Search PubMed⌕ Search

Biomedical subjects

A Rosado

Publications and source records attributed to A Rosado.

At least 37 records · Page 2Linked to original sources
PubMed1997

Proline-glutamate interactions in the CNS.

1. Crude synaptosomes (P2) and synaptosomal membranes were prepared from normal C57/B110 mouse brains and Wistar rats respectively. 2. [3H]Pro binding to mouse brain synaptic membranes was examined in the presence of competitive NMDA antagonist, MK-801, or HA-966. Conversely, the effects of l-proline on [3H]MK-801 binding were also probed. The effects of l-proline on glutamate-medicated [Ca+2]i levels were tested. 3. The authors could not detect any effect of proline on glutamate-mediated [CA+2]i levels using FURA-2 in synaptosomes or neuroblastoma cells. 4. NMDA competitive antagonists, AP-7, CPP, and CGS 19755 inhibit [3H]Pro binding to mouse brain synaptic membranes. 5. MK-801, a NMDA channel blocker, also inhibits [3H]Pro binding, but 200 mM proline is incapable of inhibiting [3H]MK-801 binding. 6. HA-966, a glycine site partial agonist inhibits [3H]Pro binding. Proline has modest effects on [3H]glycine binding.

Animals↗
PubMed1997

Metalloproteinase activity during growth, maturation and atresia in the ovarian follicles of the goat.

Metalloproteinases are an important group of hydrolytic enzymes which participate in interstitial matrix degradation during tissue remodelling processes and therefore may be required during follicular growth and maturation. The activity of metalloproteinases (collagenases, gelatinase, and Pz-peptidase), was measured during growth, maturation and atresia of goat antral follicles. These follicles (n = 67) were separated by size and also classified into four groups: non-atretic (Group I); early atretic (Stage I) (Group II); moderately atretic (Stage II) (Group IIIa); and, late atretic (Stage III) (Group IIIb). Pz-peptidase was greater in granulosa than in thecal cells, and almost absent in follicular fluid. In non-atretic follicles, activity in granulosa cells increased with increasing follicle size, whereas activity peaked in 3-6 mm follicles in thecal cells. Atresia was associated with declining activity in thecal cells from follicles in the 3-6 mm range and in granulosa cells from the > 6 mm range. Interstitial collagenase activity was significant and similar in granulosa and thecal cell extracts and low in follicular fluid from non-atretic follicles. Activity increased significantly in thecal cells, but decreased significantly in granulosa cells from large (> 6 mm) non-atretic follicles. Atresia was associated with declining activity in both types cells and increasing activity in follicular fluid. Gelatinase activity was some times associated with five regions corresponding to molecular weights of 22.1, 30.7, 39.6, 63.8 and 71.4 kDa, and rarely at 91.3 and 81.2 kDa. Overall activity declined with atresia in thecal cells from follicles in the 3-6 mm range, but not in those > 6 mm. In granulosa cells from follicles 3-6 mm, activity varied widely with stage of atresia, while in cells from follicles > 6 mm, activity was greatly increased in atretic follicles.

Animals↗
PubMed1997

Deconvolution analysis of bioassayable LH secretion and half-life in men with idiopathic oligoasthenospermia.

To further investigate the nature of neuroendocrine disturbances of the hypothalamopituitary-gonadal axis in idiopathic male infertility, we studied 12 infertile men with oligoasthenozoospermia and 13 euspermic controls, matched for age and body mass index, by blood withdrawal at 10-min intervals for 8 h to analyse pulsatile release of bioactive LH (b-LH). The rat interstitial cell testosterone (RICT) bioassay was used in conjunction with a recently validated multiparameter deconvolution algorithm, to estimate the endogenous half-life of b-LH, its secretory burst frequency, amplitude, duration and mass. Oligoasthenospermic men exhibited significant (p < 0.05) alterations within the LH axis; namely: (1) a prolonged half-life of b-LH (92 min in euspermic men, 127 min in oligoasthenospermic men); (2) a reduced b-LH secretory burst amplitude (2.2 +/- 1.2 IU/l/min in euspermic men, 1.7 +/- 0.8 IU/l/min in oligoasthenospermic men); (3) a lower bioactive/immunoactive (b/i) ratio for LH secretory burst amplitude (14 in euspermic men, 4 in oligoasthenospermic men); (4) a reduced b/i ratio in the mass of LH secreted per burst (5.4 in euspermic men, 4.1 in oligoasthenospermic men) and (5) decreased coordinate release of b-LH and testosterone in infertile men, as assessed by cross-correlation analysis. These disturbances differ from the neuroendocrine dysregulation described in other states of male hypogonadotrophism.

