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R G Slee

Publications and source records attributed to R G Slee.

At least 19 recordsLinked to original sources
PubMed1986

Possible involvement of the lipid-peroxidation product 4-hydroxynonenal in the formation of fluorescent chromolipids.

The effects of the lipid-peroxidation product 4-hydroxynonenal on the formation of fluorescent chromolipids from microsomes, mitochondria and phospholipids were studied. Incubation of freshly prepared rat liver microsomes or mitochondria with 4-hydroxynonenal results in a slow formation of a fluorophore with an excitation maximum at 360 nm and an emission maximum at 430 nm. The rate and extent of the development of the 430 nm fluorescence can be significantly enhanced by ADP-iron (Fe3+). With microsomes, yet not with mitochondria. NADPH has a catalytic effect similar to that of ADP-iron. Fluorescent chromolipids with maximum excitation and emission at 360/430 nm are also formed during the NADPH-linked ADP-iron-stimulated lipid peroxidation. Phosphatidylethanolamine and phosphatidylserine react with 4-hydroxynonenal revealing a fluorophore with the same spectral characteristics as that obtained in the microsomal and mitochondrial system. The findings suggest that the fluorescent chromolipids formed by lipid peroxidation are not derived from malonaldehyde, but are formed from 4-hydroxynonenal or similar reactive aldehydes via a NADPH and/or ADP-iron-catalysed reaction with phosphatidylethanolamine and phosphatidylserine contained in the membrane.

Adenosine Diphosphate↗
PubMed1986

Ferritin, a physiological iron donor for microsomal lipid peroxidation.

In the process of lipid peroxidation of microsomes induced either by oxygen radicals generated by xanthine oxidase or by NADPH, ferritin is able to donate the necessary iron. The amount of ferritin necessary to catalyze the process of lipid peroxidation is in the physiological range. In contrast to the finding with phospholipid liposomes, catalase hardly stimulates the lipid peroxidation of microsomes.

Animals↗
PubMed1986

Comparison of the inactivation of microsomal glucose-6-phosphatase by in situ lipid peroxidation-derived 4-hydroxynonenal and exogenous 4-hydroxynonenal.

1) The effect of 4-hydroxynonenal and lipid peroxidation on the activities of glucose-6-phosphatase and palmitoyl CoA hydrolase were studied. 2) 4-Hydroxynonenal inactivates glucose-6-phosphatase but has no effect on palmitoyl-CoA hydrolase. These effects are similar with those observed during lipid peroxidation of microsomes. 3) The inhibition of glucose-6-phosphatase by 4-hydroxynonenal can be prevented by glutathione but not by vitamin E. The inactivation of glucose-6-phosphatase during lipid peroxidation is prevented by glutathione and delayed by vitamin E. 4) The formation of 4-hydroxynonenal during lipid peroxidation was followed in relation to the inactivation of glucose-6-phosphatase. At 50% inactivation of glucose-6-phosphatase the 4-hydroxynonenal concentration was 1.5 microM. To obtain 50% inactivation of glucose-6-phosphatase by added 4-hydroxynonenal a concentration of 150 microM or 300 microM was needed with a preincubation time of 30 and 60 min, respectively. 5) It is concluded that the glucose-6-phosphatase inactivation during lipid peroxidation can be due to the formation of 4-hydroxynonenal. The formed 4-hydroxynonenal which inactivates glucose-6-phosphatase is located in the membrane. If this mechanism is valid it implies that a functional SH group of glucose-6-phosphatase is layered in the membrane. However, an inactivation of glucose-6-phosphatase by desintegration of the membrane by lipid peroxidation cannot be ruled out.

Aldehydes↗
PubMed1985

The involvement of iron and lipid peroxidation in the pathogenesis of HCB induced porphyria.

