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Acute response of rat liver microsomal lipids, lipid peroxidation, and membrane anisotropy to a single oral dose of polybrominated biphenyls.

Liver microsomal lipids and lipid peroxidation activities were examined in adult male rats at intervals over a 2-mo period after the administration of a single oral dose of 0 or 500 mg/kg of FireMaster BP-6 in corn oil. Microsomal lipids were markedly altered in the polybrominated biphenyl- (PBB-) dosed animals at the earliest time examined (1 wk), and these changes persisted throughout the remainder of the study. An early decrease in the cholesterol-phospholipid ratio was noted, which probably contributed to the significant decrease in the steady-state fluorescence anisotropy demonstrable in both intact microsomes and in liposomes prepared from microsomal lipid extracts. Significant concentrations of PBBs were present in dosed rat microsomes, but the changes in anisotropy appeared to result from membrane lipid alterations rather than from a direct perturbation by PBBs. Iron ascorbate-induced peroxidation was also greatly enhanced in dosed rat microsomes, even when rats were maintained on a low-iron (25 ppm) diet. These early alterations in membrane fluidity and peroxidative capacity of microsomes may ultimately contribute to the hepatotoxicity of PBBs.

Administration, Oral↗

The pathway of membrane fusion catalyzed by influenza hemagglutinin: restriction of lipids, hemifusion, and lipidic fusion pore formation.

The mechanism of bilayer unification in biological fusion is unclear. We reversibly arrested hemagglutinin (HA)-mediated cell-cell fusion right before fusion pore opening. A low-pH conformation of HA was required to form this intermediate and to ensure fusion beyond it. We present evidence indicating that outer monolayers of the fusing membranes were merged and continuous in this intermediate, but HA restricted lipid mixing. Depending on the surface density of HA and the membrane lipid composition, this restricted hemifusion intermediate either transformed into a fusion pore or expanded into an unrestricted hemifusion, without pores but with unrestricted lipid mixing. Our results suggest that restriction of lipid flux by a ring of activated HA is necessary for successful fusion, during which a lipidic fusion pore develops in a local and transient hemifusion diaphragm.

Cells, Cultured↗

In vivo hemoglobin dosimetry of malonaldehyde and ethene in mice after induction of lipid peroxidation. Effects of membrane lipid fatty acid composition.

Hemoglobin (Hb) adduct determination by the N-alkyl Edman method was used for in vivo dosimetry of endogenously formed malonaldehyde (MA) and ethene in mice fed diets with different fatty acid composition and induced for lipid peroxidation with carbon tetrachloride (CCl4). In order to amplify lipid peroxidation animals were pretreated with phenobarbital (PB) and the glutathione-depleting agent DL-buthionine-(S,R)-sulfoximine (BSO). Non-treated animals raised on different diets were used as controls. Lipid peroxidation products in liver were measured as 2-thiobarbituric acid reactive compounds (TBA-C). Livers from control mice fed a soya oil based diet (rich in polyunsaturated fatty acids, diet S) showed approximately 6.5-fold higher levels of TBA-C than those from animals raised on a coconut oil based diet (mostly saturated fatty acids, diet C). The level of adducts of MA to Hb, determined as N-(3-hydroxypropyl)valine, was approximately 1.5-fold higher in animals from diet S than in animals raised on diet C. The highest increases in the levels of TBA-C and MA adducts were obtained after a simultaneous treatment of the animals with PB, BSO and CCl4. The increases of TBA-C were 1.3-fold (diet C) and 1.7-fold (diet S). The corresponding increases of MA-Hb adduct levels were 1.3- and 1.6-fold respectively, indicating an increased susceptibility of mice fed diet S to lipid peroxidation. The level of adducts from ethene, determined as N-(2-hydroxyethyl)valine, was also higher in mice from diet S than in animals fed diet C, when all treatment groups were considered. The difference was, however, only slightly significant (P less than 0.02). No difference between control and CCl4-treated animals, with regard to the ethene-Hb adduct, was found. Our results validate the use of Hb dosimetry for monitoring the effects of factors known to influence lipid peroxidation induced in vivo.

Animals↗

Effect of dietary level of ascorbic acid on the growth, hepatic lipid peroxidation, and serum lipids in guinea pigs fed polychlorinated biphenyls.