Activity Cycles↗
PubMed1997

[The male factor. I. Testicular function and gametogenesis].

The knowledge and treatment of human reproduction impairments have had an outstanding development during the last 15 years, mainly in topics related with infertility and sterility. Unfortunately in the area of male reproduction development has not been the same and despite the efforts accomplished, progress in the diagnosis and therapeutics of male infertility is limited. The functions of the male gonad are sperm production as well as the synthesis and secretion of steroid hormones, mainly testosterone. The efficient regulation of these functions depends on both the precise pituitary secretion of gonadotropic hormones and the endocrine, paracrine and autocrine responses of the testicular somatic cells to this stimulation. In this review we discuss the main events associated with the spermatogenic process, including the interactions among the testicular somatic cells and some characteristics of the physiological function of the hypothalamus-pituitary-testicle axis, as well as some pathophysiological states related to male infertility due to "pretesticular causes".

Gametogenesis↗
PubMed1997

[The male factor. II. Spermatozoa. Structure and function].

In the "male factor" entity, the structural and functional correlation determining the sperm fertilizing capacity is constituted by a group of cellular factors that must be evaluated in the semen of the infertile men. Structurally the spermatozoa of the head, the middle piece and the flagellum. The head has a highly condensed haploid nucleus, surrounded by a thin layer of cytoplasmic material, which is covered in a cap-like fashion by the membrane limiting the acrosome. This last organelle, which has characteristics similar to those of a secretory granule, secretes in a programmed way the hydrolytic enzymes that facilitate the fertilization process. The middle piece contains the mitocondrail sheet, responsible for the energy metabolism of the sperm cell. The flagellum has the same basic structure of other cilia or flagella, but also has particular characteristics due to the presence of the outer dense fibers and the fibrous sheath. In the semen analysis from infertile men the abnormalities most frequently observed belong to the number, morphology, variability, motility of capacitation-acrosome reaction of the sperm cells. However, due to the apparent multifactorial etiology of the male factor, now a days we only have few options for medical or pharmacological treatment. In this paper we review the morphology and ultrastructure considered as "normal" in the human spermatozoa, as well as we describe the most frequent alterations in these parameters. At the same time, we discuss the impact of sperm motility and of the capacitation-acrosome reaction process in male fertility.

Acrosome↗
PubMed1997

[The male factor. III. Importance, diagnosis and perspectives].

According to the World Health Organization, the male factor is present in 40-50% of infertile couples, therefore the importance of the continuity in multidisciplinary studies related to understand the mechanisms regulation the male reproductive process. Semen analysis is still almost the only instrument for the study of men with disruptions in their fertility. However, the main problem to establish a diagnosis with predictive value for male infertility, is the fact that the morphological and/or functional characteristics of the male gamete that determine its fertilizing capacity are still in the process of validation. Even more, the correlations among semen characteristics and bioactive hormones concentrations in serum, and the individual fertility potential have not been established either. In this context, we discuss here some topics that we considered of great importance in relation to male fertility diagnosis, i.e. the main causes of infertility, the advances in the recognition of the morpho-functional correlation that determines the fertilizing capacity of the male gamete and the criteria that prevail in the laboratory to proper handle and evaluate the semen samples.

Fertility↗
PubMed1996

Ca2+/calmodulin system: participation in the progesterone-induced facilitation of lordosis behavior in the ovariectomized estrogen-primed rat.