Hexachlorobenzene (HCB) induces a porphyria characterized by a diminished activity of the enzyme uroporphyrinogen decarboxylase (URO-D), presumably due to inactivation by reactive metabolites of HCB. We studied the effect of iron on HCB porphyria in female rats, to determine whether the iron dependent process of lipid peroxidation was involved in the pathogenesis of porphyria. We showed that malondialdehyde formation is increased in rat liver tissue of porphyric rats and that high molecular weight proteins due to cross-linking are formed. We also showed that the induction of porphyria by HCB is dependent on the presence of iron. Our findings suggest that lipid peroxidation is involved in the toxicity of HCB and that the aggravating effects of iron on HCB are mediated by lipid peroxidation.

Animals↗
PubMed1985

Studies on cumene hydroperoxide-induced lipid peroxidation in the isolated perfused rat heart.

In the isolated, perfused rat heart, lipid peroxidation, induced by cumene hydroperoxide (Cum OOH), is accompanied by the release of malondialdehyde (MDA). Using a modified perfusion technique resulting in the separate collection of coronary and interstitial effluent, it can be shown that upon Cum OOH (0.5 mM) perfusion there is an immediate release of MDA in the coronary effluent and a delayed release in the interstitial fluid, indicating the susceptibility and coronary vascular tissue towards free radical-induced lipid peroxidation. Perfusion with Cum OOH leads to an initial increase of the coronary flow and a depressed contractility followed by a cardiac arrest concomitantly with the onset of MDA release in the interstitial fluid. Finally, during prolonged perfusion the coronary flow diminishes and contracture of the heart muscle ('stone heart') develops. These phenomena resemble those occurring during the 'calcium paradox'. Although the contractility diminishes immediately after the perfusion with Cum OOH the tissue ATP level and energy charge (formula; see text) remain constant. From the moment of cardiac arrest the ATP and creatine phosphate levels gradually decrease and the energy charge drops simultaneously with the appearance of MDA in the interstitial fluid. In contrast to the calcium paradox there is no simultaneous increase in the myocardial AMP level. Various mitochondrial enzymes (cytochrome c oxidase, monoamine oxidase, carnitinepalmitoyltransferase I and palmitoyl CoA synthetase) were tested and not affected by Cum OOH perfusion. During the development of contracture after 20 min of Cum OOH perfusion massive contraction band necrosis of cardiac tissue occurs. However, overall protein release is lower when compared with the protein release during the calcium paradox.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenine Nucleotides↗
PubMed1984

Biochemical genetics of the Lapland dog model of glycogen storage disease type II (acid alpha-glucosidase deficiency).

A recently described canine model (Lapland dog) of glycogen storage disease type II (GSD II, Pompe disease, acid alpha-glucosidase deficiency) was identified with several biochemical genetic methods. Complementation studies in which fibroblasts from a GSD II dog were fused with fibroblasts derived from control dogs and from human patients with different clinical forms of the disease did not lead to restoration of acid alpha-glucosidase activity in the heterokaryon cell populations. These results indicate that acid alpha-glucosidase deficiency is the primary defect in canine GSD II and that there is a close genetic parallelism with human GSD II. Immunotitration analysis of the residual acid alpha-glucosidase activity in the canine GSD II fibroblasts and liver demonstrated that this residual activity was not due to acid alpha-glucosidase enzyme, in which respect canine GSD II was similar to the infantile form of the human disease. Double immunodiffusion studies showed the presence of catalytically inactive acid alpha-glucosidase enzyme protein in canine GSD II. This is consistent with a structural gene mutation. It is concluded that canine GSD II in the Lapland dog is a homologous model of the infantile form of human GSD II, a conclusion in concordance with clinical and pathological studies.

Animals↗
PubMed1983

The effect of (13 OOH) linoleic acid on human erythrocytes and on erythrocyte ghosts.

