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Biomedical subjects

R Alonso

Publications and source records attributed to R Alonso.

At least 181 records · Page 10Linked to original sources

Ovarian steroids block the isoproterenol-induced elevation of pineal melatonin production in the female rat.

Changes in pineal indole metabolism during the estrous cycle, as well as in response to estrogen administration, were studied in female rats. Tyrosine hydroxylase (TH) activity, norepinephrine (NE) and indoleamine levels were determined by high-performance liquid chromatography (HPLC). Pineal melatonin was measured by radioimmunoassay (RIA). Both 5-hydroxytryptamine (5-HT) and melatonin levels, but not TH activity or NE content, were reduced during proestrus. Ovarian hormones blocked the isoproterenol-induced elevation of pineal melatonin, and reduced 5-HT levels in ovariectomized rats. These results suggest that an estrogen-induced inhibition of the pineal response to adrenergic stimulation, could be responsible for the reduction in pineal melatonin production occurring during the proestrus stage of the rat estrous cycle.

Animals↗

Characterization of two DNA populations of Giardia lamblia.

Total DNA was isolated from the parasitic protozoan Giardia lamblia and separated into two distinct populations of different densities by centrifugation through CsCl gradients containing Hoechst dye 33258. The two populations obtained were characterized by restriction enzyme analysis and nucleic acid hybridization. The less-dense population contains non-repetitive DNA and may encode mainly structural genes, such as those for alpha- and beta-tubulin. Digestion of the DNA with several restriction endonucleases showed that the denser band was composed of a 5.5 kb unit which contains the G. lamblia ribosomal RNA cistron in tandem repeated organization.

Animals↗

[3H]N-methylscopolamine binding studies reveal M2 and M3 muscarinic receptor subtypes on cerebellar granule cells in primary culture.

Saturation experiments with the muscarinic antagonist [3H]N-methylscopolamine ([3H]NMS) indicated that cerebellar granule cells in primary culture possess a high density of muscarinic acetylcholine receptors (mAChRs): Bmax = 1.85 +/- 0.01 pmol/mg of protein at 10 days in culture; KD = 0.128 +/- 0.01 nM. The selective M1 antagonist pirenzepine displaced [3H]NMS binding with a low affinity (Ki = 273 +/- 13 nM), whereas the M2/M3 muscarinic antagonist 4-diphenylacetoxy-N-methylpiperidine methiodide competed with [3H]NMS with Ki values in the nanomolar range, a result suggesting that some of the mAChRs on cerebellar granule cells belong to the M3 subtype. Methoctramine, which discriminates between M2 and M3 subtypes with high and low affinity, respectively, displayed a high and low affinity for [3H]NMS binding sites (Ki(H) = 31 +/- 5 nM; Ki(L) = 2,620 +/- 320 nM). These results provide the first demonstration that both M2 and M3 mAChR subtypes may be present on cultured cerebellar cells. In addition, complete death of neurons induced by N-methyl-D-aspartate (100 microM for 1 h) reduced by 85% the specific binding of [3H]NMS, a result indicating that most mAChRs were associated with neuronal components. Finally, the evolution of the density of mAChRs, labeled by [3H]NMS, correlated with the neuronal maturation during the in vitro development of these cells.

Animals↗

Determination of pineal melatonin by high-performance liquid chromatography with electrochemical detection: application for rhythm studies and tissue explants.

High-pressure liquid chromatography with electrochemical detection has been used for the determination of melatonin in rat pineal glands and tissue explants. Rat pineal melatonin content was measured at 2-hr intervals during a 24-hr period by direct injection of supernatants from centrifuged pineal homogenates into a reversed-phase C18 column. The lower limit of sensitivity was 32 pg per gland for a 2:1 signal-to-noise ratio, which allows melatonin measurements during daytime. Both absolute values and circadian rhythmicity obtained with the present method are in agreement with previously validated radioimmunoassays. In addition, isoproterenol-induced release of melatonin from individual rat pineal explants was measured after chloroform extraction of the incubation medium.

Animals↗

Immunoreactive levels of pineal arginine vasopressin change during the rat estrous cycle.

It has been reported that mammalian pineal activity is influenced by reproductive hormones. However, even though a relationship between serum estrogen levels and pineal indole metabolism is well documented, little is known about other pineal components. Among peptides, arginine vasopressin (AVP) has been identified in the pineal gland of several species. In the present work, variations of immunoreactive (IR)-AVP in the rat pineal during the estrous cycle were studied by radioimmunoassay. Pineal IR-AVP levels increased significantly at the proestrus afternoon, and returned to basal levels by the end of estrus. These variations were partially related to those of serum estradiol concentration. Although the functional role of AVP in the rat pineal has not been clearly elucidated, these findings provide an additional indication that it might be involved in the reproductive function of the female rat.

Analysis of Variance↗

Plasmid profiles as an epidemiological marker for Salmonella enterica serotype Enteritidis foodborne outbreaks.

