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Biomedical subjects

H Sies

Publications and source records attributed to H Sies.

At least 289 records · Page 16Linked to original sources
PubMed1987

Alcohol, aldehydes and lipid peroxidation: current notions.

The relations between membrane damage, lipid peroxidation and the effects of alcohol have attracted longstanding interest. In vitro studies showed that parameters of lipid peroxidation were positive. However, in vivo studies did not always provide clearcut conclusions. Our interest and that of a number of other groups, therefore, has centered on the intact organ and intact animal, using noninvasive indicators of lipid peroxidation such as ethane or pentane release and low-level chemiluminescence. The studies were mainly carried out with the isolated perfused rat liver, and there is substantial evidence for aldehydes being crucial in promoting lipid peroxidation. The range of aldehydes was extended from acetaldehyde to longer-chain aldehydes; in particular, aldehydes which themselves can originate from polyunsaturated fatty acid breakdown, e.g. 4-hydroxynonenal and congeners, were studied and were found to be potent in supporting lipid peroxidation.

Aldehydes↗
PubMed1986

Mutagenicity of nitroxide-free radicals.

Stable nitroxides were found to be mutagenic using Salmonella typhimurium tester strain TA 104, a strain chosen on the basis of its high sensitivity to oxidative damage. Nitroxide mutagenicity was dramatically increased in the presence of the superoxide radical generating system, xanthine oxidase/hypoxanthine, and it was suppressed in cells carrying the oxyR1 mutation, which causes induction of enzymes protecting against oxidative stress. As nitroxide-free radicals occur biologically, e.g., in the metabolism of aromatic amines, these radical-induced mutations could be a model for the carcinogenicity observed with these compounds.

Cyclic N-Oxides↗
PubMed1986

The effect of urea synthesis on extracellular pH in isolated perfused rat liver.

In a non-recirculating system of isolated liver perfusion, stimulation of urea synthesis by NH4Cl is followed by a decrease of effluent pH by up to 0.2 pH unit. This effect is not observed when urea synthesis is inhibited by amino-oxyacetate or norvaline. When the urea formed by the liver is immediately hydrolysed with urease before the effluent perfusate reaches the pH electrode, the urea-synthesis-induced acidification is no longer observed. This indicates that accompanying alterations in hepatic metabolism after stimulation of urea synthesis, such as increased energy provision and consumption, are not responsible for the extracellular acidification, but that the effect is due to the formation of urea itself. The acidification of the extracellular space after stimulation of urea synthesis by NH4Cl is quantitatively explained by the consumption of 2 mol of HCO3-/mol of urea formed: 1 mol being incorporated into urea, the other being protonated to yield CO2 and H2O. The data match the theoretically predicted HCO3- consumption during ureogenesis and underline the role of hepatic urea synthesis for disposal of HCO3- by converting it into the excretable products CO2 and urea.

Ammonium Chloride↗
PubMed1986

Energy-linked cardiac transport system for glutathione disulfide.

The relationship between the rate of glutathione disulfide (GSSG) export and the energy state was studied in isolated perfused rat heart. The intracellular GSSG level was maintained at saturation for transport (7.5 nmol GSSG X min-1 X g heart-1) by continuous perfusion with 20 microM t-butyl hydroperoxide. GSSG release was substantially restricted upon the addition of inhibitors of mitochondrial respiration such as KCN, antimycin A or rotenone. In contrast, no effect was observed on GSSG release during potassium-induced cardiac arrest, although changes in oxygen consumption and coronary flow were similar to those observed with KCN. The dependence of the GSSG transport rate on the cytosolic free ATP/ADP ratio reveals that GSSG transport is half-maximal at (ATP/ADP)free approximately equal to 10. The capacity of GSSG transport was unchanged by infusion of epinephrine, norepinephrine or dibutyryl cyclic AMP.

Adenine Nucleotides↗
PubMed1986

Role of cardiac glutathione transferase and of the glutathione S-conjugate export system in biotransformation of 4-hydroxynonenal in the heart.

