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Biomedical subjects

L Vigh

Publications and source records attributed to L Vigh.

At least 37 records · Page 2Linked to original sources
PubMed1990

Action of a homogeneous hydrogenation catalyst on living Tetrahymena mimbres cells.

Various conditions were tested in an attempt to hydrogenate the unsaturated fatty acids of living Tetrahymena mimbres with the homogeneous catalyst palladium di-(sodium alizarine monosulfonate) without causing serious damage to the cells. Using a low (20 micrograms/ml) catalyst concentration in the external medium, hydrogenation of greater than 20% of surface membrane lipid double bonds were obtained, but hydrogenation of intracellular membranes was minimal. When exposed to H2, cells preincubated with inactive catalyst for several hours and visibly loaded with the catalyst lost viability as soon as hydrogenation exceeded trace levels. Material secreted by Tetrahymena into their medium effectively inhibited hydrogenation of added oleic acid, normally a good substrate. Mucus secreted by the cells, soluble proteins isolated from cell homogenates, bovine serum albumin, and cysteine were also inhibitory, but the inhibition could be overcome by employing higher catalyst concentrations. Although some enzymatic retroconversion of saturated lipids back to unsaturated lipids appeared to take place, the scale of the conversion was small, and further experimentation will be required to understand the mechanism involved. The selective hydrogenation of surface membranes achieved by these methods may be especially useful to those interested in fluidity effects on plasma membrane properties.

Animals↗
PubMed1989

Effect of catalytic hydrogenation of Tetrahymena ciliary phospholipid fatty acids on ciliary phospholipase A activity.

Detached Tetrahymena cilia were treated for increasing periods of time with the homogeneous hydrogenation catalyst palladium di(sodium alizarine monosulphonate). This caused a 4-70% reduction in the number of double bonds in phospholipid-bound fatty acids and a concurrent decrease in membrane fluidity as detected by ESR measurements. Ciliary phospholipase A activity was markedly inhibited when as little as 13% of the fatty acid double bonds had been hydrogenated, suggesting that the enzyme activity is very sensitive to changes in membrane fluidity.

Animals↗
PubMed1988

Recovery of Dunaliella salina cells following hydrogenation of lipids in specific membranes by a homogeneous palladium catalyst.

Unsaturated fatty acyl chains of Dunaliella salina membrane lipids can be catalytically reduced by the homogeneous hydrogenation catalyst palladium di(sodium alizarine monosulphonate), Pd(QS)2, under conditions permitting full recovery of the cells within 24 h. The hydrogenation is accomplished by incubation of cells with the hydride form of Pd(QS)2 under 1 atmosphere of H2 and for 2 min or less. Following this protocol, hydrogenation reduces only those fatty acids located in the plasma membrane and other membranes located near the cell surface. The limited reactivity in vivo is due to the fact the Pd(QS)2 permeates into the living cells more slowly than it does into liposomes prepared from extracted Dunaliella membrane lipids. While Dunaliella is completely unaffected by exposure to the oxygenated, inactive catalyst, hydrogenated cells cease growth for approximately 12 h, during which time the hydrogenated acyl chains are being enzymatically retroconverted to their original unsaturated form. When the lipid composition approaches its prehydrogenation values, growth resumes, presumably due to the restoration of normal membrane functions. The system shows promise for studying the metabolic regulation of membrane microviscosity.

Chlorophyta↗
PubMed1988

Lipid Saturation Induced Microviscosity Increase Has No Effect on the Reducibility of Flash-Oxidized Cytochrome f in Pea Thylakoids.

Homogeneous catalytic hydrogenation was used to modify the level of fatty acid unsaturation of thylakoid membranes in the pea chloroplast. Fluidity alteration has been monitored simultaneously using the spin-label probe, 16-doxyl stearate. Even in the case of 30% hydrogenation, no change in the reduction rate of flash-oxidized cytochrome f was observed, in contrast to the fact that the same decrease in the double-bond content of the thylakoid membrane resulted in a pronounced inhibition in the full-chain electron transport. We conclude that the rate of lateral diffusion of reduced plastoquinone is unaffected by the lowering of the fluidity of the thylakoid lipid matrix.

