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Biomedical subjects

D Pepin

Publications and source records attributed to D Pepin.

31 records · Page 2Linked to original sources

Essential fatty acid pattern of glycerolipids in rat hepatocytes in primary culture and in coculture with rat liver epithelial cells.

The linoleic acid content of phosphatidylethanolamine (PE), phosphatidylcholine (PC) and triglyceride (TG) rapidly fell in rat hepatocytes in primary culture up to four days and in coculture with liver epithelial cells up to eight days. At the same time, the level of polyunsaturated fatty acids (PUFA), especially arachidonic acid, remained constant in PE, slightly decreased in PC and dropped in TG. There was no variation of the nonessential PUFA, 20:3n-9. Linoleic acid supplementation of cultures 24 hr before the harvest induced a rise in the linoleic acid level of the three lipid classes. Arachidonic acid remained constant in TG and only slightly decreased in PE and PC at day 4 of primary culture and day 8 of coculture. The level of 20:3n-9 increased in PE and PC and much more in TG. This net increase in the arachidonic acid and 20:3n-9 levels in TG could not be explained only by a transfer from the phospholipid pools of PUFA because the phospholipid content of hepatocytes and PUFA levels of phospholipids did not vary under linoleic supplementation. The low percentage of arachidonic acid in epithelial cells rules out any participation of these cells in the increase of arachidonic acid in supplemented cocultures. Triglycerides may act as a storage pool for plasma PUFA up to four days of primary culture and eight days of coculture. Besides, coculture seems more potent than primary culture to maintain the phospholipid level, to spare the essential PUFA in PE and to increase the TG synthesis in response to linoleic acid supplementation.

Animals↗

Antigenic relatedness between phospholipases A2 from Naja naja venom and from mammalian cells.

Polyclonal antiserum was prepared against phospholipase A2 from Naja naja and used to prepare a purified antibody. It cross-reacted with the antigen, and with intracellular mammalian PLA2. This antibody was immunoreactive and inhibited the PLA2 activity of Naja naja and of guinea pig alveolar macrophages or rat lymphocytes. By immunoblotting, this antiserum revealed one band of PLA2 from Naja naja (14 kDa) and 3 bands for guinea pig alveolar macrophages and rat lymphocytes (30, 45 kDa and a minor band of 14 kDa). These results show an antigenic relatedness between an extracellular PLA2 and membrane-bound PLA2 from two different mammalian species and cell types.

Animals↗

[Thermodynamic studies of ontogenic rhythms of energy exchange in the alyte (an anuran amphibian)].

The total energy flux in an Amphibian was measured using a microcalorimetric technique throughout different stages of development. Simultaneous records of the individual activity (actography) allowed us to study circadian rhythms and their variations during ontogenesis. Acrophase of the energy exchanges take place at the beginning of the night in adults, tadpoles and juveniles of more than 9 months. However, there was a shift in the acrophase of the circadian rhythm in juveniles aged between 7 and 9 months. This could be an adaptation process which regulate competition within populations of the same species.

Aging↗

Essential fatty acid uptake and esterification in primary culture of rat hepatocytes.

Primary cultures of adult rat hepatocytes were used to compare the uptake and esterification of essential polyunsaturated fatty acids (18:2, 20:3 and 20:4 of the n-6 series) with those of palmitic and oleic acids. The uptake of unesterified fatty acids was linearly related to the free fatty acid/albumin molar ratio for 14 h and did not depend on the unbound free fatty acid level. Whatever the initial free fatty acid/albumin molar ratio, it dropped to 0.5 +/- 0.1 mM after 14 h, thus showing that hepatocytes have a high capacity for clearing free fatty acids from the medium at high free fatty acid/albumin molar ratios. The free fatty acid uptake become saturable when the free fatty acid and albumin concentrations were raised and the free fatty acid/albumin ratio remained constant. This strongly suggests that albumin-hepatocyte interaction mediates free fatty acid uptake. This uptake was identical whatever the fatty acid tested and did not depend on the relative amounts of fatty acids when they were added simultaneously. Triacylglycerol accumulation and synthesis, monitored by labelled fatty acids, were related to the free fatty acid/albumin molar ratio and exhibited no specificity for the series of fatty acids tested. Triacylglycerols were enriched in all the fatty acids tested by up to 60%, and fatty acid incorporation into diacylglycerols and triacylglycerols reflected the free fatty acid composition of the medium. By contrast, neither the level nor the synthesis of phospholipids varied with free fatty acid/albumin, but the rate of phospholipid turnover depended on the fatty acids tested. Accumulation of these acids was smaller in phospholipids than in triacylglycerols. When linoleic and arachidonic acids were added together, phospholipids (especially phosphatidylethanolamine and phosphatidylinositol) were more enriched in arachidonic acid than triacylglycerols. This might be due to the specificity for fatty acid of the enzymes involved in phospholipid metabolism.

Animals↗

Essential fatty acid interconversion during gestation in the rat.

The synthesis of arachidonic acid has been investigated in fetal and pregnant rat liver microsomes in the course of the gestation. The delta 5-desaturase activity decreased 2-3 times in rat liver between the 19th and 22nd day of the pregnancy. During this period the delta 5-desaturate activity increased 3-fold in the fetal liver, exceeding the activity of the maternal liver. In contrast, the activity of the fetal delta 6-desaturase was in the same range as in pregnant rat liver and the liver of control animals and did not change between these two stages of the gestation. The elongation rate of linoleic acid in fetal liver was 2-3 times lower than in maternal liver but this increased during the pregnancy. The fatty acid activate rate was always higher than the activity of the desaturases. At the 19th day, the activity of the delta 5-desaturase was apparently the rate limiting step of arachidonic acid synthesis in fetal liver. We did not find any delta 5- and delta 6-desaturase activities or linoleic acid elongation in the placenta microsomes.