There is a remarkable difference in the isozyme pattern between cardiac and hepatic glutathione S-transferases in rat (Ishikawa, T., and Sies, H. (1984) FEBS Lett. 169, 156-160), and one near-neutral isozyme (pI = 6.9) of the cardiac glutathione S-transferases was found to have a significantly high activity toward 4-hydroxynonenal. The isozyme was inhibited by the resulting glutathione S-conjugate of 4-hydroxynonenal competitively with GSH and noncompetitively with 4-hydroxynonenal. The kinetic parameters estimated for the isozyme were: kcat = 460 mol X min-1 X mol enzyme-1, Km = 50 microM for 4-hydroxynonenal, Ki = 85 microM. When the heart was perfused with 4-hydroxynonenal, a marked decrease was observed in the intracellular GSH level, accompanied by an increase of glutathione S-conjugate of 4-hydroxynonenal in the heart. The rate of the conjugation reaction was more than 30 times the rate of the spontaneous reaction, the half-life of 4-hydroxynonenal in the heart being less than 4 s. The glutathione S-conjugate of 4-hydroxynonenal was released from the heart into the perfusion medium. Saturation kinetics were observed for the release with respect to the intracellular level of the S-conjugate (Vmax = 12 nmol X min-1 X g heart-1), and there was a competition by the S-conjugate for GSSG release. The release of the glutathione S-conjugate is considered as a carrier-mediated process and to be important not only in interorgan glutathione metabolism but also in diminishing the inhibitory effect of the S-conjugate on glutathione S-transferases and glutathione reductase.

Aldehydes↗
PubMed1986

The major isozyme of rat cardiac glutathione transferases. Its correspondence to hepatic transferase X.

A major isozyme of rat heart glutathione transferase was purified to homogeneity by Sephadex G-200 gel filtration, ammonium sulfate precipitation, CM-cellulose chromatography and affinity chromatography on S-hexylglutathione-linked Sepharose 6B. The purified isozyme was a dimer with an apparent relative molecular mass of 50 000 composed of two Yb-size subunits (Mr = 26 500). The isozyme is immunologically related to rat liver glutathione transferase X and 3-3, especially closely to transferase X, and no immunological cross-reactivity with subunits 1 and 2 of hepatic glutathione transferases was observed. The isoelectric point (pI = 6.9) of the isozyme was identical with and the substrate specificity was very similar to transferase X. Thus, the cardiac near-neutral isozyme is considered to be identical to glutathione transferase X recognized in rat liver. The amount of this near-neutral isozyme estimated to be present in heart tissue is 70 micrograms/g. The isozyme has relatively high activities towards alpha, beta-unsaturated carbonyl compounds such as trans-4-phenyl-3-buten-2-one and trans-4-hydroxynon-2-enal. The latter is a cytotoxic product resulting from lipid peroxidation of polyunsaturated fatty acids, and the cardiac isozyme may play a physiologically significant role with glutathione conjugation of this compound. In addition to the near-neutral isozyme, acidic forms with isoelectric points of 4.9, 5.2 and 5.5 were partially purified; some of them are considered to consist of subunits immunologically related to transferase X.

Animals↗
PubMed1986

Function dependent changes in the subcellular distribution of high energy phosphates in fast and slow rat skeletal muscles.

Function dependent changes in the subcellular distribution of ATP, ADP, creatine phosphate (CrP) and creatine (Cr) in rat fast-twitch gastrocnemius and slow-twitch soleus muscles were studied by fractionation of freeze-clamped and freeze-dried tissue in non-aqueous solvents. During 5 min of isotonic contraction of gastrocnemius muscles the mitochondrial content of total creatine [sigma(CrP + Cr)] decreases by 9.5 nmol/mg total protein whereas there is an increase in extramitochondrial total creatine by 12.3 nmol/mg total protein, indicating a net transfer of approximately 10 nmol total creatine/mg total protein/5 min across the mitochondrial inner membrane. During short-term stimulation (6 s) of gastrocnemius muscles the socalled "additionally-bound ADP" correlates not only with force (Hebisch et al. 1984) but also with filament overlap. This confirms the previous suggestion that "additionally-bound ADP" represents actomyosin-ADP-complexes. Following long-term stimulation (10 s), the rate of decay of force is at least two orders of magnitude faster than that of "additionally bound ADP". This indicates a decrease of actomyosin-ADP complexes due to formation of myosin-ADP complexes. Short-term stimulation (6 s) of slow-twitch soleus muscles does not lead to any force-dependent change in the content of "additionally-bound ADP", similar to the finding in long-term contracting gastrocnemius muscles. Denervation of soleus muscles leads to a decrease in "additionally-bound ADP" to values comparable to those found in resting fast-twitch gastrocnemius muscles.

Adenosine Diphosphate↗
PubMed1986

Mechanism and relevance of glutathione mutagenicity.

The ubiquitous tripeptide glutathione (GSH) has previously been shown to be mutagenic to Salmonella typhimurium TA100 when incubated with kidney subcellular fractions at physiological concentrations (Glatt et al., 1983). Here we report that the mutagenic effect of GSH can be inhibited by the use of the gamma-glutamyl-transpeptidase (gamma-GT) inhibitor anthglutin and by the metal chelators bathocuproine, EDTA and diethyldithiocarbamate. As the chelating agents did not inhibit gamma-GT activity this suggested that the mechanism underlying the mutagenic effect of GSH was at least a two-step process, dependent upon the cleavage of GSH by gamma-GT and the presence of either free transition metals or those contained in enzymes such as glutathione oxidase. As gamma-GT is located on the outer surface of kidney tubule cells and is therefore exposed to relatively low concentrations of GSH, and the precise physiological control of levels of transition metals, this mechanism is unlikely to occur in vivo.

