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Biomedical subjects

E Rojas

Publications and source records attributed to E Rojas.

At least 91 records · Page 5Linked to original sources
PubMed1995

Oxotremorine-m potentiation of glucose-induced insulin release from rat islets involves M3 muscarinic receptors.

cDNAs encoding for M1 and M3 muscarinic acetylcholine (ACh) receptors were detected in rat pancreatic islet cells by polymerase chain reaction (PCR) amplification techniques. A new cholinergic agonist, oxotremorine-m (oxo-m), in the presence of glucose (5.6 mM), produced a dose-dependent potentiation of insulin secretion saturating at approximately 5 microM. This effect was suppressed by the L-type Ca2+ channel blocker nifedipine. Higher doses of oxo-m (50 microM) induced a biphasic insulin response both at low (5.6 mM) or high (16.7 mM) glucose concentrations. In a Ca(2+)-deficient medium containing glucose (5.6 mM), oxo-m evoked only a reduced first phase of insulin secretion. The potentiating effects of oxo-m were inhibited by the muscarinic receptor antagonists 4-diphenylacetoxy-N-methylpiperidine methiodide (M3), hexahydro-sila-difenidol hydrochloride, p-fluoro analogue (M3 > M1 > M2), and pirenzepine (M1) in a dose-dependent manner; half-maximal inhibitory concentration values were approximately 5, 20, and 340 nM, respectively. The PCR results demonstrate the presence of M1 and M3 muscarinic ACh receptors in the islet tissue, and the secretion data strongly suggest that the potentiation of glucose-induced insulin release evoked by oxo-m depends on the activation of a muscarinic M3-subtype receptor present in the beta-cell membrane.

Animals↗
PubMed1995

[Leishmania braziliensis: isolation from lesions by inoculation of hamsters with and without addition of salivary gland lysate from Lutzomyia youngi].

Homogenized biopsy tissue from the cutaneous leishmaniasis lesions of 50 patients from Trujillo, Venezuela, were inoculated subcutaneously into the tarsi of male hamsters. Homogenized tissue either alone or mixed with salivary gland lysates of Lutzomyia youngi were used for inoculation. Homogenized tissue alone yielded 58.5% of infections with a mean of twelve weeks for prepatency, while those mixed with sandfly lysate resulted in 92% of infections with a mean prepatency of three weeks.

Adolescent↗
PubMed1995

[Excretion of promastigotes of Leishmania pifanoi by experimentally infected Lutzomyia youngi].

The increase in the promastigotes population of Leishmania pifanoi in Lutzomyia youngi experimentally infected and kept on 50% sacarose under constant conditions of temperature and humidity is described. Two stages in the differentiation and growth of the parasites are recognised between two and twenty-four hours after meals. The pleomorphic differentiation of the amastigotes in short promastigotes which multiply by binary division for 60 hours, when the rupture of the peritrophic membrane occurs, takes place within 48 hours. The second stage occurs between 72 and 96 hours when some of the parasites migrate to the esophagic valve and the rest of the free parasites are excreted in fecal drops as large, active promastigotes. The first drops excreted react positively to glucose or contain crystals of urate. The excess promastigotes of the second stage of development are eliminated in the last excretions and react positively to the Hemoscreen and Biuret tests for total proteins as also for glucose and account for 82% of the of drops excreted. The excretion of parasites by Lu. youngi is a normal stage in the growth of L. pifanoi into a vector.

Animals↗
PubMed1995

Modulatory mechanism of ACTH on insulin secretion: effect on cytosolic Ca2+, membrane potential and Ca(2+-ATPase activity.

The aim of this work was to get some insight into the mechanism by which ACTH produces its enhancing effect on glucose-induced insulin secretion. For this purpose we have determined: a) the release of insulin by isolated rat islets incubated with 3.3 or 16.6 mM glucose with or without the addition of 500 pg/ml ACTH, together with the changes induced by ACTH on b) cytosolic [Ca2+] of isolated B cells, c) islet plasma membrane Ca(2+)-ATPase activity and d) changes in membrane potential of single mouse islets. ACTH significantly enhanced the release of insulin elicited by either 3.3 or 16.6 mM glucose. This hormone concentration also induced a significant increase in the cytosolic [Ca2+] in isolated B cells. ACTH did not produce B cell membrane depolarization. Conversely, ACTH produced a significant decrease in islet plasma membrane Ca(2+)-ATPase activity. These results suggest that ACTH in concentrations similar to those attained by the endogenous peptide at the islet interstitium exerts its positive modulation on glucose-induced secretion of insulin, at least partly through its increasing effect on cytosolic [Ca2+] of B cells. The latter might be the consequence of the decreasing effect of ACTH on Ca(2+)-ATPase activity rather than to stimulation of voltage-dependent Ca(2+)-channels.