The incidence of enteritidis serotype of Salmonella enterica in salmonellae infections has steadily increased in Spain from 27.1% in 1982 up to 63.4% in 1987. Given this high incidence, we have studied the plasmid profiles of Enteritidis isolates to subclassify them. Different profiles were observed in 50 isolates. In 13 Enteritidis serotype outbreaks, up to 5 different plasmid profiles were found. Each outbreak correlated with a single plasmid profile except in one case where plasmids of two different profiles were observed in strains from the same outbreak.

Bacterial Typing Techniques↗

Reproductive hormones control striatal tyrosine hydroxylase activity in the male rat.

The effects of castration, hypophysectomy and testosterone treatment on striatal tyrosine hydroxylase (TOH) activity were examined in male rats. Enzyme activity was measured by means of high-performance liquid chromatography (HPLC) determination of L-3,4-dihydroxphenylalanine (DOPA) formed. Serum levels of both testosterone and luteinizing hormone (LH) were measured by radioimmunoassay (RIA). Castration, but not hypophysectomy, reduced TOH activity in the striatum. The administration of testosterone propionate (TP) to castrated animals in a dose of 10 micrograms/100 g b.wt. during the two days previous to sacrifice, completely prevented the castration-induced reduction of striatal TOH activity. In orchidectomized rats treated with different doses of testosterone propionate (TP) up to 20 micrograms/100 g b.wt., the levels of striatal TOH activity were apparently related to either the dose-related increase of serum testosterone or the decrease of serum LH. Higher doses of the androgen failed to further modify striatal TOH activity, in spite of the dose-related elevation of serum testosterone concentration. These results suggest that circulating levels of gonadal and/or pituitary hormones partially control dopaminergic synthesis in striatal terminals, which in turn may account for some behavioral effects of reproductive hormones.

Animals↗

Partitioning behaviour in aqueous two-phase systems and fractionation by counter-current distribution of chick-embryo erythrocytes with numerical resolution of distribution curves.

The partition of chick-embryo and young-chick erythrocytes in dextran-poly-(ethylene glycol) two-phase systems depends on the interfacial tension and electrical potential differences between the phases. Counter-current distribution with charged 5% dextran-poly(ethylene glycol) systems has proved to be an adequate method for the separation of primitive and definitive erythrocytes present in chick embryos when a phase settling time of 20 min is used. The computer-aided numerical resolution of experimental curves has shown the existence of subpopulations which could not have been detected by using conventional methods.

Animals↗

New insights into maitotoxin action.

Maitotoxin (3 ng/mol) induced a massive uptake of 45Ca2+ into BC3H1 cells. This effect exhibits a lag phase of 3 min. Inositol diphosphate formation occurred concomittantly with the 45Ca2+ uptake but inositol monophosphate formation was found only after a 5-min delay following toxin addition. Maitotoxin-induced 45Ca2+ influxes could not be blocked by either 1 microM verapamil, 1 microM nifedipine or 1 mM La3+ but was blocked by Zn2+ (IC50 = 41 microM). In addition to inositol phosphate formation and 45Ca2+ uptake, maitotoxin stimulated a large uptake of Na+ and a great loss of K+ in BC3H1 cells. In the absence of Ca2+ (1 mM EGTA) none of the four maitotoxin effects could be detected. After restoration of Ca2+, the maitotoxin effects reappeared even when the toxin itself was no longer present. The divalent cation, Co2+ (1 mM), inhibited ion movements induced by maitotoxin and also digitonin (8.1 microM). The toxin action showed a very pronounced pH dependence. At low pH, maitotoxin was inactive. The dose-response curves for H+ ion inhibition of maitotoxin-induced Ca2+ uptake showed a shift to the right when determined in the absence of HCO3- and HCO3-/Cl- ions. It was concluded that the primary action of maitotoxin in BC3H1 cells was a pore-forming or channel-forming activity of a non-classical type. Some properties of maitotoxin resemble those of alpha-latrotoxin, others those of pore-forming agents such as melittin or alpha-toxin of Staphylococcus aureus.

Animals↗

Complex interactions of agonists with alpha 1-adrenoceptors in intact cells.

1. The apparent Ki values of (-)-noradrenaline (NA), (+)- and (-)-adrenaline (Ad), phenylephrine and the mono-fluorinated NAs (in position 2, 5 or 6) for alpha 1-adrenoceptors of intact BC3H1 cells labelled with [3H]-prazosin were greatly dependent on the incubation temperature. 2. The EC50 values of these compounds for stimulation of the inositol phosphate (IP) accumulation at 37 degrees C were intermediate between their apparent dissociation constants at 2 degrees C (Ki2 degrees) and at 37 degrees C (Ki37 degrees). 3. The fact that an irreversible blockade of 46% +/- 6% (n = 3) of the [3H]-prazosin binding sites by phenoxybenzamine reduced the maximal IP-formation induced by NA by 57% +/- 5% (n = 3) shows that there is a direct coupling between alpha 1-adrenoceptors and phospholipase C in BC3H1 cells. 4. The Ki37 degrees s of all agonists tested were in the same range (0.1 to 1 mM) and showed no simple correlation with their EC50 values. 5. The Ki2 degrees values for all the agonist correlated linearly with their EC50 values but were about 20-100 times lower than the respective EC50 values (except for the partial agonist methoxamine). In order to explain this difference, we propose that the apparent high affinity in the cold could be due to an [3H]-prazosin-induced alteration of the active site of the alpha 1-adrenoceptor, increasing its apparent affinity for catecholamines.