Rats exposed to polychlorinated biphenyls (PCB) or other xenobiotics exhibit an increase in tissue and urinary ascorbic acid, serum cholesterol and hepatic lipid peroxidation. To clarify the physiological role of ascorbic acid in exposure to PCB, we studied the influence of dietary levels of ascorbic acid (30-2,000 ppm) on the growth, serum lipids and hepatic lipid peroxidation in guinea pigs fed 50 ppm PCB-containing diets. The results showed that the growth depression due to PCB was ameliorated by increasing dietary ascorbic acid. The increases in serum cholesterol and phospholipid and in hepatic lipid peroxidation due to PCB were suppressed by larger amounts of ascorbic acid, which inversely correlated with the changes in growth. PCB intake also increased serum levels of cortisol and triglyceride, but these effects were not influenced by dietary level of ascorbic acid. The optimum requirement of ascorbic acid in the guinea pigs fed PCB was 800-2,000 ppm in the diet for the changes in the growth, serum cholesterol and hepatic lipid peroxidation.

Animals↗

Soluble non-starch polysaccharides derived from complex food matrices do not increase average lipid droplet size during gastric lipid emulsification in rats.

The creation of a finely dispersed lipid emulsion is essential for efficient hydrolysis of dietary triglycerides. The effectiveness of emulsification within the stomach depends upon the shear force generated by gastric motility and the concentration of emulsifiers present in the gastric contents. Other dietary constituents can modify these factors, and previous studies have suggested that the presence of soluble non-starch polysaccharides (NSP) during digestion might increase the average size of intraluminal emulsion droplets. In the present study, we developed a new technique for the isolation and analysis of intraluminal lipid emulsions by optical diffraction analysis. The method was applied to rats fed powdered semipurified diets that were free of all NSP or supplemented with insoluble cellulose, guar gum, or NSP derived from apple, carrot or rolled oats. Cellulose had no significant effect on emulsion size, and there was no evidence that the average sizes of lipid droplets in the gastric fundus or antrum were higher than control values in rats fed diets supplemented with any source of soluble NSP. In the groups fed oats and cooked carrot NSP, the mean droplet diameters approached half the values for diets free of NSP or containing insoluble cellulose. The difference between rats fed NSP from cooked carrot and those fed cellulose was significant in the proximal stomach (P < 0.05), and that between rats fed raw oats and rats fed cellulose was significant in the distal stomach (P < 0.05). Soluble dietary fiber does not inhibit lipid or cholesterol absorption via any inhibition of lipid emulsification.

Absorption↗

Inverse regulation of lipid-peroxidizing and hydroperoxyl lipid-reducing enzymes by interleukins 4 and 13.

12/15-lipoxygenases and phospholipid hydroperoxide glutathione peroxidases are opposite enzymes balancing the intracellular concentration of hydroperoxy lipids. We studied the regulation of both enzymes by interleukins 4 and 13 and found an inverse response. When human lung carcinoma cells A549 were cultured in vitro in the presence of these cytokines, an up-regulation of the 12/15-lipoxygenase and a down-regulation of the phospholipid hydroperoxide glutathione peroxidase were observed. A similar inverse regulation was found in human peripheral monocytes. Interleukin 4-treated A549 cells exhibited an impaired capability of reducing exogenous hydroperoxyl lipids and their levels of endogenous lipid hydroperoxides were markedly increased. To find out whether these regulatory processes also occur in vivo, arachidonic acid oxygenase and phospholipid hydroperoxide glutathione peroxidase activity was assayed in various tissues of transgenic mice that systemically overexpress interleukin 4. In lung, spleen, kidney, and heart, an increased arachidonic acid oxygenase activity was detected when transgenic mice were compared with inbred controls. The phospholipid hydroperoxide glutathione peroxidase activity was impaired in lung, liver, and spleen of the transgenic animals. These data indicate that lipid-peroxidizing and lipid peroxide-reducing enzymes are inversely regulated in various mammalian cells. Up-regulation of the 12/15-lipoxygenase and simultaneous down-regulation of the phospholipid hydroperoxide glutathione peroxidase may lead to an increased oxidizing potential, which is reflected by an augmented intracellular peroxide tone.

Animals↗

Structure and synthesis of a lipid-containing bacteriophage. Effects of lipids containing cis or trans fatty acids on the reconstitution of bacteriophage PM2.

