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Biomedical subjects

P Garcia-Barreno

Publications and source records attributed to P Garcia-Barreno.

At least 19 recordsLinked to original sources

Fluorescent and compositional changes in crystallin supramolecular structures in pig lens during development.

Water soluble proteins (WSPs) in Sus scrofa lenses from pigs in different developmental stages: (young (GI), young adult (GII), and middle-aged (GIII)) were separated using GF-HPLC, yielding fractions of different molecular weights. Non-tryptophan (345/420 nm) and tryptophan (280/345 nm) fluorescence was measured in these fractions. Relative non-tryptophan fluorescence increased with age at a rate directly correlated to the molecular weight of aggregates forming the different chromatographic fractions, while tryptophan fluorescence tended to decrease. The crystallins constituting each fraction were separated using 2D-electrophoresis and after development with Coomassie blue they were identified using MS-TOF. Also, the protein content of each spot was quantified by subsequent scanning and integration. The proportions of unchanged crystallins characteristically changed with age in chromatographic fractions of different molecular weights. Thus it was possible to relate these changes with those occurring in the fluorescent properties and molecular weight of supramolecular structures.

Age Factors↗

Development and clinical assay of the BCM ventricular assist device.

The development of a ventricular assist device is depicted. The seven subprograms that make up the project are described briefly. Results obtained during the clinical assay showed that the new design of the input cannula working as a false auricle provides better efficiency, as well as a certain level of autoregulation for the device.

Aged↗

Association of changes in lysophosphatidylcholine metabolism and in microsomal membrane lipid composition to the pulmonary injury induced by oleic acid.

Alterations in the lipid composition of lung microsomal membranes occur in oleic acid-induced respiratory distress. The marked decrease in the phosphatidylcholine/lysophosphatidylcholine molar ratio could be related with an altered metabolism of lysophosphatidylcholine in these membranes. Results revealed that the activity of phospholipase A increased whereas that of acyl-CoA:lysophosphatidylcholine acyltransferase decreased. Microsomal lysophospholipase activity remained unchanged. On the other hand, the microsomal enzyme system involved in the de novo synthesis of diacylglycerol was impaired, and cholinephosphotransferase activity was lowered. These changes in the activity of some membrane-bound enzymes were not caused by changes in the membrane lipid fluidity since lipid structural order parameter (SDPH) did not change and neither did the major factors on which the fluidity depends. The possible significance of microsomal lipid alterations in the pathogenesis of respiratory distress induced by oleic acid is discussed.

1-Acylglycerophosphocholine O-Acyltransferase↗

Normal biochemistry values in baboons (Papio C. cinocephalus).

1. Analysis of biochemical parameters was carried out on material pooled from 10 baboons, adult male and female, of the Papio c. cinocephalus strain. 2. The values determined were for the common metabolites and enzymes utilized for specific studies in general metabolism (urea, glucose, cholesterol, phospholipids, bilirubin, uric acid, creatinine, alkaline phosphatase, glutamic oxaloacetic transaminase, glutamic pyruvic transaminase, lactic dehydrogenase, leucine amino peptidase, gamma glutamyl transpeptidase, total creatine kinase and amylase). Total protein and its fractions (ratio albumin/globulin, albumin, alpha 1-globulin and alpha 2-globulin) were also determined. 3. Results of these studies were compared with values for normal human adults. 4. The differences obtained between the respective ranges can be explained by the different physiology of the two species and, possibly, may also be caused by the use of human substrates in the enzyme assays. 5. Through the determination of normal values for the baboon, this study provides values for this species as an experimental animal in biomedical research.

Alkaline Phosphatase↗

Amniotic fluid catecholamine metabolites in maternal smoking.

The concentration of dopamine (DA), norepinephrine (NE), and its metabolites 3-methoxy-4-hydroxy-phenylethylene glycol (MHPG) and vanillyl mandelic acid (VMA), and prostaglandins E2, and F2a in amniotic fluid were measured at labor (1-3 cm of cervical dilatation), in uncomplicated and complicated pregnancies. The amniotic fluid concentration (mean +/- SD) of MHPG in maternal smoking (S) (30.6 +/- 11.9 nmol.mg creatinine-1) and intrauterine growth retardation (IUGR) (36.7 +/- 15.2 nmol.mg creatinine-1) groups is significantly higher than in uncomplicated pregnancies (C) (22.9 +/- 8.8 nmol.mg creatinine-1), or diabetes mellitus groups (D) (26.1 +/- 10.9 nmol.mg creatinine-1) groups; VMA presents similar differences between groups S (145.8 +/- 84.4 nmol.mg creatinine-1) and IUGR (173.0 +/- 135.4 nmol.mg creatinine-1) higher than C (58.3 +/- 26.1 nmol.mg creatinine-1) and D (76.8 +/- 49.0 nmol.mg creatinine-1) groups. NE levels differ between groups in a similar way, but DA does not present significant differences between groups. Prostaglandin levels are higher in the S than in the C group. Nicotine action and/or hypoxia are proposed as a possible cause of NE, MHPG, and VMA rise in amniotic fluid of S, and IUGR versus C or D groups.