Journal Article↗
PubMed1987

The hydrogenation of phospholipid-bound unsaturated fatty acids by a homogeneous, water-soluble, palladium catalyst.

The hydrogenation of unsaturated phospholipids by palladium di(sodium alizarine monosulphonate) activated for 5 min under H2 proceeded rapidly at 20 degrees C and 1 atm. H2. Multibilayer liposomes of dioleoyl- and dilinolenoylphosphatidylcholine were hydrogenated at similar rates while dilinoleoyl- and 1-palmitoyl-2-oleoylphosphatidylcholine were hydrogenated at slightly slower rates. The reduction of polyunsaturated fatty acids gave rise to a variety of natural and unnatural positional cis and trans isomers which were largely reduced further to saturated fatty acids as the hydrogenation continued. Dioleoylphosphatidylethanolamine was attacked by the catalyst more slowly at 20 degrees C than was the equivalent phosphatidylcholine molecular species. Experiments conducted using mixtures of phosphatidylethanolamine and phosphatidylcholine in varying proportions also suggested that phospholipids are slightly more susceptible to catalytic hydrogenation in the bilayer phase than in the hexagonalII phase. Understanding the sequence of hydrogenation reactions involving these one and two component lipid preparations is useful in interpreting the action of the palladium catalyst on living cells under the same mild conditions.

Catalysis↗
PubMed1987

Nitrate starvation induces homeoviscous regulation of lipids in the cell envelope of the blue-green alga, Anacystis nidulans.

Replacement of the normal culture liquid to a nitrate-free medium resulted in an immediate drop in the ratio of protein to lipid in isolated cell envelopes of Anacystis nidulans cells. The relative fluidity of the envelope membranes or liposomes, made from the extracted lipids of the envelope, was estimated by measuring the steady-state fluorescence polarization of 1,6-diphenyl-1,3,5-hexatriene. A thermotrophic phase transition of lipids within the cytoplasmic membrane of intact cells was also revealed by detecting the temperature-dependent absorption changes in the proportion of zeaxanthin at 390 nm. It became evident that a decrease in the proportion of protein to lipid within the cell envelope was accompanied neither by changes in the microviscosity level, nor by shifting of characteristic temperatures of the liquid-crystalline-to-gel transition of lipids. In parallel with nitrate starvation, however, the proportion of saturated fatty acids of the envelope lipids increased markedly. Accumulation of saturated, longer-chain (C18) fatty acids at the cost of C16 counterparts upon nitrate deprivation occurred in all of the complex lipids. In accordance with these findings, a pronounced decrease in the fluidity was demonstrated for the liposomes prepared from the envelope polar lipids of nitrate-starved cells compared with the corresponding control, throughout the temperature range (45-5 degrees C) studied. We propose that the fluidizing effect due to a fall in the ratio of protein to lipid was compensated by a rapidly triggering regulatory process which enables the preservation of the fluidity characteristics at an optimal level within the cell envelope of A. nidulans.

Cell Membrane↗
PubMed1987

Catalytic hydrogenation of fatty acyl chains in plasma membranes; effect on membrane lipid fluidity and expression of cell surface antigens.

Optimal reaction conditions were established for hydrogenation of plasma membranes of living murine GRSL leukemia cells, using the water-soluble catalyst Pd(QS)2 (QS, sulphonated alizarine; C14H6O7NaS). Under these conditions more than 80% of the cells remained viable. Analysis by gas chromatography revealed that hydrogenation occurred predominantly in the 18:2, 20:4 and 22:6 fatty acyl chains of the membrane phospholipids. Under the same conditions hydrogenation was also performed in purified plasma membranes from GRSL cells and from rat liver, and in liposomes prepared from the total lipid extracts of these membranes. Hydrogenation increased the lipid structural order parameter in the membranes, as measured by fluorescence polarization. This increase was more pronounced in the liposomes (46%) than in the plasma membranes (17-25%). Hydrogenation increased the expression of a 15 kDa antigen on the surface of viable GRSL cells, as measured in a Fluorescence Activated Cell Sorter, using monoclonal antibodies. The expression of four other antigens, among which H-2k, was not or much less affected by this treatment.