Infectious PM2 virus paticles could be reconstituted in vitro from a mixture of nucleocapsid, phospholipids containing cis fatty acids, and proteins I and II. The presence or absence of acyl phosphatidylglycerol, a minor lipid component of thevirion, did not affect the reconstitution of infectious particles, even though it was incorporated into the particles when present. When phosphatidylglycerol was completely replaced by acyl phosphatidylglycerol in the reconstitution mixture, no infections particles were formed. Lipids containing either cis or trans fatty acids were also used for reconstitution in vitro of the lipid-containing bacteriophage PM2. Regardless of the ratio of phosphatidlyglycerol to phospatidylethanolamine in the reconstitution mixture, infectious particles were formed and had almost the same phospholipid composition when lipids containing cis-palmitoleic acid were used; no infectious particles were obtained when lipids containing trans-palmitoleic acid were used. In the latter case, virus-like particles were, however, formed. Reconstituted particles containing cis fatty acids were infectious when tested on wild type Pseudomonas BAL-31 as well as on the unsaturated fatty acid auxotroph grown in the presence of either cis or trans-palmitoleic acid. Reconstituted particles containing trans fatty acids were not infectious on any of these cells. When trans fatty acids as well as cis fatty acids were present in the reconstitution mixture, then there was a lower yield of infectious particles. Particles with either cis or trans fatty acids had all four viral proteins and adsorbed to BAL-31 host cells in a specific manner.

Bacteriophages↗

Organophosphate metasystox-induced increment of lipase activity and lipid peroxidation in cerebral hemisphere: diminution of lipids in discrete areas of the rat brain.

The effects of the pesticide, metasystox (O,O-dimethyl-S-2 or Metasystor (ethylsulphinyl) ethylthiophosphate), on various lipid fractions in the discrete areas of the brain were studied. The daily intraperitoneal administration of 4 mg/kg body weight of metasystox for 10 days has depleted the levels of total lipids, phospholipids, cholesterol, esterified fatty acids and gangliosides in cerebral hemisphere, cerebellum, brain stem and spinal cord. But the lipase and lipid peroxidation measured in the cerebral hemisphere were significantly increased. It is possible that the inhibition of the lipid levels in the discrete areas of the brain may be due to the increase activity of lipase and lipid peroxidation.

Animals↗

Lipid metabolism in Achlya: studies of lipid turnover during development.

The life cycle of Achlya involves germination of spores to form coenocytic somatic hyphae, followed by differentiation of hyphal tips into sporangia. From germination to release of new spores occupies 27-30 h. Total lipid made up 10% of dry weight in ungerminated spores. After germination, total lipid fell to 6% of dry weight in 15 h, then rose to 7.7% at the time of sporangium formation. Half of the initial loss of lipid took place within 2 h of germination. The ability of Achlya to incorporate [1-14C]acetate into lipid was maximal at the time of sporangium formation, and glycerides were the principal component of total lipid to become 14C-labelled at all stages of the life cycle. Fatty acid synthetase activity measured in cell-free extracts was low in spores and in actively elongating mycelium, but increased during idfferentiation to a level 15-fold greater than that in spores. Fatty acid oxidation, as estimated by the release of 14CO2 from 1-14C-labelled fatty acids, was also maximal at the time of sporangium formation.

Acetates↗

On the role of lipid-bilayer elasticity for the lipid-protein interaction and the indirect protein-protein coupling.

The present paper deals with the curvature (or splay) elasticity of lipid bilayers and its possible consequences for the microscopic organization of membranes. Experimental studies of the microstructure of lipid-lipid and lipid-protein mixtures are reported and discussed in terms of this type of membrane elasticity. In particular, evidence is provided that the elastic distortion of lipid bilayers caused by the incorporation of proteins can lead to mechanisms of the indirect protein-protein interaction. Finally the question is discussed whether these mechanisms could also play a role for the formation of enzyme complexes in biological membranes.

Dimyristoylphosphatidylcholine↗

The effects of cytomegalovirus infection on polar lipids and neutral lipids in cultured human cells.

The effects of infection by the human cytomegaloviruses Ad-169 on the incorporation of [14C]acetate into the polar and neutral lipids of human embryonic lung cells and human saphenous vein smooth muscle cells were compared to [14C]acetate incorporation in mock-infected control cells. Cytomegalovirus infection caused a shift in the relative amounts of polar and neutral lipids, with infected cells having lower amounts of polar lipids and higher amounts of neutral lipids than mock-infected controls. When neutral lipids were separated into diglyceride (DG), cholesterol (C), fatty acid, triglyceride (TG) and cholesterol ester (CE) components, Ad-169-infected cells had lower levels of incorporation of label into CE, TG, and DG fractions, and higher levels of label incorporation into C than mock-infected cells.

Acetates↗

Lipid composition, physical state, and lipid peroxidation of tumor membranes.

