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C Alves

Publications and source records attributed to C Alves.

47 records · Page 3Linked to original sources
PubMed1994

[Why do endothelial cells require adenosine triphosphate?].

NMR-spectroscopy (31P-NMR, 23Na-NMR) has been used to characterize the energy metabolism of cultured endothelial cells of the pig aorta and to determine the energy requirements of the Na-K-ATPase relative to total cellular energy consumption. Endothelial cells exhibited high concentrations of creatine phosphate and cardiac microvascular creatine kinase (CK) was of the BB-CK subtype. BB-CK can therefore be used as a marker-enzyme of endothelial cells in muscular organs. The transmembrane flux-rate of sodium was 117 mumol Nalmin/mg prot in the steady state at an intracellular sodium concentration of 25 mmol/l. Combining microcalorimetric measurements with 23Na-NMR data revealed that the energy requirement of endothelial Na-K-ATPase despite the high surface to volume ratio comprises only 3-5% of total cellular energy consumption.

Adenosine Triphosphate↗
PubMed1993

Bile acid transport into hepatocyte smooth endoplasmic reticulum vesicles is mediated by microsomal epoxide hydrolase, a membrane protein exhibiting two distinct topological orientations.

Bile acids, such as taurocholate, have been shown to be transported into hepatocyte smooth endoplasmic reticulum (SER) vesicles. This process is Na(+)-independent, electrogenic, inhibitable by 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid and taurochenodeoxycholate, with a Km of 352 microM and a Vmax of 29.6 nmol/mg protein/min. The observed transport is mediated by the bifunctional protein, microsomal epoxide hydrolase (mEH) which can also mediate bile acid transport into hepatocytes across the sinusoidal plasma membrane (von Dippe, P., Amoui, M., Alves, C., and Levy, D. (1993) Am. J. Physiol. 264, G528-G534). mEH was isolated from SER membranes by immunoprecipitation with monoclonal antibody (mAb) 25D-1 which recognizes this protein on the surface of intact hepatocytes. The SER-derived protein exhibited an apparent molecular weight, isoelectric point, N-terminal amino acid sequence, and mEH-specific activity that were indistinguishable from the plasma membrane form of the enzyme. Proteoliposome reconstitution of the SER taurocholate transport system indicated that mEH was absolutely required for the expression of transport capacity. The interaction of mAb 25D-1 with mEH on intact right-side-out SER vesicles demonstrated that the epitope found on the surface of hepatocytes was also found on the cytoplasmic surface of these vesicles (80%) and in the lumen (20%) suggesting the presence of two forms of this protein in the SER, the latter from being sorted to the cell surface. The existence of two orientations of this protein in the SER was confirmed by the sensitivity to tryptic digestion, where 75% of the mAb epitope was accessible to the enzyme. The loss of the 25D-1 epitope correlated with loss of taurocholate transport capacity. The role of mEH in the transport process and the orientation of the transporting isoform was further established by demonstrating that mAb 25A-3, which also reacts with mEH on the hepatocyte surface, was able to directly inhibit taurocholate transport in the SER vesicle system. These and previous results thus establish that isoforms of mEH can mediate taurocholate transport at the sinusoidal plasma membrane and in SER vesicles and that this bifunctional protein can exist in two orientations in the SER membrane. The association of bile acids with the SER suggests a possible role of intracellular vesicles in the transhepatocellular movement of bile acids from the sinusoidal to the canalicular compartment.

Animals↗
PubMed1993

Na(+)-dependent bile acid transport by hepatocytes is mediated by a protein similar to microsomal epoxide hydrolase.

A protein mediating hepatocyte sodium-dependent bile acid transport across the sinusoidal plasma membrane has been purified by immunoprecipitation with monoclonal antibody (MAb) 25D-1, which specifically recognizes this protein on the surface of intact hepatocytes (Ananthanarayanan et al. J. Biol. Chem. 263: 8338-8343, 1988). The function of this protein was further established by proteoliposome reconstitution (von Dippe et al. J. Biol. Chem. 265: 14812-14816, 1990). NH2-terminal amino acid sequence analysis and amino acid composition revealed this protein to be closely related to the enzyme microsomal epoxide hydrolase (mEH). Both proteins exhibited the same elution times on a reverse-phase high-pressure liquid chromatography column, comigrated with an apparent molecular weight of 49,000 as measured by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and possessed identical isoelectric points of 8.2. The MAb was capable of immunoprecipitating chromatographically purified mEH, as well as a protein derived from the sinusoidal plasma membrane that exhibited mEH activity comparable to that of the protein isolated from the endoplasmic reticulum. The subtilisin fragmentation patterns derived from chromatographically purified mEH and the MAb-precipitated plasma membrane protein were also identical. Hydropathy profile analysis of the amino acid sequence of mEH suggested the presence of four transmembrane domains. The results of these studies indicate that a protein that is involved in mediating sodium-dependent bile acid transport is closely related to mEH.