Animals↗
PubMed1986

Effects of ascorbic acid in vivo on the fatty acid composition of the tissues of mice treated with Gramoxone.

The effects of the simultaneous administration of ascorbic acid and the LD50 of paraquat (an ingredient of Gramoxone), and of ascorbic acid pretreatment followed by the LD50 of paraquat, were studied on the phospholipid and triglyceride fatty acid levels in homogenates of mouse lung, liver and kidney. Ascorbic acid treatment increases the content of polyunsaturated fatty acids in the lung considerably, i.e. the pulmonary membrane fluidity decreases significantly in response to ascorbic acid. In the liver homogenate, the membrane fluidity is significantly increased by ascorbic acid pretreatment, and significantly decreased by simultaneous ascorbic acid treatment. In the renal tissue, the result of ascorbic acid pretreatment exhibits a similar tendency to that of paraquat treatment, but a more significant one, while the administration of ascorbic acid together with paraquat does not cause a substantial change in the fluidity index compared to the control.

Animals↗
PubMed1986

Primary Role of the Cytoplasmic Membrane in Thermal Acclimation Evidenced in Nitrate-Starved Cells of the Blue-Green Alga, Anacystis nidulans.

The lipid phase transition of the cytoplasmic membrane and the chilling susceptibility were studied in nitrate-starved Anacystis nidulans cells. Nitrate starvation resulted in the disappearance of the thylakoid membrane system, without any effect on chilling susceptibility. The chilling susceptibility of the algal cells depended on the growth temperature. Temperatures of lipid phase transitions of the cytoplasmic membranes were detected by chilling-induced spectral changes in the carotenoid region, in vivo. These values were identical to those of cultures containing intact thylakoid systems. Our results suggest that cytoplasmic membrane plays a determinative role in the thermal acclimation of the alga cells.

Journal Article↗
PubMed1985

Modulation of chloroplast membrane lipids by homogeneous catalytic hydrogenation.

A method is reported for the modification of lipids in situ in chloroplast membrane by which a homogeneous, water-soluble catalyst Pd(QS)2 (QS, sulphonated alizarine; C14H6O7NaS) is incorporated into the thylakoids of isolated chloroplast. The catalyst itself did not affect the photosynthetic activity but caused an extensive loss of unsaturated fatty acids in the presence of hydrogen gas. The polyunsaturated fatty acids were hydrogenated at a faster rate than the monoenoic acids. During hydrogenation the orientational ordering of membrane lipids, as measured with the C-12 positional isomer of spin-labelled stearic acid, displayed a slight increase in agreement with the alterations in membrane composition. Progressive saturation of double bonds of lipids primarily inhibits electron transport between the photosystems followed by the inhibition of electron flow around photosystem II. Photosystem I electron transport was not inhibited even by 50% fatty acid hydrogenation. We suggest that using Pd(QS)2 catalyst for thylakoid hydrogenation offers an excellent technique to study the role of various unsaturated fatty acids in the regulation of membrane fluidity and photosynthetic processes.

Catalysis↗
PubMed1985

Modulation of membrane fluidity in living protoplasts of Nicotiana plumbaginifolia by catalytic hydrogenation.

A homogeneous water-soluble Ru catalyst, has been incorporated into mesophyll protoplasts isolated from Nicotiana plumbaginifolia leaves. In the presence of hydrogen gas this complex causes an extensive loss of unsaturated fatty acid bonds and a concomitant increase in microviscosity of the cellular membranes. Although the gradual reduction of the level of unsaturation, per se, is accompanied by considerable cell damage, there is an optimum reaction time where approximately 50% of the protoplasts are still living and about 20% of the double bounds initially present in fatty acyl residues have undergone hydrogenation. The possible mechanism of the self regulatory process competing with the hydrogenation in the early stages of the reaction is also discussed.