Studies were carried out on microsomes isolated from the highly differentiated (slow-growing) Morris hepatoma 9618A, on microsomes and plasma membranes from the poorly differentiated (fast-growing) Morris hepatoma 3924A, and rat liver used as control. The lipid composition (phospholipid and cholesterol content, degree of fatty acid unsaturation) and peroxidation of such membranes has been correlated with the order and fluidity of the membrane bilayer. The results indicate that substrate availability is the rate-limiting step in microsomal and plasma membrane lipid peroxidation of hepatoma 3924A. From diphenylhexatriene fluorescence depolarization measurements it appears that the changes in lipid composition cause an increase in the order of the lipid bilayer on going from the control to hepatoma 9618A and 3924A microsomes, while fluidity is virtually unchanged. Conversely, for similar chemical changes, in plasma membranes from hepatoma 3924A the order is nearly the same and there is a decrease in fluidity. The changes in the above parameters of tumor membranes might be partly related to the loss of protective enzymes against oxygen radicals. This is supported by the observation that inhibition of liver superoxide dismutase and glutathione reductase, by treatment of rats with diethyldithiocarbamate and chloroethyl nitrosourea, respectively, renders the microsomal membranes more resistant to lipid peroxidation in vitro.

Adenosine Diphosphate↗

Plasma lipases and lipid transfer proteins increase phospholipid but not free cholesterol transfer from lipid emulsion to high density lipoproteins.

BACKGROUND: Plasma lipases and lipid transfer proteins are involved in the generation and speciation of high density lipoproteins. In this study we have examined the influence of plasma lipases and lipid transfer protein activities on the transfer of free cholesterol (FC) and phospholipids (PL) from lipid emulsion to human, rat and mouse lipoproteins. The effect of the lipases was verified by incubation of labeled (3H-FC,14C-PL) triglyceride rich emulsion with human plasma (control, post-heparin and post-heparin plus lipase inhibitor), rat plasma (control and post-heparin) and by the injection of the labeled lipid emulsion into control and heparinized functionally hepatectomized rats. RESULTS: In vitro, the lipase enriched plasma stimulated significantly the transfer of 14C-PL from emulsion to high density lipoprotein (p<0.001) but did not modify the transfer of 3H-FC. In hepatectomized rats, heparin stimulation of intravascular lipolysis increased the plasma removal of 14C-PL and the amount of 14C-PL found in the low density lipoprotein density fraction but not in the high density lipoprotein density fraction. The in vitro and in vivo experiments showed that free cholesterol and phospholipids were transferred from lipid emulsion to plasma lipoproteins independently from each other. The incubation of human plasma, control and control plus monoclonal antibody anti-cholesteryl ester transfer protein (CETP), with 14C-PL emulsion showed that CETP increases 14C-PL transfer to human HDL, since its partial inhibition by the anti-CETP antibody reduced significantly the 14C-PL transfer (p<0.05). However, comparing the nontransgenic (no CETP activity) with the CETP transgenic mouse plasma, no effect of CETP on the 14C-PL distribution in mice lipoproteins was observed. CONCLUSIONS: It is concluded that: 1-intravascular lipases stimulate phospholipid transfer protein mediated phospholipid transfer, but not free cholesterol, from triglyceride rich particles to human high density lipoproteins and rat low density lipoproteins and high density lipoproteins; 2-free cholesterol and phospholipids are transferred from triglyceride rich particles to plasma lipoproteins by distinct mechanisms, and 3 - CETP also contributes to phospholipid transfer activity in human plasma but not in transgenic mice plasma, a species which has high levels of the specific phospholipid transfer protein activity.

Animals↗

Lipid metabolism in rats fed diets containing different types of lipids.

OBJECTIVE: To assess the effect of different types of lipid diets on the lipid metabolism of aging rats. METHODS: Fifty male Wistar rats were studied from the time of weaning to 12 and 18 months of age. Their diets were supplemented as follows: with soybean oil (S), canola oil (CA), lard and egg yolk (LE), and canola oil + lard and egg yolk (CA + LE). Blood pressure (BP) was measured every month, and the heart/body ratio (H/BR) was determined. The rats were euthanized at the age of 12 and 18 months, and blood samples were collected for lipid analysis as follows: total cholesterol (TC), LDL-C, VLDL-C, HDL-C, triglycerides (TG), and glucose. RESULTS: The type of oil ingested by the animals significantly altered BP, H/BR, and serum lipid levels in rats at 12 and 18 months. No difference was observed in the survival curve of the animals in the different groups. The LE group had the highest BP, and the CA group was the only one in which BP did not change with aging. A reduction in the H/BR was observed in the LE and CA+LE animals. At the age of 12 months, differences in TC, HDL-C, LDL-C, VLDL-C, TG, and glucose were observed. At the age of 18 months, a significant difference in TC, HDL-C, and glucose was observed. The highest TC value was found in the CA group and the lowest in the S group. CONCLUSION: No increase in BP occurred, and an improvement was evident in the lipid profile of rats fed a diet supplemented with CA, in which an elevation in HDL-C levels was observed, as compared with levels with the other types of diet.

