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C M Carvalho

Publications and source records attributed to C M Carvalho.

45 records · Page 3Linked to original sources
PubMed1993

Relation of exocytotic release of gamma-aminobutyric acid to Ca2+ entry through Ca2+ channels or by reversal of the Na+/Ca2+ exchanger in synaptosomes.

The specific inhibitor of the gamma-aminobutyric acid (GABA) carrier, NNC-711, (1-[(2-diphenylmethylene)amino]oxyethyl)- 1,2,5,6-tetrahydro-3-pyridine-carboxylic acid hydrochloride, blocks the Ca(2+)-independent release of [3H]GABA from rat brain synaptosomes induced by 50 mM K+ depolarization. Thus, in the presence of this inhibitor, it was possible to study the Ca(2+)-dependent release of [3H]GABA in the total absence of carrier-mediated release. Reversal of the Na+/Ca2+ exchanger was used to increase the intracellular free Ca2+ concentration ([Ca2+]i) to test whether an increase in [Ca2+]i alone is sufficient to induce exocytosis in the absence of depolarization. We found that the [Ca2+]i may rise to values above 400 nM, as a result of Na+/Ca2+ exchange, without inducing release of [3H]GABA, but subsequent K+ depolarization immediately induced [3H]GABA release. Thus, a rise of only a few nanomolar Ca2+ in the cytoplasm induced by 50 mM K+ depolarization, after loading the synaptosomes with Ca2+ by Na+/Ca2+ exchange, induced exocytotic [3H]GABA release, whereas the rise in cytoplasmic [Ca2+] caused by reversal of the Na+/Ca2+ exchanger was insufficient to induce exocytosis, although the value for [Ca2+]i attained was higher than that required for exocytosis induced by K+ depolarization. The voltage-dependent Ca2+ entry due to K+ depolarization, after maximal Ca2+ loading of the synaptosomes by Na+/Ca2+ exchange, and the consequent [3H]GABA release could be blocked by 50 microM verapamil. Although preloading the synaptosomes with Ca2+ by Na+/Ca2+ exchange did not cause [3H]GABA release under any conditions studied, the rise in cytoplasmic [Ca2+] due to Na+/Ca2+ exchange increased the sensitivity to external Ca2+ of the exocytotic release of [3H]GABA induced by subsequent K+ depolarization. Thus, our results show that the vesicular release of [3H]GABA is rather insensitive to bulk cytoplasmic [Ca2+] and are compatible with the view that GABA exocytosis is triggered very effectively by Ca2+ entry through Ca2+ channels near the active zones.

Animals↗
PubMed1991

Regulation of carrier-mediated and exocytotic release of [3H]GABA in rat brain synaptosomes.

In this study we investigated the role of external monovalent cations, and of intracellular Ca2+ concentration ([Ca2+]i) in polarized and depolarized rat cerebral cortex synaptosomes on the release of [3H]-gamma-aminobutyric acid (3H-GABA). We found that potassium-depolarization, in the absence of Ca2+, of synaptosomes loaded with 3H-GABA releases 7.4 +/- 2.1% of the accumulated neurotransmitter, provided that the external medium contains Na+, and an additional 19.0 +/- 2.5% is released upon adding 1.0 mM CaCl2 to the exterior. The Ca(2+)-independent release component does not occur in a choline medium and it is only 3.4 +/- 0.8% of the 3H-GABA accumulated in a Li+ medium, but both ions support the Ca(2+)-dependent release of 3H-GABA (13.4 +/- 0.6% in choline and 15.4 +/- 1.5% in Li+), which suggests that the exocytotic release is independent of the external monovalent cation present, whereas the carrier-mediated release specifically requires Na+ outside. Furthermore, previous release of the cytosolic 3H-GABA due to predepolarization in the absence of Ca2+ does not influence the amount of 3H-GABA subsequently released by exocytosis due to Ca2+ addition (19.1 +/- 2.5% or 19.1 +/- 1.1%, respectively). In choline or Li+ medium, the value of the [Ca2+]i is raised by Na+/Ca2+ exchange to 663 +/- 75 nM or 782 +/- 54 nM, respectively, within three minutes after adding 1.0 mM Ca2+, in the absence of depolarization, and parallel release experiments show no release of 3H-GABA in the choline medium, but a substantial release (7.1 +/- 2.1%) of 3H-GABA occurs in the Li+ medium without depolarization. Subsequent K(+)-depolarization shows normal Ca(2+)-dependent release of 3H-GABA in the choline medium (14.1 +/- 2.0%) but only 8.6 +/- 1.1% release in the Li+ medium, which suggests that raising the [Ca2+]i by Na+/Ca2+ exchange, without depolarization, supports some exocytotic release in Li+, but not in choline media. The role of [Ca2+]i and of membrane depolarization in the release process is discussed on the basis of the results obtained and other relevant observations which suggest that both Ca2+ and depolarization are essential for optimal exocytotic release of GABA.