Animals↗
PubMed1990

Effects of hypoxia and fatty acids on the distribution of metabolites in rat heart.

The effects of exogenous fatty acids and hypoxia on cardiac energy metabolism were studied by measuring mitochondrial and cytosolic adenine nucleotides as well as CoA and carnitine esters using a tissue fractionation technique in non-aqueous solvents. During normoxia, the administration of 0.5 mM palmitate caused a considerable increase in acyl-CoA and acylcarnitine, particularly in mitochondria. High-energy phosphates, however, were only slightly altered. A 90 min low-flow hypoxia caused a dramatic increase in mitochondrial acyl esters. The mitochondrial ATP content decreased significantly, while the cytosolic concentration was only slightly diminished, suggesting an inhibition of mitochondrial adenine nucleotide translocation by long-chain acyl-CoA. Addition of palmitate during hypoxia amplified hypoxic damage and reduced adenine nucleotides in both compartments considerably, while fatty acid metabolites were only slightly affected. In presence of an inhibitor of fatty acid oxidation (BM 42.304), the fatty-acid-induced acceleration of cardiac injury was prevented. Since BM 42.304 decreased mitochondrial acylcarnitine and increased the cytosolic concentration significantly, BM 42.304 was presumed to inhibit mitochondrial acylcarnitine translocase. However, a causal relationship between lipid metabolites and ischemic damage seemed unlikely.

Acyl Coenzyme A↗
PubMed1990

Height screening in the community. The commercialization of growth. The role of the pediatrician.

The availability of unlimited but costly supplies of biosynthetic growth hormone has led to pressure for pharmacologic use (as opposed to replacement therapy in proven deficiency states). Commercial and altruistic motives have converged to promote community height screening among individuals who have been perceived by themselves or parents as short. This does not meet accepted criteria for health screening. Height screening of large populations of children yields few unrecognized medical conditions. If the goal of community screening is to identify abnormally short individuals (less than 3%) who might benefit from growth hormone treatment and if the unproven assumption is correct that stature correlates with success and happiness, then those less likely to appear for screening need to be recruited to avoid elitist domination. The annual cost of such growth promotion would be greater than $10 billion, with no evidence for substantial health benefits. Growth monitoring of all children through improved height measurement in schools and in physicians offices, as part of health supervision, is a more sound community approach than height screening.

Body Height↗
PubMed1989

Graves disease presenting as painful thyroiditis.

Hyperthyroidism associated with subacute (painful, viral) thyroiditis is well-recognized as a clinical entity; the thyroid gland in Graves disease is minimally, if ever, tender and painful. We describe a 10-year-old girl with hyperthyroidism whose initial clinical presentation was predominantly a painful, tender goiter. Graves disease was established by high uptake of 131I with a diffuse pattern of distribution of radioactivity on scan and the presence of thyroid-stimulating antibody. Thyrotropin-binding inhibiting IgG and antibody to thyroid microsomal antigen were both positive. She responded well to treatment with propylthiouracil and had spontaneous regression of her thyroid pain. The cause of the severe pain and tenderness remains speculative.

Child↗
PubMed1988

Lysolecithins as endothelium-dependent vascular smooth muscle relaxants that differ from endothelium-derived relaxing factor (nitric oxide)

The effects of lysolecithin (lysophosphatidylcholine) derived from egg yolk as well as of synthetic lysolecithins with different aliphatic chain lengths on tension development of rabbit aortic strips were investigated. Lysolecithins caused slowly progressing, dose-dependent relaxation that was inhibited by hemoglobin, methylene blue, and nordihydroguiaretic acid. Indomethacin caused no inhibition of relaxation. The degree of relaxation was endothelium-dependent and appeared to be related to the activation of guanylate cyclase [GTP pyrophosphate-lyase (cyclizing), EC 4.6.1.2]. Superoxide dismutase failed to influence relaxation. Lysolecithins with the longest aliphatic chain were the most potent relaxants of aortic strips. The experiments suggest a role of lysolecithins through their weak detergent action on membrane dynamics of endothelial cells, resulting in the production of cyclic GMP and the relaxation of arterial smooth muscle. Lysolecithins differ in several respects from endothelium-derived relaxing factor. Endothelium-derived relaxing factor is an unstable humoral substance released from endothelium and is identical to nitric oxide, itself a labile substance causing vascular relaxation and cyclic GMP accumulation. Lysolecithins may represent a different type of endothelium-dependent muscle relaxant.

Acetylcholine↗
PubMedDate not supplied

Skin reaction in Callithrix jacchus penicillata to phytohemagglutinin.

Sixty marmosets (Callithrix jacchus penicillata) were intradermally tested with phytohemagglutinin. It was possible to classify this population into three groups on the basis of the reactions: 1. negatives, 2. those which showed just local hemorrhage, and finally 3. a group which developed a wheal as well as hemorrhage. The biopsy at the site of the reaction demonstrated several microscopic foci. This finding contrasted with the homogeneous cellular infiltration found in the human controls.

Animals↗