Adrenocorticotropic Hormone↗
PubMed1995

Effects of selected synthetic compounds on growth of Mucor rouxii.

The in vitro activity of four synthetic compounds was tested on fungal cells from Mucor rouxii. The compounds included phenylenediamine, two phenolamines, and quinone. At the concentrations tested (10(-2)-10(-4)M) the compounds exhibited antifungal activity, with the exception of quinone. On the basis of their effects on spore germination, and development of hyphae, phenylenediamine was the more active. The mechanism of action of the compounds is still unknown, but hyphae show morphological alterations and disturb the distribution of calcofluor in the cell wall. This suggests variations in the genesis of the cell wall.

Aniline Compounds↗
PubMed1995

Identification of mammalian noggin and its expression in the adult nervous system.

The multiple roles of noggin during dorsal fate specification in Xenopus embryos, together with noggin's ability to directly induce neural tissue, inspired an effort to determine whether a similar molecule exists in mammals. Here we describe the identification of human and rat noggin and explore their expression patterns; we also localize the human NOGGIN gene to chromosome 17q22, and the mouse gene to a syntenic region of chromosome 11. Mammalian noggin is remarkably similar in its sequence to Xenopus noggin, and is similarly active in induction assays performed on Xenopus embryo tissues. In the adult mammal, noggin is most notably expressed in particular regions of the nervous system, such as the tufted cells of the olfactory bulb, the piriform cortex of the brain, and the Purkinje cells of the cerebellum, suggesting that one of the earliest acting neural inducers also has important roles in the adult nervous system.

Amino Acid Sequence↗
PubMed1994

beta-Amyloid Ca(2+)-channel hypothesis for neuronal death in Alzheimer disease.

The Alzheimer's Disease (AD) amyloid protein (A beta P[1-40]) forms cation selective channels when incorporated into planar lipid bilayers by fusion with liposomes containing the peptide. Since the peptide has been proposed to occur in vivo in both membrane-bound and soluble forms, we also tested the possibility of direct incorporation of the soluble A beta P[1-40] into the membrane. We found the peptide can also form similar channels in acidic phospholipid bilayers formed at the tip of a patch pipet, as well as in the planar lipid bilayer system. As in the case of liposome mediated incorporation, the A beta P[1-40]-channel in the solvent-free membrane patch exhibits multiple cation selectivity (Cs+ > Li+ > Ca2+ > or = K+), and sensitivity to tromethamine. The fact that equivalent A beta P[1-40] amyloid channels can be detected by two different methods thus provides additional validation of our original observation. Further studies with a beta P-channels incorporated into planar lipid bilayers from the liposome complex have also revealed that the channel activity can express spontaneous transitions to a much higher range of conductances between 400 and 4000 pS. Under these conditions, the amyloid channel continues to be cation selective but loses its tromethamine sensitivity. By contrast, amyloid channels were insensitive to nitrendipine at either conductance range. We calculate that if such channels were expressed in cells, the ensuing ion fluxes down their electrochemical potential gradients would disrupt cellular homeostasis. We therefore interpret these data as providing further support for our beta-amyloid Ca(2+)-channel hypothesis for neuronal death in Alzheimer's Disease.

Alzheimer Disease↗
PubMed1994

The diabetogenic agent alloxan increases K+ permeability by a mechanism involving activation of ATP-sensitive K(+)-channels in mouse pancreatic beta-cells.