Animals↗
PubMed1991

Calcium stores in electropermeabilized HL-60 cells before and after differentiation.

Non-induced HL-60 cells (N-IND) and HL-60 cells induced to differentiate with 2 microM retinoic acid (IND) were electropermeabilized with electrical discharges, and the intracellular Ca2+ stores were measured in each type of cell. Both N-IND and IND cells accumulate Ca2+ in the presence of ATP after electropermeabilization. The Ca2+ is stored in at least two different compartments; accumulation in one of the compartments is inhibited by oligomycin and CCCP, and it is not releasable by Ins(1,4,5)P3. The maximal accumulation of Ca2+ by the Ins(1,4,5)P3 sensitive pool is about 0.3 nmol/10(6) cells and 0.9 nmol/10(6) cells for the N-IND and for the IND cells, respectively, and the half-maximal value occurs at a free Ca2+ concentration of 0.23 microM and 0.63 microM, respectively. The oligomycin + CCCP sensitive pool hardly accumulates any Ca2+ at this level of free Ca2+, but at higher free [Ca2+] (greater than microM) its maximal capacity is 80-100-fold higher than the Ins(1,4,5)P3-sensitive pool (about 17-18 nmol/10(6) cells). It is concluded that at physiological free Ca2+ concentrations, the non-mitochondrial Ca2+ pool is regulating the intracellular free Ca2+ in N-IND and IND HL-60 cells, and that this Ca2+ pool can be mobilized by Ins(1,4,5)P3. Furthermore, the capacity of this pool increases about 3-fold when the cells are induced to differentiate with retinoic acid.

Adenosine Triphosphate↗
PubMed1989

Partition of Ca2+ antagonists in brain plasma membranes.

The partition coefficients (Kp) of three prototype Ca2+ antagonists, nitrendipine, (-)-desmethoxyverapamil and flunarizine were determined in native synaptic plasma membranes (SPM) isolated from sheep brain cortex and in liposomes prepared with the total lipids extracted from the membranes. We found that at 25 degrees and at 5 x 10(-6) M drug concentration the Kp values of the drugs for native SPM are higher than those obtained for liposomes, and are of the order of 334 +/- 53, 257 +/- 36 and 23 X 10(3) for nitrendipine, (-)desmethoxyverapamil and flunarizine, respectively, whereas the Kp values in liposomes are 190 +/- 41, 118 +/- 10 and 6 x 10(3) for the same drugs. The results suggest that the presence of membrane proteins favors the incorporation of the drugs in the membranes. Furthermore, the Kp values of the three Ca2+ antagonists studied increase with temperature in native membranes, but not in liposomes. It is concluded that the physical partitioning in membranes of drugs which act on Ca2+ channels may play some role in the mechanism of interaction of these drugs with the Ca2+ channel proteins.

Animals↗
PubMed1986

gamma-Aminobutyric acid release from synaptosomes as influenced by Ca2+ and Ca2+ channel blockers.

We studied the correlation between the high affinity binding of Ca2+ channel blockers to purified synaptic plasma membranes (SPM) and the effect of these drugs in blocking the 45Ca2+ uptake and the release of [3H]gamma-aminobutyric acid [( 3H]GABA) by preloaded synaptosomes. The Ca2+ channel blocker binding sites were characterized by studying the binding of the dihydropyridine, [3H]nimodipine, and of the phenylalkylamine, (-)-[3H]desmethoxyverapamil, to purified SPM isolated from sheep brain cortex synaptosomes. The purified SPM had high affinity binding sites for both Ca2+ channel blockers. The binding parameters were similar to those previously reported for whole brain homogenates: KD = 0.64 nM and Bmax = 160 fmol/mg of protein for [3H]nimodipine, and KD = 7.9 nM and Bmax = 1,500 fmol/mg of protein for (-)-[3H]desmethoxyverapamil. The Ca2+ channel blockers inhibited the release of [3H]GABA induced by K+ depolarization in the presence or in the absence of Ca2+. The Ca2+-dependent component of [3H]GABA release was inhibited by verapamil, (-)-D 600, d-cis-diltiazem, nifedipine and PY 108-86 with IC50 values of 2.2 X 10(-5) M, 6.3 X 10(-5) M, 3 X 10(-4) M, greater than 10(-4) M and 3 X 10(-5) M, respectively. Furthermore, the Ca2+ channel blockers also inhibited the Ca2+-independent [3H]GABA release which occurred in the presence, but not in the absence, of external Na+. The Ca2+ channel blockers at concentrations which inhibited [3H]GABA release inhibited the entry of Ca2+ through the Ca2+ channels and also the entry of Ca2+ by Na+/Ca2+ exchange. We conclude that the concentrations of Ca2+ blockers necessary to block Ca2+ uptake through the Ca2+ channels and by Na+/Ca2+ exchange coincide with the concentrations at which they inhibit [3H]GABA release, but that their effect on the relationship between Ca2+ uptake and [3H]GABA release is different for the various blockers. The effects of the drugs on Ca2+ movements and [3H]GABA release are not specifically mediated through the high affinity binding of the drugs since relatively high concentrations were necessary (greater than 10(-5) M) for the effects reported here.