Adrenergic alpha-Agonists↗

Separation by counter-current distribution of young-chick erythrocyte populations and numerical resolution of distribution curves.

The presence of two red cell populations in young chicks has been demonstrated after increasing the settling time used during counter-current distribution with charged 5% Dextran--4% poly(ethylene glycol) two-phase systems. A Fortran program using statistical methods was applied to show the resolution in two peaks (with two or three subpopulations assumed) of the counter-current distribution curves.

Animals↗

Day-night rhythm of rat pineal tyrosine hydroxylase activity as determined by HPLC with amperometric detection.

Tyrosine hydroxylase activity in the rat pineal gland was measured by means of HPLC determination of the amount of L-3,4-dihydroxyphenylalanine formed. Enzyme activity showed a clear day-night rhythm, paralleling that of plasma melatonin levels in the same animals, with values being high during the dark period apparently because of changes in Vmax. In animals maintained under constant illumination for 3 days, tyrosine hydroxylase activity and plasma melatonin level rhythms were completely abolished, a result indicating that both are under photoperiodic control.

Animals↗

Evidence for a role of central type benzodiazepine receptors in the inhibition of the thyrotropin-releasing hormone-induced thyrotropin release from rat perifused pituitaries.

Several centrally active benzodiazepines (BZ) were tested for their ability to inhibit the TRH-induced secretion of TSH in vitro from perifused pituitaries. Diazepam, flurazepam, chlordiazepoxide (CDZ) and midazolam (10 nM) inhibited the TSH response to TRH (10 nM) by 33-50%, while medazepam, a prodrug having virtually no affinity for central BZ sites, did not. CDZ inhibition was reversed by Ro 15-1788, antagonist of the central type BZ binding sites, but not by PK 11195, antagonist of the peripheral type. The data are consistent with an involvement of central type BZ receptor sites in the TSH-lowering effects of BZ in rats.

Animals↗

Synthesis and antiviral evaluation of nucleosides of 5-methylimidazole-4-carboxamide.

Due to the antiviral activity of certain 5-substituted imidazole nucleosides related to ribavirin, 5-methylimidazole-4-carboxamide nucleosides having beta-D-ribofuranosyl, 2-deoxy-beta- and -alpha-D-ribofuranosyl, and (2-hydroxyethoxy)methyl moieties have been prepared and tested as antiviral agents. 1-beta-D-Ribofuranosyl-5-methylimidazole-4-carboxamide was obtained by deacetylation of the corresponding tri-O-acetyl nucleoside 11 or by deacetylation and ammonolysis of the blocked ethyl 5-methylimidazole-4-carboxylate nucleoside 10, which was prepared from the stannic chloride catalyzed condensation of the trimethylsilyl derivative of ethyl 4(5)-methylimidazole-5(4)-carboxylate. Glycosylation of 4(5)-methylimidazole-5(4)-carboxamide with 3,5-di-O-p-toluoyl-2-deoxy-D-erythro-pentofuranosyl chloride via mercuric cyanide method provided an anomeric mixture of the blocked 5-methylimidazole-4-carboxamide deoxynucleoside 14 along with an anomeric mixture of the 4-methyl 5-carboxamide isomer 15. Separation of compound 14 into the corresponding beta and alpha anomers was achieved by conversion to the 3',5'-di-O-acetyl derivatives 17 and 18, which after chromatographic separation were deacetylated to give 1-(2-deoxy-beta-D-erythro-pentofuranosyl)-5-methylimidazole-4-carboxa mid e and its alpha anomer 20. 1-[(2-Hydroxyethoxy)methyl]-5-methylimidazole-4-carboxamide was prepared by alkylation of the imidazole 13 with (2-acetoxyethoxy)methyl bromide followed by treatment with methanolic ammonia. All these imidazole nucleosides were tested in HeLa cell cultures against type 1 herpes simplex and vesicular stomatitis viruses. The ribofuranosyl derivative 12 showed a significant activity against type 1 herpes simplex virus.

Antiviral Agents↗

Cerebral toxoplasmosis in acquired immune deficiency syndrome.

The clinical course of and pathologic findings in a patient with cerebral toxoplasmosis and acquired immune deficiency syndrome were evaluated. The nonspecificity of CNS signs, symptoms, and laboratory features were noted. Appropriate serologic tests and a biopsy of suspicious cerebral lesions were mandatory for confirmation of the diagnosis and administration of appropriate therapy. The importance of recognizing the nature of this infectious process and of instituting vigorous therapy as promptly as possible was emphasized.

Acquired Immunodeficiency Syndrome↗