Amniotic Fluid↗

Comparison between intra- and extracellular surfactant in respiratory distress induced by oleic acid.

The present study compares the phospholipid distribution and protein content in bronchoalveolar lavage, purified extracellular surfactant and lamellar bodies isolated from rabbits killed at intervals of 2.5, 12 and 24 h after oleic acid administration. The data suggest that the alteration of pulmonary surfactant could be partially due to the type II cell response to the injury.

Animals↗

Microsomal membrane fluidity and phosphatidylcholine synthesis in rabbit lung under high oxygen tension.

Phosphatidylcholine metabolism and membrane fluidity were studied in microsomes isolated from rabbit lung, which had been exposed to high oxygen tension for 30 min. In these microsomes the incorporation of [3H]-palmitate into phosphatidylcholine increased whereas the incorporation of [14C]-glycerol and [14C]-choline from CDP-[methyl-14C]-choline remained unchanged in comparison to the control microsomes. The enhanced [3H]-palmitate incorporation may be explained by an increase of the specific activity of acyl-CoA:lysophosphatidylcholine acyltransferase which was measured in microsomes from hyperoxic lung. Although microsomal parameters influencing membrane fluidity, such as the cholesterol/phospholipid molar ratio, unsaturation degree of phospholipid acyl chains and lipid/protein ratio, are altered after oxygen treatment in vivo, no change of fluorescence polarization (PDPH) and lipid structural order parameter (SDPH) could be measured. Probably, the membrane maintains its fluidity by counteracting effects on different factors on which the fluidity depends.

Acylation↗

Lipid composition of miniature pig platelets.

1. Analyses of platelet lipid composition were carried out on material pooled from male and female miniature pigs. 2. The cholesterol/phospholipid molar ratio was 0.6. 3. Phosphatidylcholine represents the major class of phospholipids (47%) and phosphatidylinositol the minor (2%). 4. The main fatty acids of phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol and sphingomyelin were: palmitic, stearic, oleic, linoleic and arachidonic acids. 5. The ratios of saturated to unsaturated fatty acids were: sphingomyelin, 1.7; phosphatidylcholine, 1.2; phosphatidylserine, 0.9; phosphatidylethanolamine and phosphatidylinositol, 0.6. 6. Our results suggests that human and miniature pig platelet lipids bear several characteristics in common. This fact would allow miniature pig to be used as a new experimental model.

Animals↗

Rapid stimulation of diacylglycerol production in Xenopus oocytes by microinjection of H-ras p21.

The p21 products of ras proto-oncogenes are thought to be important components in pathways regulating normal cell proliferation and differentiation. These proteins acquire transforming properties as a result of activating lesions that convert ras genes to oncogenes in a wide array of malignancies. In Xenopus laevis oocytes, microinjection of transforming ras p21 is a potent inducer of maturation, whereas microinjection of a monoclonal antibody to ras p21 inhibits normal maturation induced by hormones. The phosphoinositide pathway is a ubiquitous system that appears to play a key role in diverse cellular functions. By use of the Xenopus oocyte system, it was possible to quantitate the effects of ras p21 microinjection on individual components of the phosphoinositide pathway. Within 20 minutes of microinjection, levels of phosphatidylinositol 4,5-bisphosphate, inositol 1-phosphate, and inositol bisphosphate increased 1.5- to 2-fold. The most striking effects were on diacylglycerol, which increased 5-fold under the same conditions. In contrast, the normal ras p21 protein induced no detectable alteration in any of the metabolites analyzed. The earliest effects of the transforming p21 on phosphoinositol turnover were observable within 2 minutes, implying a very rapid effect of ras p21 on the enzymes involved in phospholipid metabolism.

Animals↗

Zymosan-induced release of inositol phosphates at resting cytosolic Ca2+ concentrations in macrophages.

Hydrolysis of polyphosphoinositides by phosphodiesterase has been demonstrated to be involved in the control of cytosolic Ca2+ concentrations. The stimulation of Ca2+ ionophores of the release of inositol phosphates in macrophages, and other cells, together with the Ca2+ requirements for zymosan-induced phospholipase C activation, make unclear the relationship between Ca2+ mobilization and polyphosphoinositide hydrolysis. The results in the present paper strongly suggest that, for zymosan-induced phospholipase C activation, a previous increase in cytosolic Ca2+ is not a required event. These results also show that zymosan-activated release of inositol phosphates may be mediated by a guanine-nucleotide-binding protein.

Animals↗

Effect of ethanol on the arachidonic acid metabolism in mouse peritoneal macrophages.

Macrophages play an important role in the development of chronic inflammatory states. Ethanol has been shown to impair a number of membrane-linked phenomena. The synthesis and secretion of oxygenated metabolites of arachidonic acid is triggered at the cytoplasma membrane level. The present study was carried out in order to investigate the effect of ethanol on the arachidonic acid metabolism in mouse peritoneal macrophages. Two types of experiments were performed: with endogenous radiolabeled arachidonic acid and with exogenously added radiolabeled arachidonic acid. Our data show that ethanol in vitro activates the release of arachidonic acid from intracellular pools, while the proportion of endogenous substrate metabolized in the presence of ethanol is similar to that in controls. From the exogenous it seems clear that ethanol induces different effects depending whether the arachidonic acid is endogenous or added exogenously.