Peroxidized linoleic acid (13 OOH) induces lipid peroxidation, measured as malondialdehyde in erythrocyte ghosts and intact erythrocytes. This process can be inhibited by desferal, thiourea and butylated hydroxytoluene. During the lipid peroxidation process fluorescent chromolipids are formed. The synthesis of these chromolipids can be blocked by desferal. Peroxidized linoleic acid gives cross-linking of the proteins present in the membranes leading to the formation of higher molecular weight proteins. Also in the lower molecular weight region of SDS-electrophoresis a protein band is formed. The cross-linking process of the membrane protein is most effectively blocked by thiourea, to some extent by butylated hydroxytoluene and hardly at all by desferal. Addition of peroxidized linoleic acid to intact erythrocytes leads to a drop in the glutathione level (analogous to cumene hydroperoxide). In the presence of glucose the glutathione level can be restored.

Erythrocyte Membrane↗
PubMed1983

Lipid peroxidation of human erythrocyte ghosts induced by organic hydroperoxides.

Isolated human erythrocyte ghosts perform lipid peroxidation, measured as malondialdehyde, induced by cumene hydroperoxide and t-butyl hydroperoxide but not by H2O2. In contrast to Ames et al. (Ames, B.N., Cathcart, R., Schwiers, E. and Hochstein, P. (1981) Proc. Natl. Acad. Sci. 78, 6858-6862), no inhibition is found by uric acid, only an increase in lag-time of the malondialdehyde production. In parallel with the malondialdehyde production, fluorescent chromolipids are also formed. Both processes are blocked by the addition of desferal, a potent iron chelator. The malondialdehyde production is also inhibited by the OH radical scavenger, thiourea, and by the anti-oxidant, butylated hydroxytoluene. Treatment of erythrocyte ghosts with cumene hydroperoxide or t-butyl hydroperoxide leads to the genesis of high-molecular-weight protein, but not with H2O2. The appearance of high-molecular-weight proteins is accompanied by disappearance of protein bands, e.g., the alpha- and beta-spectrin band, the anion-exchanger and some other smaller bands. Furthermore, a protein band is formed in the lower-molecular-weight region. 4. The addition of desferal does not reveal any blockade of the high-molecular-weight protein genesis. In contrast, a marked diminution of high-molecular-weight proteins is observed by the addition of thiourea, accompanied by a protection of the protein bands which would otherwise disappear. Similar results are obtained with butylated hydroxytoluene. 5. It is concluded that under oxidative stress the process of high-molecular-weight protein genesis can occur independently of the lipid peroxidation process, measured as the revealing of malondialdehyde.

Blood Proteins↗
PubMed1982

Canine glycogen storage disease type II. A biochemical study of an acid alpha-glucosidase-deficient Lapland dog.

A biochemical study was performed in a Lapland dog suspected of glycogen storage disease type II (acid alpha-glucosidase deficiency, Pompe's disease). Glycogen content was substantially elevated in heart and skeletal muscle but not in the liver. Severely reduced activities of acid alpha-glucosidase (EC 3.2.1.20) were found in heart, skeletal muscle, liver and cultured tongue fibroblasts. The deficiency was located in the glycoprotein fraction, which supported its lysosomal origin. The electrophorogram showed after acid incubation that the affected dog was missing the activity band, while after neutral incubation the pattern was similar to control. The obtained biochemical data are compared with the known data of the human pathology.

Animals↗
PubMed1982

On the lipid peroxidation of rat liver hepatocytes, the formation of fluorescent chromolipids and high molecular weight protein.

1. The formation of malondialdehyde by intact hepatocytes, induced by ADP/Fe3+ or cumene hydroperoxide, can be inhibited by the addition of thiourea. This may indicate that hydroxyl radicals are involved in this process. 2. Lipid peroxidation of intact hepatocytes leads to the formation of fluorescent chromolipids. When similar amounts of malondialdehyde are formed by either ADP/Fe3+ or cumene hydroperoxide, the lipid peroxidation induced by cumene hydroperoxide generates more fluorescent chromolipids than does the lipid peroxidation induced by ADP/Fe3+. 3. The formation of chromolipids is accompanied by the genesis of high molecular weight protein. With cumene hydroperoxide more high molecular weight protein is formed than with ADP/Fe3+. 4. It can be concluded that the defense system against lipid peroxidation of intact hepatocytes does not prevent the formation of lipofuscin-like chromolipids and high molecular weight protein as found earlier in microsomes. Cumene hydroperoxide, at least in this system, can be considered as an effective inducer of chromolipids.