Aging↗

Effects of vitamin E-enriched egg yolk on lipid peroxidation, hemolysis and serum lipid concentration in young and old rats.

Effects of a diet containing different amount of vitamin E-enriched egg yolk powder on lipid peroxidation, hemolysis, and serum lipid concentration were examined in young and old rats. Young and old rats were fed experimental diets containing 5.6% or 22.5% of normal egg yolk powder, and 5.6% or 22.5% of vitamin E-enriched egg yolk powder. When young rats were fed these diets, their tocopherol concentrations in testes, heart, kidneys, and retroperitoneum fat pads increased according to the diet tocopherol contents. Lipid peroxide levels in these tissues were inversely proportional to the tocopherol concentrations. The liver and serum, however, did not show the above relationship. In the rats fed vitamin E-enriched egg yolk diets hemolysis did not increase, in contrast to those fed normal egg yolk diets. The rats fed a 5.6% vitamin E-enriched egg yolk diet had a lower serum cholesterol level than those fed a normal egg yolk diet. In old rats, one control group was used to determine initial tocopherol concentration and two other groups were fed 5.6% normal and vitamin E-enriched egg yolk diets. The vitamin E-enriched egg yolk rats showed lower weights of the body, liver, and retroperitoneum fat pads at the end of the experiment. The normal egg yolk group had atrophied testes. The tissue and serum tocopherol concentrations were increased in the vitamin E-enriched egg yolk group, whereas they were decreased in the normal egg yolk group. In the vitamin E-enriched egg yolk group, lipid peroxide, hemolysis and serum cholesterol, especially HDL-cholesterol, did not increase. These results indicate that egg yolk tocopherol was absorbed and prevented lipid peroxidation and hemolysis in young and old rats, and testes atrophy in old rats.

Age Factors↗

Role of lipids, lipoproteins and lipid peroxidation in thrombocytopenia in patients with vivax malaria.

Possible roles of oxidative and metabolic changes of lipids in the development of thrombocytopenia in patients with vivax malaria were investigated. Serum total cholesterol, HDL and LDL cholesterols, triglycerides, lipid peroxidation level and platelet counts were determined in 60 patients with vivax malaria and compared with 50 healthy individuals. Inter-relationships of parameters were investigated. In patients, serum total cholesterol, HDL and LDL cholesterol concentrations and platelet counts were lower, and serum triglycerides and plasma lipid peroxidation concentrations were higher than those of control subjects. Significant relationships were found among serum lipid parameters, plasma lipid peroxidation and thrombocyte counts. Decrease in total cholesterol, HDL and LDL cholesterols and platelet counts may be related or secondary to oxidative stress. In the treatment of thrombocytopenia required in the some cases of malaria, the supplementation of antioxidative vitamins may be considered.

Adolescent↗

[Fliformine effects on blood lipid spectrum and lipid peroxidation in obese patients].

Blood lipid spectrum and lipid peroxidation are compared in patients with different obesity types. Diet treatment is compared to biguanides derivatives (gliformine) in patients with body obesity combined with dislipidemia. Gliformine proved more effective in correction of changed lipid spectrum of atherogenic origin, improves lipid peroxidation. Biguanides seem perspective in management of lipid disbolism in patients with body obesity.

Biguanides↗

Lipid-protein stoichiometries in a crystalline biological membrane: NMR quantitative analysis of the lipid extract of the purple membrane.

The lipid/protein stoichiometries of a naturally crystalline biological membrane, the purple membrane (PM) of Halobacterium salinarum, have been obtained by a combination of (31)P- and (1)H-NMR analyses of the lipid extract. In total, 10 lipid molecules per retinal were found to be present in the PM lipid extract: 2-3 molecules of phosphatidylglycerophosphate methyl ester (PGP-Me), 3 of glycolipid sulfate, 1 of phosphatidylglycerol, 1 of archaeal glycocardiolipin (GlyC), 2 of squalene plus minor amounts of phosphatidylglycerosulfate (PGS) and bisphosphatidylglycerol (archaeal cardiolipin) (BPG) and a negligible amount of vitamin MK8. The novel data of the present study are necessary to identify the lipids in the electron density map, and to shed light on the structural relationships of the lipid and protein components of the PM.

Archaeal Proteins↗