We studied the effect some drugs that participate in the Ca2+/ calmodulin system have on the progesterone (P) facilitation of lordosis behavior in ovariectomized estradiol (E2) primed rats. We injected rats 44 h after E2 priming with 2 mg P together with various dosages of one of the following compounds: pentobarbital, trifluoperazine (TPZ), promethazine (PMZ), Chlorpromazine (CPZ), haloperidol (HAL), pimozide (PIM), and verapamil (VER). Then 4 h after treatment, animals were tested for sexual behavior, expressed as the lordosis quotient (LQ). All drugs at 4 mg/kg or higher inhibited lordosis, but only HAL, PIM, and VER were active at 1 mg/kg. The maximum level of activity was shown by PIM, although at the dose of 8 mg/kg no statistical differences were found between this compound and TPZ or HAL. Pentobarbital (25 mg/kg) showed no significant difference from saline-treated controls. The activity of the tested drugs on the facilitation of sexual behavior appears to be related to their efficiency as inhibitors of calmodulin (CaM)-dependent phosphodiesterase and as ligands for the Ca(2+)-CaM complex.

Animals↗
PubMed1996

Alterations in pulsatile luteinizing hormone and follicle-stimulating hormone secretion in idiopathic oligoasthenospermic men: assessment by deconvolution analysis--a clinical research center study.

To investigate the nature of neuroendocrine disturbances of the hypothalamo-pituitary-gonadal axis in idiopathic male infertility, we studied 14 infertile men with oligoasthenozoospermia (OLIGO) and 15 age-, body mass index-, and community-matched euspermic controls by blood withdrawal at 10-min intervals for 12 h to encompass basal (8-h) and exogenous GnRH-stimulated (4-h) pulsatile release of LH and FSH (by immunoradiometric assay) as well as testosterone (by RIA). Deconvolution analysis was used to estimate endogenous LH and FSH half-lives, secretory burst frequency, amplitude, duration, and mass. OLIGO men exhibited normal serum concentrations of total, free, and percent dialyzable testosterone and estradiol, but distinct dynamic alterations within the LH and FSH axes; namely (P < 0.05), 1) a prolonged half-life of LH (OLIGO, 95 +/- 19 min; control, 80 +/- 9.3 min) and a reduced half-life of FSH (OLIGO, 260 +/- 79 min; control, 320 +/- 93 min); 2) a low LH, but normal FSH, secretory burst frequency (OLIGO, 12 +/- 3.4; control, 15 +/- 3.0 LH pulses/day); 3) a decreased serum testosterone peak frequency (OLIGO, 16 +/- 4.3; control, 21 +/- 3.2 peaks/day); and 4) an amplified mass of LH (1.1- to 1.3-fold higher in OLIGO) and FSH (2.4- to 2.7-fold higher in OLIGO) secreted per burst basally as well as after GnRH injection. These disturbances were readily distinguishable from the neuroendocrine dysregulation described in other states of male hypogonadotropism (e.g. uremia, fasting, and aging).

Adult↗
PubMed1996

Changes in lysosomal enzyme activities in pre-ovulatory follicles and endometrium of PMSG superovulated rats.

Large superovulatory doses of gonadotrophins result in reduced fertility in laboratory and large domestic animals and it has been postulated that some of the superovulated oocytes are derived from abnormal follicles which would not ovulate under normal physiological stimuli. Follicular growth, follicular maturation and atresia, ovulation and the nidation of the fertilized oocyte require intense tissue remodelation which can be accomplished only through the action of hydrolytic enzymes. We have studied the activities and sub-cellular distribution of three lysosomal enzymes (acid phosphatase, N-acetyl-beta-D-glucosaminidase and beta-glucuronidase) in the follicular fluid, granulosa and theca cells of preovulatory follicles and in the endometrial tissue of immature Wistar rats injected with 4 (control) or 40 (superovulated) IU of pregnant mare serum gonadotrophin (PMSG). Enzyme activities were from four to ten times higher in theca than in granulosa cells. This difference was particularly important in the case of beta-glucuronidase. Large preovulatory follicles tended to have higher activities of lysosomal enzymes in the free fraction of all the compartments studied. This difference was remarkable in theca cells where free enzymes would be required to help ovulation. Forty IU of PMSG induced higher activities of acid phosphatase in theca and granulosa cells than 4 IU, but in endometrial tissue this latter dose of PMSG was more efficient to induce higher activities of this enzyme. The endometrial bound fraction of N-acetyl-beta-D-glucosaminidase was almost three times higher than the free activity. This behavior was also observed in endometrial beta-glucuronidase but only in the control rats. The results observed in follicular fluid were less homogeneous. The activities of glucosaminidase and acid phosphatase were two to three times higher in rats overstimulated with 40 IU of PMSG than in the control rats, whereas the activities of beta-glucuronidase were lower in the superovulated rats. Our results suggest that alterations in the process of tissue remodeling required for ovulation of mature, normal oocytes and for nidation of the fertilized ovum may be important factors to explain pregnancy failure in the PMSG superovulated female.