Animals↗
PubMed1984

Occurrence and experimental infection of toads (Bufo marinus and B. granulosus) with Mycobacterium chelonei subsp. abscessus.

In a survey of 234 Amazonian toads and frogs, six strains of Mycobacterium chelonei subsp. abscessus were isolated from the liver or spleen of four of 66 Bufo marinus (6.1%) and from the kidney or peritoneal fluid of two of 86 B. granulosus (2.3%). There were no histopathological lesions in the viscera of the infected animals. Experimental infection of 29 captive B. marinus and B. granulosus, by the intraperitoneal route, with a pooled inoculum of M. chelonei subsp. abscessus caused five deaths near the end of a 2-month observation period. M. chelonei subsp. abscessus was isolated from the liver, spleen, kidney, gonad, heart and lung of toads killed at various intervals after inoculation, and intracellular acid-fast bacilli were seen in these organs. Histological evidence of invasion of tissues by mycobacteria became apparent from the 45th day after infection. The susceptibility to infection of B. marinus and B. granulosus suggests that these toads may serve as a fortuitous animal host for M. chelonei subsp. abscessus.

Animals↗
PubMedDate not supplied

Regulation of intracellular [Ca2+] and GABA release by presynaptic GABAB receptors in rat cerebrocortical synaptosomes.

In this study we determined the changes in the intracellular free Ca2+ concentration, associated with the inhibitory modulation of the exocytotic release of GABA by GABAB receptor activation in rat cerebrocortical synaptosomes. We observed that SK&F 97541 and (-)baclofen both act as agonists of the presynaptic GABAB receptors in modulating GABA release and Ca2+ influx due to KCl (10 mM) depolarization, but SK&F 97541 is more potent than (-)baclofen in modulating both Ca2+ influx and GABA release. Thus, activation of GABAB receptors by either SK&F97541 (10 microM) or by (-)baclofen (100 microM) caused about 18% inhibition of the increase in [Ca2+]i, due to KCl depolarization, and inhibited the [3H]GABA release by about 30%. The pharmacological similarities of the GABAB receptor activation in producing inhibition of both calcium channel mediated influx of Ca2+ and transmitter release suggest that presynaptic inhibition of GABA release by GABAB receptor activation may result, at least in part, from inhibition of Ca2+ influx through P-type (or possibly Q-type) Ca2+ channels, sensitive to omega-Agatoxin IVA (200 nM). Furthermore, modulation of GABA release of GABAB receptors was abolished by preincubation with pertussis toxin, suggesting that a pertussis toxin sensitive G protein may be the coupling factor between GABAB receptors and the voltage-dependent Ca2+ channels associated with the exocytotic release of GABA in rat cerebrocortical nerve terminals.

Animals↗
PubMedDate not supplied

[Nursing care: its meaning to nurses and patients].

The objective of this research is to clarify the concept of nursing care through the analysis of the discourse of nurses and patients. The data were collected in a clinic of a university hospital in Fortaleza-Ceará, in January 2001. The sampling frame was categorized through forms, and the speech of nurses and patients was obtained through semi-structured interviews. Data were formatted into textual form and analyzed according to the discourse analysis method. In the following phase, results were interpreted based on hermeneutic principles. Results showed that nursing professionals link the concept of care to the satisfaction of patients' needs and to their physical and emotional well-being. However, even though, these professionals value the interpersonal relationship they establish with their patients, their main concern is technical. Patients, on the other hand, link nursing care to the manifestation of attention, affection and patience. The hermeneutic analysis revealed that nursing care is based on a human predisposition for assistance. This assistance can either be committed and conscious, when it is concerned and solicitous, or uncompromising when it is an inauthentic occupation which only meets mundane immediatisms. We concluded that this study promotes the consolidation of comprehensive-philosophical investigations which can contribute to the qualification of the nursing care.

Adult↗