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A Quintanar

Publications and source records attributed to A Quintanar.

6 recordsLinked to original sources

Assessment of the in vitro and in vivo biological activities of the human follicle-stimulating isohormones.

Gonadotropins are synthesized and released in different molecular forms. In this article, we present evidence that the glycosylation variants of human pituitary FSH exhibit differential and divergent effects at the target cell level and that less sialylated, short-lived variants may exert significant effects in in vivo conditions. Less acidic/sialylated glycoforms (elution pH value 6.60-4.60 as disclosed by high resolution chromatofocusing of anterior glycoprotein extracts), induced higher cAMP release, estrogen production and tissue-type plasminogen activator (tPA) enzyme activity as well as cytochrome P450 aromatase and tPA mRNA expression in cultured rat granulosa cells than the more acidic analogs (pH<4.76). By contrast, the more acidic/sialylated glycoforms induced higher alpha-inhibin subunit mRNA expression than their less acidic counterparts. In cumulus enclosed oocytes isolated from mice ovaries, addition of less acidic isoforms induced resumption of meiosis more efficiently than the more acidic analogs. Interestingly, the least acidic isoform (pH>7.10) behave as a strong antagonist of several FSH-mediated effects. Assessment of the in vivo effects of the isoforms on granulosa cell proliferation in follicles from immature rats, revealed that short-lived isoforms were equally or even more efficient than their more acidic counterparts in maintaining granulosa cell proliferation when administered immediately after hypophysectomy. These results show that the naturally occurring human FSH isoforms may exhibit differential or even unique effects at the target cell level and that factors other than the metabolic clearance rate of the molecule (including receptor-binding affinity and capability of the ligand to activate its receptor and trigger intracellular signaling) also play an important role in determining the net in vivo effects of a particular FSH variant.

Animals↗

Human platelet antigen genotyping using a fluorescent SSCP technique with an automatic sequencer.

The typing of human platelet antigens (HPA) can be useful in many clinical situations such as neonatal alloimmune thrombocytopenia, post-transfusion purpura, and platelet transfusion refractoriness. The fluorescent-based single-strand conformation polymorphism (F-SSCP) technique is a fast and convenient way to perform HPA genotyping. Universal sequences from phage M13 were introduced at both ends of specific PCR-products by using 5'-tailed primers. A short second round of PCR with universal primers coupled to Cy-5 enabled the PCR-products to be fluorescently labelled. F-SSCP was performed by gel electrophoresis on an automated fluorescent DNA analyser. Genotyping of the three major HPA systems carried by the GP IIb-IIIa complex showed the F-SSCP technique to be accurate and reliable. A single gel procedure has been sufficient to detect HPA genetic polymorphisms tested to date. Neither restriction enzyme, radioactive material, nor any other hazardous chemicals such as ethidium bromide were required. This technique enabled us to genotype HPA-1, -3 and -4 alleles easily and to diagnose materno-fetal incompatibility in a rare alloantigenic system. F-SSCP is a promising technique for the detection of new mutations and/or DNA polymorphisms on a large scale.

Adult↗

A 14-kilodalton prolactin-like fragment is secreted by the hypothalamo-neurohypophyseal system of the rat.

The recently described expression of the PRL gene, and the occurrence of a 14-kilodalton (kDa)PRL-like immunoreactive protein in the hypothalamo-neurohypophyseal system of the rat have raised the possibility that PRL variants are released from neurohypophyseal terminals into the blood. In this study, we investigated the local production of a hypothalamo-neurohypophyseal 14-kDa PRL-like protein by showing an independent origin from adenohypophyseal PRL. No 14-kDa PRL-like protein was detected in adenohypophyseal extracts by Western blots, whereas chronic hypophysectomy produced no change in the immunocytochemical detection of PRLs in supraoptic and paraventricular magnocellular neurons. In addition, a 14-kDa immunoreactive PRL-like protein was released into the medium by incubated neurohypophyseal lobes. Western blot analysis showed that significantly more of this 14-kDa protein was released into calcium-containing medium (1.8 mM) than into calcium-free medium. Furthermore, depolarizing concentrations of potassium (56 mM) increased by 3-fold the release of immunoreactive PRL by incubated hypothalamo-neurohypophyseal explants. In addition, a 14-kDa PRL-like antigen was detected in the circulation of the rat by Western blot analysis. These results are consistent with the local synthesis and calcium-dependent release of neurohypophyseal PRL-like proteins that include a predominant 14-kDa form.

Animals↗

Neuroendocrine regulation of adrenal gland and hypothalamus 5'deiodinase activity. II. Effects of splanchnicotomy and hypophysectomy.