Animals↗
PubMed1995

[Kinetic study of pulsatility of gonadotropin and testosterone secretion in normospermic fertile and infertile men with idiopathic oligo-asthenozoospermia].

Idiopathic oligo-asthenozoospermia is among the most frequent causes of male infertility and has a not very well understood etiopathogenesis. To obtain valuable information about the role of some endocrine factors in the etiology of this kind of infertility, information that is not easy obtain by the traditional analytical methods, we applied some recently proposed mathematical algorithms to analyze with more exactitude the importance of the secretory pulses of three hormones, luteinizing hormone (LH), follicle stimulating hormone (FSH) and testosterone (T). Serum samples were obtained every 10 min for 12 h, from 15 fertile normospermic men and 14 infertile patients with idiopathic oligo-asthenozoospermia; the concentration profiles of FSH and T were analyzed by IRMA and RIA, and the immuno-and bioactive LH concentrations were quantified by IRMA and bioassay (JCEM 42:958, 1976). To assess the pituitary stores of LH and FSH, after 8 h of spontaneous secretion we administered (2 h apart) 2 intravenous pulses containing 10 micrograms of a GnRH analog, and the sampling continued as described. Hormonal pulsatility was assessed by a computerized cluster analysis method (Am J Phys 250:E486, 1986) an by the multiple parameters deconvolution method (JCEM 66: 1291, 1988). In the infertile patients we found a significant diminution in the length and frequency of LH pulses, compared with the normospermic men. However, LH half life, the interpulse interval, the amplitude and the mass secreted per pulse rose in the infertile males compared with the controls. The increase in the LH half life suggests the secretion of a more acidic isoform of this hormone in the infertile group. After the GnRH injection the LH secreted mass and mean concentration rose significantly in both groups; this effect was higher in the infertile oligo-asthenozoospermic men. In this group we also found a decrease in the bioactive LH interpulse interval and therefore more pulses during the sampling interval, that produced a higher concentration of this kind of hormone in these patients. Oligo-asthenozoospermic men secreted approximately 70% more bioactive LH as a response to the first GnRH injection than the normal controls. The desensitization observed with immunoactive LH (diminution in the mass secreted after the second GnRH bolus compared with the first one) was also observed with bioactive LH. In the infertile men group we found a significant reduction in the FSH half life compared with the normospermic controls; this fact suggests that, contrarily to the results observed with LH, a more basic isoform is secreted in these patients.(ABSTRACT TRUNCATED AT 400 WORDS)

Adult↗
PubMed1993

Ca2+/calmodulin system: participation on rat sexual hypothalamic differentiation.

Modifications of male rat hypothalamic sexual differentiation after neonatal administration of drugs that participate on the Ca2+/calmodulin system (haloperidol, trifluoperazine, penfluridol, pimozide, and verapamil) were studied. Pups treated 72 h after birth were behaviorally tested on day 120 of extrauterine life. Five tests for homotypical behavior were conducted. Afterwards animals were castrated and tested twice for heterotypical (female) behavior under replacement hormonal therapy. Fifty percent (80% in the case of pimozide) of all treated males showed lordotic behavior compared with none of the controls. Haloperidol (39%, lordosis quotient) and pimozide (40%, lordosis quotient) were more active than the others. Results obtained with verapamil were not statistically different from the controls. Pimozide was the most active agent influencing the appetitive masculine behavior (mount latency, intromission latency, and postejaculatory interval). Verapamil was more efficient than the rest of the drugs on the consummatory behavior (mount latency, intromission frequency, interintromission interval, and ejaculatory latency). Our results support the participation of the Ca2+/calmodulin system in hypothalamic sexual differentiation and in the differential modulation of the masculine and feminine behavioral patterns.