The effects of the diabetogenic agent, alloxan, on membrane potential, input resistance and electrical activity of normal mouse pancreatic beta-cells were studied. Tetraethylammonium (TEA), quinine and Glyburide were used to block K(+)-channels and to elucidate the mechanisms underlying alloxan's effects on beta-cell membrane potential. Exposure of the islet to alloxan (75-100 microM) in the presence of glucose (11 mM), produced a rapid (15 sec), transient inhibition of electrical activity, often accompanied by hyperpolarization of the membrane, and this was followed by recovery of the burst pattern. This early effect of alloxan was followed after approximately 15 min by a complete inhibition of electrical activity and hyperpolarization. The inhibition accompanied by hyperpolarization was associated with a decrease in input resistance, indicating increased K(+)-conductance. Both the transient and delayed effects of alloxan were blocked by glucose (33 mM), quinine and glyburide but not by other conditions which induce continuous electrical activity such as elevated external [K+] (10 mM), ouabain, K+ removal, or TEA (20 mM). The transient inhibition induced by alloxan may be due to a direct competition with glucose transport/metabolism since it did not occur when alpha-keto isocaproic acid (KIC) was used to induce electrical activity. The delayed inhibition may reflect indirect effects of accumulation of this agent or its metabolites within the cell. Since both effects of alloxan are blocked by glyburide they appear to involve activation of the ATP-sensitive K(+)-channel (K-ATP).

Alloxan↗
PubMed1994

Evaluation of the carcinogenic and genotoxic potential of praziquantel in the Syrian hamster embryo cell transformation assay.

Praziquantel, a drug used for the treatment of neurocysticercosis, was tested for its ability to induce morphological transformation of Syrian hamster embryo fibroblasts. Results indicate that praziquantel transforms these cells without affecting their viability. Further experiments were carried out to investigate its possible mechanism of action in the same cell system. Micronucleus formation was observed in cultures treated with concentrations which induced morphological transformation, about 40% of these micronuclei were positive to a kinetochore antibody. No induction of DNA repair synthesis was observed even at cytotoxic concentrations. These results suggest that praziquantel has an aneugenic effect which could be responsible for its ability to transform morphologically these cells. Risk-benefit analysis should be carried out whenever this drug is utilized.

Animals↗
PubMed1994

Membrane potential regulates inositol 1,4,5-trisphosphate-controlled cytoplasmic Ca2+ oscillations in pituitary gonadotrophs.

The influence of membrane potential (Vm) on cytoplasmic calcium ([Ca2+]i) oscillations during the sustained extracellular Ca(2+)-dependent phase of the Ca2+ signaling response to gonadotropin-releasing hormone (GnRH) was analyzed in cultured pituitary gonadotrophs. In agonist- and inositol (1,4,5)-trisphosphate (Ins(1,4,5)P3)-stimulated cells, sustained [Ca2+]i oscillations were extinguished by hyperpolarization after 3-15 min despite the availability of Ca2+ in the extracellular medium. Single depolarizing pulses transiently restored the amplitude of the sustained spiking in a dihydropyridine- and extracellular Ca(2+)-sensitive manner. The responses to depolarization showed a marked dependence on Vm that was correlated with the steady-state inward Ca2+ current. In addition, repetitive application of brief depolarizing pulses modulated the frequency of agonist- and Ins(1,4,5)P3-controlled spiking; depolarization pulses at frequencies lower than the intrinsic rate of episodic Ca2+ release triggered large transients between the autonomous spikes, whereas higher frequencies of depolarizing pulses overcame the original Ca2+ spiking frequency. These extrinsically driven and extracellular Ca(2+)-dependent oscillations were sensitive to the Ca(2+)-ATPase blocker, thapsigargin, but not to ryanodine. On the other hand, spontaneous firing and application of depolarizing pulses to nonstimulated cells failed to induce thapsigargin-sensitive oscillations. These findings demonstrate that the pattern of Ca2+ signaling in gonadotrophs does not depend exclusively on the Ins(1,4,5)P3 concentration, but also on the excitable status of the cell. Such modulation of the Ins(1,4,5)P3-controlled Ca2+ signaling system by changes in Vm could provide a mechanism for the integration of multiple inputs that utilize diverse signal transduction pathways.

Action Potentials↗
PubMed1994

Intracellular Ca2+ transients induced by high external K+ and tetracaine in cultured rat myotubes.