Animals↗

Essential fatty acids interconversion in the human fetal liver.

In order to study the role played by the fetoplacental unit in providing the human fetus with arachidonic acid, delta 5- and delta 6-desaturase activities were studied in microsomes from human fetal liver and placenta after 18 and 22 weeks of gestation. We evidenced for the first time delta 5- and delta 6-desaturase activities in fetal liver microsomes. As in adult liver, delta 6-desaturation is the rate-limiting step of arachidonic acid synthesis. No activity was found in the placenta. Arachidonic acid concentrations were higher in fetal serum than in maternal serum while the opposite was observed for linoleic acid. The fetal liver microsomal content in arachidonic acid was low. Taken together the data suggest that arachidonic acid is supplied to the fetus through a preferential transfer across the placenta.

Chromatography, Gas↗

Protein-stimulated enrichment of human platelet membranes in linoleylphosphatidylcholines. Effect upon adenylate cyclase and fluidity.

In order to study the effect of linoleyl enrichment of platelet membranes upon adenylate cyclase activity and membrane fluidity, manipulations of platelet phospholipids are carried out with phosphatidylcholine-loaded high-density lipoproteins (HDL) or phospholipid-exchange protein and phospholipid-cholesterol mixed vesicles. Incubation with HDL does not appear to be valuable for this purpose. On the other hand, phospholipid-exchange protein and mixed vesicles can be used successfully. Phospholipid-exchange protein stimulated 3-fold the spontaneous exchange of 2-linoleylphosphatidylcholine between the vesicles and the platelets. Linoleyl enrichment of platelets by dilinoleylphosphatidylcholine is about 25% and by 2-linoleylphosphatidylcholine is about 45-50%. The unsaturation index remains constant when the enrichment is performed using dilinoleylphosphatidylcholine but it increases with 2-linoleylphosphatidylcholine. Basal and prostaglandin E1-stimulated adenylate cyclase activities are not modified by dilinoleylphosphatidylcholine, while they increase significantly in the case of 2-linoleylphosphatidylcholine. There is no significant variation in diphenyl hexatriene fluorescence polarization parameters, either with dilinoleylphosphatidylcholine or with 2-linoleylphosphatidylcholine.

Adenylyl Cyclases↗

[Platelet arachidonic acid and central retinal vein obstruction (author's transl)].

Study of 25 cases of obstruction of the central retinal vein. In the 4 first days it is a low level of arachidonic acid of platelet membrane. This fall of level is the one anomaly of fatty acid from platelet and plasma. We have obtained no correlation between the level of platelet AA and age or lipids level of the blood. Study of platelet agregation was performed in only some cases and no conclusion is possible.

Adult↗

[Plasma bile acids fractionnation by gas-liquid chromatography : application to hepatocellular deficiency investigation (author's transl)].

The level of the four major bile acids was measured in the plasma using gas liquid chromatography. The enhancement of the level of the primary bile acids was well known during bile duct obstruction. In the case of hepatic diseases, the chenodesoxycholiccholic acid concentration ratio (CDC/C) was equal or lower than unity when cholestasis occurred rather than hepatic deficiency. When hepatic deficiency developed, the ratio CDC/C was higher than unity. Plasma bile acid fractionnation exhibits also a prognostic value. When the ratio CDC/C is very high, it is significant of progressive hepatic deficiency. A high level of lithocholic acid is also a sign of unfavourable prognosis.

Bile Acids and Salts↗

Chemical and in vitro toxicological evaluations of water packaged in polyvinyl chloride and polyethylene terephthalate bottles.

This study proposed a new strategy assessing the health risks of mineral water packaging and compared the chemical analytical techniques and some in vitro cytotoxicological assays for the study of PVC and PET materials at the main stages of the manufacturing process of bottles. These evaluations were carried out with food simulant (deionized-endotoxin-tested water) and with natural mineral water in real conditions of packaging and storage (from 0 to 24 months). The complementarity of these two approaches is discussed. Some analytical and cytotoxic abnormalities were detected in the food simulant after contact with the batches of powdered PVC compound, PET resin and their intermediate steps of transformation (PVC-'paraison', PET-'perform'). But these results did not reflect the actual behaviour of the finished PVC and PET bottles, for which no major abnormality was detected in the natural mineral water.

Animals↗

Plasma and erythrocyte essential fatty acids during total parenteral nutrition in infants: effects of a cutaneous supply.

In order to prevent essential fatty acid (EFA) deficiency induced by fat-free total parenteral nutrition (TPN), 10 infants on TPN were rubbed three times daily for 20 days using oenethera oil (80% EFA). Total EFA amount provided cutaneously was 1900 mg/kg/d. Plasma and red blood cells phospholipids were determined on days 1 and 20 in these 10 treated and six untreated infants on TPN and compared with those of normal control infants. On day 1, plasma nonessential FA including 20:3 n-9(p less than 0.01) were increased in both TPN groups while 18:2 n-6 and 18:3 n-3 (p less than 0.001 and p less than 0.01) were decreased. On the 20th day, EFA deficiency had worsened with a decrease in plasma level of 20:4 n-6 (p less than 0.02) and a higher than normal triene/tetraene ratio : 3.4 +/- 1.1 and 2.3 +/- 0.6 vs 0.1 +/- 0.1 (p less than 0.02). As for red blood cells phospholipids, 16:0 was increased and 18:2 n-6 and 20:3 n-6 were decreased (p less than 0.05) on day 1. On day 20, these FA were more abnormal while 20:3 n-9 became significantly increased (p less than 0.05). No difference was observed between the TPN groups at any time. These results show that cutaneous application of large amounts of EFA-rich oil is unable to prevent or cure TPN induced EFA deficiency.

Absorption↗