Animals↗
PubMed1985

Oxidative stress: damage to intact cells and organs.

Oxidative cell damage can be monitored by detection of (a) photoemission of singlet molecular oxygen formed from radical interactions (so-called low-chemical chemiluminescence), (b) end products of lipid peroxidation, such as ethane, and (c) glutathione disulphide release. These methods, preferably used in a complementary fashion, provide insight into the pro-oxidant-antioxidant balance in the intact cell or organ. Recent work from this laboratory on the metabolism of hydroperoxides and aldehydes as well as on redox cycling of the quinone menadione is presented. The comparison of GSSG transport systems in liver and heart reveals a limitation of capacity in the latter, thus making GSSG export potentially critical in the heart. As part of an inter-organ feedback system between extrahepatic tissues and liver, the newly described hormone stimulation of GSH release from liver is also presented.

Aldehydes↗
PubMed1985

Role of plasma membrane transport in hepatic glutamine metabolism.

In livers of fed rats and in perfused livers supplied with a physiological portal glutamine concentration of 0.6 mM, the mitochondrial and cytosolic glutamine concentrations are 20 mM and 7 mM, respectively, thus, the mitochondrial/cytosolic glutamine concentration gradient is 2-3. Uptake and release of glutamine by periportal and perivenous hepatocytes occurs predominantly by an Na+-dependent transport system (so-called system 'N'). Histidine in near-physiological concentrations inhibits both glutamine uptake by periportal hepatocytes and its release by perivenous hepatocytes. This is not due to an inhibition of glutamine-metabolizing enzymes by histidine or its metabolites. With physiological portal glutamine concentrations (0.6 mM), stimulation of glutaminase flux or of glutamine transaminase flux is followed by a decrease of hepatic glutamine levels to about 80% or 30%, respectively, glutamine levels are further decreased to 50% or 20% in the presence of histidine. When glutamine is synthesized endogenously (no glutamine added), the histidine-induced inhibition of glutamine release is paralleled by a 210% increase of the hepatic tissue level of glutamine. In experiments with and without methionine sulfoximine and in the absence of added glutamine, the glutamine content in the small perivenous hepatocyte population containing glutamine synthetase is estimated to be about 3.5 mumol/g wet weight and that in the periportal hepatocytes as low as 0.1 mumol/g wet weight. In contrast to the prevailing view, it is concluded that glutamine transport across the plasma membrane of hepatocytes is a potential regulatory site in glutamine degradation and synthesis, especially under the influence of effectors like histidine.

Amino Acids↗
PubMed1985

A novel biologically active selenoorganic compound--IV. Protective glutathione-dependent effect of PZ 51 (ebselen) against ADP-Fe induced lipid peroxidation in isolated hepatocytes.

PZ 51 (Ebselen) is capable of inhibiting ADP-Fe-induced lipid peroxidation in isolated hepatocytes, assessed by generation of low-level chemiluminescence, and the formation of alkanes (ethane, n-pentane) and malondialdehyde. The sulphur analog PZ 25 is much less active. PZ 51 is ineffective in glutathione-depleted hepatocytes that were obtained after phorone pretreatment of the animals. However, other antioxidants, like (+)-cyanidanol-3 or diethyldithiocarbamate are effective in inhibiting lipid peroxidation also in glutathione-depleted cells. The results are attributed to a GSH peroxidase-like activity of PZ 51. Addition of the chemically synthetized compound, PZ 51-SG, a glutathione adduct of PZ 51, does not provide extra protection in GSH-depleted cells in comparison to the parent compound, PZ 51. Dithioerythritol exhibits a protective effect, and PZ 51 is more efficient than in the absence of this thiol reductant. Dithioerythritol also exerts an effect in glutathione-depleted cells. A peroxidase-like activity using DTE as reductant is demonstrated in vitro.

Adenosine Diphosphate↗
PubMed1985

Hepatic thiol and glutathione efflux under the influence of vasopressin, phenylephrine and adrenaline.

Thiol and glutathione (GSH) efflux across the sinusoidal plasma membrane in isolated perfused rat liver was stimulated by addition of hormones such as vasopressin, phenylephrine and adrenaline, whereas glucagon or dibutyryl cyclic AMP were without effect. Phenylephrine and adrenaline effects were sensitive to prazosin and phentolamine, respectively. The increase in thiol efflux was largely accounted for by an increase in GSH efflux. Thiol efflux and the hormone effects were abolished in GSH-depleted liver. Biliary GSH efflux was diminished upon hormone addition. The newly discovered hormone-dependence of GSH release across the sinusoidal plasma membrane may explain the known loss of GSH during conditions of experimental shock (traumatic or endotoxin) and stress and peripheral inflammation.