Cultured myotubes from rat neonatal skeletal muscle were used to measure intracellular Ca2+ concentration ([Ca2+]i) and membrane potentials (Vm) using the Indo-1 microfluorimetry method and the nystatin perforated membrane patch technique, respectively. Sudden increases in external [K+]o from 5 mM to either 22, 42 or 84 mM elicited transient elevations in [Ca2+]i from a resting level of 106.2 +/- 10.3 nM (n = 41) to peak values of 297, 409 and 454 nM, respectively. Vm changes induced by elevated [K+]o followed the Nernst equation for [K+]o. The complex Ca2+ release response induced by elevated [K+]o can be described by a minimal model involving two components with different kinetics. This analysis revealed that the extent of the Ca2+ release by the fast component bears a sigmoidal relationship with Vm (midpoint at -47.5 mV and an effective valence of 4). Furthermore, while the fast transitory component was rather insensitive to [Ca2+]o and nifedipine, the slow component was profoundly inhibited by the dihydropyridine (10 microM) both in normal and in a Ca2+ deficient medium. Tetracaine (0.05 to 2 mM), a blocker of the charge movement associated with excitation-contraction (E-C) coupling, elicited a fast elevation in [Ca2+]i followed by a rise at a constant rate to levels as high as 1-2 microM, and the changes in [Ca2+]i were readily reversible. Simultaneous measurements of Vm and [Ca2+]i suggest that the fast component is coupled to the rapid depolarization of the membrane induced by the anesthetic. We concluded that tetracaine triggers the release of Ca2+ from internal stores by at least two different mechanisms, one of which is associated with the depolarizing effects of the drug.

Animals↗
PubMed1994

Rise in cytoplasmic Ca2+ induced by monensin in bovine medullary chromaffin cells.

Monensin, a Na+/H+ exchanger, induces catecholamine secretion from adrenal chromaffin cells by an unknown mechanism. We found and report here that in bovine chromaffin cells, monensin evokes profound changes in [Ca2+]i which were measured by means of the fluorescent Ca2+ indicator Indo-1. Application of monensin (10 microM) generated a marked [Ca2+]i rise. Removal of external Ca2+ did not prevent the elevation of [Ca2+]i, though it was significantly decreased. In the presence of nifedipine (10 microM) or tetrodotoxin (3 microM) the monensin-induced [Ca2+]i rise remained unchanged. In contrast, in the absence of extracellular Na+ the [Ca2+]i rise was abolished. Addition of caffeine (40 mM) at the peak response generated by monensin produced a further increase in [Ca2+]i, which was independent of external [Ca2+] or [Na+]. After depletion of the IP3-sensitive compartment by thapsigargin (1 microM), caffeine still induced a rise in [Ca2+]i while the monensin response was absent. We concluded that the origin of the Ca2+ for the [Ca2+]i increase elicited by the Na+/H+ exchanger in chromaffin cells is not the extracellular space. Clearly there seems to be at least two intracellular Ca2+ stores, one of which is affected by monensin. This Ca2+ pool, which is different than the pool stimulated by caffeine, is sensitive to the extracellular [Ca2+] and to thapsigargin. Our data are compatible with the idea that the monensin mediated Na+ entry could activate the production of inositol trisphosphate and this in turn could trigger Ca2+ release from the endoplasmic reticulum.

Adrenal Medulla↗
PubMed1994

A geometric sequence that accurately describes allowed multiple conductance levels of ion channels: the "three-halves (3/2) rule".

Ion channels can express multiple conductance levels that are not integer multiples of some unitary conductance, and that interconvert among one another. We report here that for 26 different types of multiple conductance channels, all allowed conductance levels can be calculated accurately using the geometric sequence gn = g(o) (3/2)n, where gn is a conductance level and n is an integer > or = 0. We refer to this relationship as the "3/2 Rule," because the value of any term in the sequence of conductances (gn) can be calculated as 3/2 times the value of the preceding term (gn-1). The experimentally determined average value for "3/2" is 1.491 +/- 0.095 (sample size = 37, average +/- SD). We also verify the choice of a 3/2 ratio on the basis of error analysis over the range of ratio values between 1.1 and 2.0. In an independent analysis using Marquardt's algorithm, we further verified the 3/2 ratio and the assignment of specific conductances to specific terms in the geometric sequence. Thus, irrespective of the open time probability, the allowed conductance levels of these channels can be described accurately to within approximately 6%. We anticipate that the "3/2 Rule" will simplify description of multiple conductance channels in a wide variety of biological systems and provide an organizing principle for channel heterogeneity and differential effects of channel blockers.

Animals↗
PubMed1994

Theoretical models of the ion channel structure of amyloid beta-protein.