This study analyzes the role of the autonomic nervous system, the pituitary gland, ACTH, dexamethasone (DEX), and thyroid hormones in the regulation of 5'deiodinase (5'D) in the hypothalamus (HP) and adrenal gland (AG) of the rat. 5'D activity was analyzed in rats under basal conditions (22 C) and during cold exposure (4 C, during 15, 30, 60, and 120 min). Several experimental groups were formed: intact animals (INT), unilateral (left) splanchnicotomized, sham splanchnicotomized, hypophysectomized (HPX), and sham hypophysectomized. Results in the hypothalamus were: 1) independent of the experimental group, the HP 5'D activity increased during the first 15-30 min of cold exposure; however, this increase was greater in operated animals than in INT rats; and 2) basal 5'D activity was increased in HPX rats and was also regulated by thyroid hormones. Results in the adrenal gland were: 1) INT rats exhibited a biphasic pattern of 5'D activation during cold stress (30 and 60 min of exposure); 2) the splanchnic nerve exerted a tonic-stimulatory effect on basal AG 5'D activity; 3) the denervated gland preserved its ability to respond to cold; 4) in INT animals DEX but not ACTH had a stimulatory effect on basal activity; 5) the high 5'D activity post-HPX was reverted to basal values by T4 and DEX administration; 6) SHAM-HPX also was followed by a large increase in basal 5'D activity, and 7) this hyperresponse was abolished by acute ACTH and DEX administration. In conclusion, our results demonstrate that the mechanisms that participate in the regulation and activation of 5'D in the adrenal gland and the hypothalamus are of a neuroendocrine nature. Also, in both organs, but mainly in the HP, 5'D activity is T4-dependent. In addition to the tonic-stimulatory influence conveyed by the splanchnic nerve, AG 5'D activity is influenced by thyroid hormones, glucocorticoids, and probably extrapituitary factors whose nature is unknown yet.

Adrenal Glands↗

On the nature of the follicle-stimulating signal delivered to the ovary during exogenously controlled follicular maturation. A search into the immunological and biological attributes and the molecular composition of two preparations of urofollitropin.

In the present study, we analyzed the immunological and biological potencies as well as the molecular composition of urinary follicle-stimulating hormone (FSH) present in determined lots of regular and highly purified (HP) commercial preparations of urofollitropin in order to obtain additional insights on the particular type of gonadotropin signal received by the ovary during exogenously regulated ovarian stimulation. In both preparations, a high degree of FSH charge heterogeneity was detected as disclosed by chromatofocusing analysis (pH range 7.5 to < 4.0). Urinary FSH present in the HP compound was consistently more acidic and exhibited a longer survival in rat circulation than the regular formulation. Inter-batch variability for FSH heterogeneity and in vitro bioactivity was higher in the partially purified preparation than in the HP analog. In the regular preparation, the amount of immunoreactive and bioactive FSH per ampule was two times higher than that present in the HP preparation; the resultant in vitro B/I ratios were similar. Although both urinary FSH preparations showed detectable amounts of immunoreactive and bioactive luteinizing hormone and choriogonadotropin hormone material, the degree of activity present in the less purified formulation was considerably higher than that shown by the HP analog. When the capability of each urinary FSH preparation to induce ovarian tissue-type plasminogen activator enzyme activity in hypophysectomized rats was determined, both formulations exhibited similar potencies despite the existing differences in plasma clearance rate and charge distribution profile. The present study indicates that the isoform composition of urinary FSH in the two commercial preparations analyzed differs according to the degree of purity of the formulation. More FSH material is needed in the partially purified FSH preparation to induce biological effects similar in magnitude to those exhibited by the highly purified analog. The possible impact of these variations in the molecular composition of the FSH signal on other biological functions of the ovary during the course of exogenously controlled follicular growth and maturation still remains to be ascertained.

Animals↗

[Wood anatomy and uses of 7 tropical tree species from Mexico].

The wood anatomy of Coccoloba cozumelensis Hemsl., Coccoloba spicata Lundell, Gymnanthes lucida Sw., Blomia cupanioides Miranda, Canella winterana (L.) Gaertn., Aspidosperma megalocarpon Müell Arg. and Ehretia tinifolia L., is described. One tree per species was collected in the tropical rain forest of Quintana Roo, Mexico. Their wood has important traditional uses in furniture, tools, rural buildings, posts, fences, railroads and firewood. Macroscopic and microscopic characteristics were described and measured in wood samples, permanent slides and macerated material. These species have diffuse porosity, alternate vessel pits, simple perforation plates, numerous and small rays; libriform fibres are common, as well as ergastic material in gum forms, calcium carbonate and silica crystals. These characteristics explain aesthetical, weight, hardness and resistance (to mechanical and biological damage) characters that fit traditional use by the Maya.

Classification↗