Animals↗
PubMed1992

RNA metabolism during the sexual differentiation of rat hypothalamus.

Modifications in the basal molecular biology parameters (total concentrations of RNA, DNA, proteins, rRNA, tRNa, free and polysomal bound poly-A+ mRNA) have been determined daily in the growing hypothalamus of male and female rats from day 1 to day 8 after birth. Changes observed in the parameters studied in this work occurred mainly in the first 48 h after birth. In males tRNA and free mRNA (f-mRNA) contents decreased from day 1 to day 2 and then their concentrations remained more or less constant. Total mRNA significantly decreased from day 1 to day 3 and showed a further significant decrease from day 6 to day 8. Polysomal bound-mRNA (b-mRNA) decreased from day 1 to day 3, then increased to day 6, and finally decreased once more from day 6 to day 8. The b-mRNA/f-mRNA, mRNA/rRNA and mRNA/total RNA ratios showed a bimodal behavior with a first peak on day 2, and a second, smaller peak, on days 6-7. The changes observed in the females on the first 2-3 days of life were the inverse of those observed in the males, most of the parameters studied showed a sharp increase from day 1 to day 2 or to day 3 and then a drastic decrease. The only exception to this behavior was the b-mRNA/f-mRNA ratio which showed a small decrease from day 1 to day 2, followed by a continuous increase from day 2 to day 8. b-mRNA concentrations, after the sharp decrease from day 2 to day 3 of life, increased from day 3 to day 7.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1992

Capacitation and acrosome reaction of human spermatozoa. An assisted reproduction approach.

In this paper we briefly review the recent advances in the understanding of human sperm capacitation, fundamentally focused to give help to the practitioners involved in assisted reproduction. Sperm capacitation has been the subject of study for many years in several animal species, but during the past ten years the advances in assisted reproduction procedures have made important contributions in the progress of this topic in relation to human spermatozoa. In this manuscript we analyze the metabolic, membranal and physiological changes observed in these cells during in vitro capacitation and their relationships with the female genital tract secretions, as well as with the homologous oocytes.

Acrosome↗
PubMed1990

Differences in lipoprotein composition between heads and tails of human sperm: an infrared spectroscopy study.

Human spermatozoa and their fractions (heads and tails) have been studied by infrared spectroscopy. Protein conformation in isolated human spermatozoa heads, although predominantly of the alpha helix or random coil type, has a significant proportion of antiparallel B structure. Spectra of isolated spermatozoa tails show that proteins exist in this fraction preponderantly in pleated-sheet conformation (parallel and antiparallel). The quantity and type of lipids seem to be drastically different between heads and tails of spermatozoa. Head lipids are scarce and difficult to extract, and they are apparently tightly bound to proteins, highly unsaturated, and rich in free hydroxyl and carboxyl groups. Tail lipids are more abundant and more easily extractable. Head phospholipids are probably phosphatidylcholine, cephalins, and inositols, and tail phospholipids are preponderantly plasmalogen-type lecithins and sphingomyelins. The presence of specific infrared bands points to the existence in tails of important amounts of sulfur compounds, probably sulfolipids or sulfoglycolipids.

Humans↗
PubMed1990

Glycosaminoglycan-sulfate as plasma membrane component of pig spermatozoa.

The effect of specific glycosaminoglycan-hydrolyzing enzymes on the ruthenium red staining of pig spermatozoa was studied. Washed spermatozoa were incubated at 35 degrees C in buffer or with neuraminidase 0.5 units/ml, heparinase 0.2 mg/ml, or chondroitinase ABC 2.0 units/ml. After incubation sperm cells were washed, stained with ruthenium red and studied under the electron microscope. Anionic sites in the surface of untreated spermatozoa follow regularly the plasma membrane, but present are numerous processes constituting what has been defined as the glycocalyx. Neuraminidase did not affect the distribution of ruthenium red on the surface of the spermatozoa, but eliminated almost completely the processes of the glycocalyx. Heparinase caused loss of the ruthenium red-stained sites on the membrane surface of pig spermatozoa with less influence on the dense processes of the glycocalyx. A similar loss of ruthenium red-stained sites was observed with nitrous acid treatment. A striking effect of treatment with chondroitinase ABC was the production of a typical acrosome reaction.

Animals↗