Theoretical methods are used to develop models for the ion channel structure of the membrane-bound amyloid beta-protein. This follows recent observations that the beta-protein forms cation-selective channels in lipid bilayers in vitro. Amyloid beta-protein is the main component of the extracellular plaques in the brain that are characteristic of Alzheimer's disease. Based on the amino acid sequence and the unique environment of the membrane, the secondary structure of the 40-residue beta-protein is predicted to form a beta-hairpin followed by a helix-turn-helix motif. The channel structures were-designed as aggregates of peptide subunits in identical conformations. Three types of models were developed that are distinguished by whether the pore is formed by the beta-hairpins, the middle helices, or by the more hydrophobic C-terminal helices. The latter two types can be converted back and forth by a simple conformational change, which would explain the variable conduction states observed for a single channel. It is also demonstrated how lipid headgroups could be incorporated into the pore lining, and thus affect the ion selectivity. The atomic-scale detail of the models make them useful for designing experiments to determine the real structure of the channel, and thus further the understanding of peptide channels in general. In addition, if beta-protein-induced channel activity is found to be the cause of cell death in Alzheimer's disease, then the models may be helpful in designing counteracting drugs.

Alzheimer Disease↗
PubMed1994

Presence of Leishmania braziliensis in blood samples from cured patients or at different stages of immunotherapy.

By using a PCR assay specific for Leishmania braziliensis in blood samples, we detected this parasite in patients cured by immunotherapy or at different stages of treatment. We also found the parasite in subjects who had never suffered leishmaniasis but who had lived in endemic areas and migrated to nonendemic ones many years ago. These results suggest that L. braziliensis infections are difficult to eradicate and that a clinical cure but rarely a complete elimination of the parasite is generally accomplished.

Animals↗
PubMed1994

Control of cytosolic free calcium in cultured human pancreatic beta-cells occurs by external calcium-dependent and independent mechanisms.

Changes in cytosolic intracellular free Ca2+ ([Ca2+]i) in response to glucose, glyburide, cholinergic agonists, and elevated [K+]o (external potassium concentration) were measured in cultured human islet beta-cells. In the absence of glucose, the mean resting [Ca2+]i in single beta-cells was 84.5 +/- 4.7 nM (n = 86) and remained unchanged in low external [Ca2+]o (Ca2+ concentration) (< 0.2 microM) at 23-25 C. Glucose (5.6-33 mM) induced a slow dose-related [Ca2+]i rise up to 300.0 +/- 50.6 nM (n = 19). This [Ca2+]i rise always occurred with a delay that varied from cell to cell (approximately 10-120 sec), and the steady state [Ca2+]i exhibited a sigmoidal dependence on glucose concentration (midpoint at 14.9 mM). The glucose-induced rise in [Ca2+]i was attenuated by about 62% in low external [Ca2+]o and was not affected by dantrolene, a drug that inhibits Ca2+ release from the endoplasmic reticulum. In the absence or presence of glucose, cholinergic receptor agonists evoked a biphasic increase in [Ca2+]i up to 350 nM; the delayed component of the [Ca2+]i rise was blocked by dantrolene. A rapid elevation of [K+]o to 40 mM also elicited a biphasic rise in [Ca2+]i, which peaked at about 250 nM and was inhibited by the Ca2+ channel antagonist nifedipine. Glyburide (4 microM) in the absence of glucose also induced a [Ca2+]o-dependent rise in [Ca2+]i. Increasing the concentration of glucose from 4 to 16.7 mM evoked a biphasic pattern of insulin secretion from perifused isolated islets at 37 C. Finally, in the presence of 4 mM glucose, a cholinergic muscarinic receptor agonist stimulated insulin secretion. A glucose-stimulated [Ca2+]i rise was also studied at 24 and 37 C in cultured rat islet cells. Our results suggest that the Ca2+ required for glucose-induced and muscarinic agonist-potentiated insulin release enters the cytosol from both extracellular and intracellular Ca2+ stores.

Calcium↗
PubMed1994

[Chagas' disease among blood donors in Bolivia].

A questionnaire survey of blood donors at Santa Cruz General Hospital, Bolivia, showed that while there were no seropositive cases of syphilis, HBsAg, and HIV, the prevalence of Chagas' disease was very high (23%) among the 225 blood donors who responded to this questionnaire. Actual cases of Chagasic seropositive blood being used for blood transfusion were seen, including the urgent need for a program for Chagas' disease in Bolivia. From the results of this study, it is recommended that for blood donors from South American countries, the presence/absence of Chagas' disease should be confirmed.

Adolescent↗