have been involved in goitrogenesis. To study comparatively the variation patterns of some TSH-dependent enzymes within single goitrous tissues, we measured the activities of peroxidase (TPO), NADPH-cytochrome-c (cyt-c) reductase, and monoamine oxidase (MAO) in tissues from cold follicular adenoma and multinodular goiter. Iodide transport and organification were also evaluated. Perinodular and necropsy tissues were used as controls. The mean TPO activity measured by guaiacol as well as triiodide assays was significantly increased in multinodular goiter, whereas a nonsignificant increment was observed in cold adenoma. NADPH-cyt-c reductase and MAO were markedly increased in the two types of pathological tissues. The individual activities of the three enzymes showed dissimilar modifications within single samples and among different tissues. There was no correlation in the activities of the enzymes within single specimens from cold adenoma and multinodular goiter, except for MAO and NADPH-cyt-c reductase in multinodular goiter, for which a significant correlation was obtained. In this tissue, MAO and TPO measured by guaiacol assay were weakly correlated. TPO activity evaluated by guaiacol oxidation was correlated with that measured by triiodide formation in cold adenoma, but not in multinodular goiter. The mean iodide organification values assayed by iodotyrosine formation in the absence of exogenous H2O2 in particulate fractions from cold adenoma and multinodular goiter were within the normal range. A reduced iodide transport, evaluated as the thyroid/medium ratio, was observed in slices from these tissues. The dissociation of the three enzyme activities in single specimens from cold adenoma and multinodular goiter along with the reduced iodide transport in these tissues support the hypothesis that factors other than TSH or with TSH-like effects could be involved in the abnormal thyroid growth.

Sustancias complejantes que facilitan la producción de clamidosporos para la identificación de Cándida albicans / Complex substances that facilited the production the clamidospores for the identification of Candida albicans

La influencia de polisacáridos y de ciertas condiciones físico-químicas es conocida en la formación de seudomicelios y clamidosporos, como las producidas por la presencia de sustancias tensioactivas. Se procuró establecer, hasta qué punto sustancias complejantes como el EDTA (sal sódica del ácido etilendiaminotetracético), el citrato de sodio y el oxalato de sodio, podrían igualmente influir en el dimorfismo del hongo. Se utilizaron cepas de Candida albicans aisladas de procesos patológicos e identificadas por zimograma y auxanograma. Fueron sembradas en medios de agua harina de maíz y agua papa-zanahoria, a los que se les había incorporado los complejantes por separado. Como controles se emplearon los mismos medios sin aditamentos con el agregado de tween 80. Las concentraciones se fueron regulando por estudios previos hasta poder obtener las más útiles para nuestros fines. Se comprobaron buenos resultados con la incorporación de los distintos complejantes para la formación de filamentos y clamidosporos, pero fueron inferiores a los resultados obtenidos con el uso del tensioactivo (tween 80) (AU)

Sustancias complejantes que facilitan la producción de clamidosporos para la identificación de Cándida albicans / Complex substances that facilited the production the clamidospores for the identification of Candida albicans

La influencia de polisacáridos y de ciertas condiciones físico-químicas es conocida en la formación de seudomicelios y clamidosporos, como las producidas por la presencia de sustancias tensioactivas. Se procuró establecer, hasta qué punto sustancias complejantes como el EDTA (sal sódica del ácido etilendiaminotetracético), el citrato de sodio y el oxalato de sodio, podrían igualmente influir en el dimorfismo del hongo. Se utilizaron cepas de Candida albicans aisladas de procesos patológicos e identificadas por zimograma y auxanograma. Fueron sembradas en medios de agua harina de maíz y agua papa-zanahoria, a los que se les había incorporado los complejantes por separado. Como controles se emplearon los mismos medios sin aditamentos con el agregado de tween 80. Las concentraciones se fueron regulando por estudios previos hasta poder obtener las más útiles para nuestros fines. Se comprobaron buenos resultados con la incorporación de los distintos complejantes para la formación de filamentos y clamidosporos, pero fueron inferiores a los resultados obtenidos con el uso del tensioactivo (tween 80)