Catalysis↗
PubMed1985

Effect of frost hardening on lipid and Fatty Acid composition of chloroplast thylakoid membranes in two wheat varieties of contrasting hardiness.

Lipid and fatty acid composition of chloroplast thylakoid membranes was determined in two varieties of wheat (Triticum aestivum L.), the hardy Miranovskaja and the sensitive Penjamo. Plants were grown at room temperature or under frost hardening conditions (1.5 degrees C). Changes in lipid and fatty acid composition of the isolated thylakoids could be related to the temperature dependence of light-stimulated proton uptake. Changes in the thylakoid phospholipids upon hardening of the two varieties did not show any direct relation with low temperature tolerance of light-dependent H(+) uptake; neither did changes in phospholipid fatty acid chain lengthening to 20 and 22 C-atoms in combination with increased desaturation up to 6 double bonds. Increased low temperature tolerance of light-induced H(+) uptake by hardening was correlated with the following glycolipid changes: maintained glycolipid level, a proportionally increased digalactosyl diglyceride fraction, a decrease in thylakoid monogalactosyl diglyceride, increased sulfolipid fatty acid chain lengthening (20 and 22 C-atoms), and increased sulfolipid desaturation (4-6 double bonds). We suggest that the above mentioned changes in glycolipids have adaptive value for low temperature tolerance of light-dependent proton uptake.

Journal Article↗
PubMed1983

Changes induced by Gramoxon in tissue phospholipids and phospholipid fatty acids in mouse and guinea-pig.

Following administration of the LD50 or the LD100 of Gramoxon (PQ), the phospholipids (PL) of the lung, liver and kidney were separated and both the PLs and the fatty acids isolable from them were examined quantitatively. The different doses of PQ caused different changes of the saturated and unsaturated fatty acids in the various organs. The changes in the PL unsaturated fatty acids point to PQ-induced lipid peroxidation enhancement and membrane damage.

Animals↗
PubMed1982

Isolation of a wheat cell line with altered membrane properties.

A spontaneous dimethylsulfoxide (DMSO)-tolerant cell line was isolated from a cell culture of wheat (Triticum monococcum L.). The tolerant cells were able to grow in the presence of 4% DMSO. Cells formed from protoplasts of the tolerant line required DMSO for division in culture medium of high osmotic value.Fatty acid composition and the molar ratio of phospholipids/sterols suggest a more ordered membrane structure in the tolerant line. Accordingly, a lower K(+) influx rate was detected in the tolerant cells in comparison with the original line. These characteristics were maintained after 6 months' cultivation of the cells in DMSO-free growth medium. This suggested that genetic changes could be responsible for differences between the two cell lines.

Journal Article↗
PubMed1980

Relation or Raman order parameters to spin labeling parameters.

For the quantitation of Raman and spin labeling data order parameters are commonly used. The spin label order parameter measured at any depth in the layer is a weighed sum of the segmental order since, due to fast conformational interconversions, each CH2 segment is partly in trans and partly in non-trans, e.g. gauche, kink, jog, etc. conformation during the measurement. The weighing factor, the trans finding probability, varies along the chain (cf. flexibility profile) but its mean value should be equal to the Raman trans order parameter. This correlation is illustrated with the experimental data obtained for dipalmitoyl phosphatidylcholine and n-alcohol mixtures. The rate of rotational diffusion, a dynamical parameter from spin labeling studies, is correlated with the lateral packing density as measured by the Raman lateral order parameter. For the obtained linear correlation a qualitative explanation is given. The effect of a series of long chain alcohols on the phase transition characteristics of dipalmitoyl phosphatidylcholine was investigated. The possible role of hydrogen bonding in the interfacial region is emphasized.

Alcohols↗