Animals↗

Effects of Escherichia coli lipopolysaccharide on the phosphoinositide metabolism and serotonin secretion in thrombin-activated platelets.

Lipopolysaccharide treatment of platelets in basal conditions does not produce any effect on serotonin secretion or on phosphatidic acid synthesis or arachidonic acid release. However, a brief exposure of platelets to endotoxin enhances the thrombin-stimulated activation of these parameters, whereas a more prolonged treatment with lipopolysaccharide impairs thrombin action. The results presented here also suggest that very short-term treatment of platelets with lipopolysaccharide activates the inositol 1,4,5-trisphosphate 3-kinase. The long-term effects of endotoxin could be mediated by protein kinase C activation.

Animals↗

Phospholipase C-diglyceride lipase is a major pathway for arachidonic acid release in macrophages.

Macrophages are a rich source of arachidonic acid oxygenated metabolites and play a remarkable role in a number of physiopathological situations. The synthesis and secretion of arachidonic acid metabolites are triggered at the cytoplasmic membrane level. The present study was outlined to further investigate the cellular mechanisms controlling arachidonic acid release in macrophages. The results presented here strongly suggest that the amount of arachidonic acid released in macrophages in response to phagocytic challenge could be accounted for by a phospholipase C-diglyceride lipase system being unnecessary the presence of phospholipase A2 whose activity, on the other hand, was found vanishingly small in macrophage homogenates.

Animals↗

Arachidonic acid releasing systems in pig aorta endothelial cells.

Endothelial cells synthesize prostacyclin both from platelet-derived endoperoxides and from the arachidonic acid released from its intracellular stores. The mechanisms controlling this release does not appear to be mediated through phospholipid methylation but by means of phosphoinositide hydrolysis. As yet two possible mechanisms have so far been proposed to regulate arachidonic acid release in a number of cellular systems: phospholipase C-controlled phospholipase A2 activity or phospholipase C-diglyceride lipase system. The results presented here show that using phospholipases inhibitors is not a reliable strategy to study arachidonic acid release in cultures of endothelial cells. Our data also strongly suggest that the release of prostacyclin may be accounted in these cells for by a phospholipase C-diglyceride lipase system.

Animals↗

Ionophore A23187 induces a refractory state in thrombin-activated release of inositol phosphates.

The phosphatidylinositol cycle has been proposed to be involved in the regulation of platelet functionality through the control of cytoplasmic Ca2+ levels. However, the requirements of phospholipase C for Ca2+ has not yet been elucidated in intact platelets. The primary purpose of the present study was to investigate the Ca2+ requirements of this enzyme in platelets from miniature swine by taking advantage of the permeabilizing properties of the ionophore A23187. Our results strongly suggest that the treatment of platelets with A23187 induces a refractory state in thrombin-stimulated release of inositol phosphates while 5-hydroxytryptamine (serotonin)-secretory capacity in response to thrombin remained constant. This refractory state seems to be dependent on some cytochalasin-inhibitable cytoskeletal phenomena.

Animals↗

The microviscosity of liver plasma membranes of rats fed with oleoylanilide.

Oleoylanilide was administered orally to groups of rats according to different patterns. Oleoylanilide was perfused at different concentrations through rat liver. Oleoylanilide was added to isolated hepatocytes. Oleoylanilide was added to plasma-membrane preparations. Membrane preparations were obtained after experiments performed in vivo and perfusion experiments and, by using 1,6-diphenylhexa-1,3,5-triene as fluorescence probe, the fluorescence polarization parameter was measured, from which the microviscosity (eta) was calculated. In all cases the microviscosity decreased markedly. Addition of oleoylanilide to hepatocyte preparations and to isolated membranes produced the same effect, increasing the fluidity of the membranes. These data suggest that oleoylanilide partitions into the membrane, disordering some lipid interactions.

Anilides↗

Lipogenesis in liver, lung and adipose tissue of rats fed with oleoylanilide.

Oleoylanilide was administered orally to groups of rats according to different patterns. Subcellular fractionation of liver, lung and adipose tissue was then carried out in order to study the main enzyme activities involved in the lipogenesis. The observed findings indicate that adipose tissue and lung are the main target organs for the anilide, adipose tissue being involved in a general decrease of the enzyme activities, whereas transacylation reaction exhibits the most marked depletion of all the enzyme activities in the lung. The enzyme activities in liver were not markedly affected by this oral administration, although some data support the existence of a latent liver toxicity. These data suggest that oleoylanilide has the capacity to alter lipid metabolism of lung and adipose tissue to a considerable extent, whereas no major effect was produced in the liver. This different organ response could be related to the lymphatic gland via absorption of the substance.

Adipose Tissue↗