Animals↗
PubMed1985

Oxygen dependence and subcellular partitioning of hepatic menadione-mediated oxygen uptake. Studies with isolated hepatocytes, mitochondria, and microsomes from rat liver in an oxystat system.

Using an oxystat system, menadione (2-methyl-1,4-naphthoquinone)-mediated oxygen uptake was investigated in isolated rat hepatocytes, in malate/glutamate-supplemented mitochondria, and in NADPH-reduced microsomes at steady-state oxygen partial pressures (pO2) between 0.1 to 100 mm Hg (0.2-150 microM O2). Menadione-mediated stimulation of oxygen uptake was half-maximal at pO2 of 0.5, 0.2, and 0.9 mm Hg, respectively. In hepatocytes and mitochondria half-maximal concentrations of menadione were 15 and 4 microM. However, in microsomes saturation with menadione was not reached at concentrations up to 300 microM. Antimycin A inhibited menadione-mediated oxygen uptake in hepatocytes and mitochondria by about three-fourths, while rotenone was without inhibitory effect; KCN inhibited practically completely. In mitochondria menadione-stimulated oxygen uptake was significantly inhibited by dicoumarol but further enhanced by the addition of ADP, even in the presence of rotenone. The results suggest that menadione-mediated hepatocellular oxygen uptake proceeds almost independently of pO2 in most regions of the liver lobule but that in areas of low pO2 such as the centrolobular regions limitation by oxygen may occur. They also demonstrate that in the intact hepatocyte menadione-mediated oxygen uptake predominantly (greater than 90%) results from electron transfer in the mitochondrial respiratory chain by menadione.

Animals↗
PubMed1985

Effect of age and ambient temperature on n-pentane production in adult housefly, Musca domestica.

The objective of this study was to examine the relationship between lipid peroxidation and aging in the male housefly. Metabolic rate of flies is known to be higher and life span shorter at elevated ambient temperature. Evolution of n-pentane and level of thiobarbituric acid (TBA) reactive material were used as indicators of lipid peroxidation. n-Pentane accumulated by houseflies in vivo and by whole body homogenates of houseflies, in response to tert-butyl hydroperoxide (1 mM), increased with age. n-Pentane accumulation in vivo was markedly higher at higher ambient temperature. Furthermore, n-pentane generated by flies in vivo and by fly homogenates in vitro tended to be lower in flies raised at a lower ambient temperature. TBA-reactive material, elicited by tert-butyl hydroperoxide, was augmented in older flies, but no significant difference was found between flies aged at different ambient temperatures. Analysis of fatty acids in housefly homogenates indicated an age-associated increase in the ratio of polyunsaturated to saturated fatty acids.

Aging↗
PubMed1985

Oxidative stress: excited oxygen species and enzyme activity.

The metabolic role of aldehydes, hydroperoxides, and quinones was investigated with emphasis on oxidative transitions involving oxygen free radicals and associated with enzymatic activities. The oxidative metabolism of aldehydes (originating either from ethanol oxidation, or monoamine oxidase activity, or oxidative breakdown of lipid hydroperoxides during lipid peroxidation) is a source of alkane production and low-level chemiluminescence. Since both parameters reflect cellular oxidative conditions, it can be inferred that side-products of aldehyde oxidase activity might participate in the link between the initial enzymatic oxidation of aldehyde and the occurrence of oxidizing species leading to chemiluminescence and alkane production. The metabolism of hydroperoxides was considered under two different aspects: first, the hydroperoxide reduction, within the frame of a detoxication mechanism, as mediated by a selenoorganic compound PZ-51 that displays glutathione peroxidase-like activity and an antioxidant activity; second, the enzyme-catalyzed disproportionation of hydroperoxides as a source of a potent oxidizing equivalent, singlet molecular oxygen. The cytotoxicity of quinones, utilized in therapeutic agents such as anticancer drugs, is believed to be related to oxidative stress due to the formation of the superoxide radical and subsequent more reactive oxygen species. The enzyme-catalyzed one-electron reduction of menadione seems to play a substantial role in the development of cytotoxic effects, at variance with the 2-electron reduction of the quinone. The observation of low-level chemiluminescence under conditions which favor the one-electron reduction process or which diminished the two-electron reduction process indicates the practicability of low-level chemiluminescence measurements in monitoring changes in quinone metabolism and related cytotoxic effects.

Aldehydes↗