Adenosine Diphosphate↗
PubMed1981

Identification of heterozygotes for glycogenosis 2 (acid maltase deficiency).

In 21 obligate and 9 possible heterozygotes for acid maltase deficiency (AMD) (glycogenosis 2, Pompe's disease), different methods of identifying heterozygotes have been studied. Heterozygosity could not be demonstrated by physical examination, serum CPK assays, morphological examination of muscle biopsy (including light-microscopy, histochemistry and electron-microscopy), or by ultrastructural examination of a skin biopsy. Heterozygotes could be identified to a large, but still limited extent, by measuring the acid alpha-glucosidase activity in urine, cultivated fibroblasts, leucocytes, or skeletal muscle. Heterozygotes for the generalized from of AMD could not be distinguished from those for the muscular form. The limitations of heterozygote identification by means of enzyme assays are discussed, and some practical aspects for genetic counselling are mentioned.

Female↗
PubMed1981

A family with different clinical forms of acid maltase deficiency (glycogenosis type II): biochemical and genetic studies.

In the same family, the generalized or infantile form of acid maltase deficiency (glycogenosis type II, Pompe disease) and the muscular or adult-onset form affected different individuals. Autosomal-recessive inheritance for the two clinical forms was demonstrated in this family by assay of acid alpha-glucosidase in muscle, lymphocytes, cultured fibroblasts, and urine of asymptomatic relatives. Current biochemical techniques do not discriminate between persons heterozygous for the generalized form and those heterozygous for the muscular form. To explain the coexistence of both forms in the same family, the infant with the generalized form or her grandfather with the muscular form must have been a genetic compound of different mutant alleles for acid alpha-glucosidase.

Adult↗
PubMed1980

Lipid peroxidation of rat liver microsomes.

1. The NADPH-dependent lipid peroxidation process was studied with microsomes and also the effects of addition of superoxide dismutase, catalase and thiourea. Only catalase and thiourea were able to inhibit lipid peroxidation. It seems that the initiating radical is the OH. radical formed by the Fenton reaction. 2. During lipid peroxidation glucose-6-phosphatase is inactivated, whilst the microsomal enzyme palmitoyl-CoA hydrolase is practically not affected. Because glucose-6-phosphatase activity decreases during ageing and palmitoyl-CoA hydrolase does not, a possible relationship with the ageing process is thought to exist. 3. Chromolipids are formed by the NADPH-dependent lipid peroxidation. These chromolipids have the same excitation-emission spectra as described for lipofuscin. The formation of these chromolipids is blocked by the addition of catalase and thiourea. 4. High-molecular weight proteins are formed during the NADPH-dependent lipid peroxidation. This process can be associated with the inactivation of enzymes. Also polymerisation is prevented by catalase and thiourea.

Animals↗
PubMed1980

Isoenzymes of human phosphofructokinase.

Human liver phosphofructokinase has been isolated in order to obtain antibodies against human liver phosphofructokinase. With the antiserum it could be shown in human liver that two types of phosphofructokinase exist with no subunits in common. Leucocytes, erythrocytes and kidney also possess the liver (L-) type subunit and a small amount of L-type is present in brain. There is no L-type of phosphofructokinase present in muscle and heart tissue.

Brain↗
PubMed1979

McArdle's disease: a study on the molecular basis of two different etiologies of myophosphorylase deficiency.

Three patients with myophosphorylase deficiency were investigated. Two had no detectable activity, while one had 1% residual activity. The patient with 1% residual activity had 40% of the normal amount of myophosphorylase protein. No myophosphorylase protein could be detected in the other two cases. A precipitin band in the Ouchterlony double immunodiffusion test was not present in any case. This study showed that modifying the normal enzyme (without changing the molecular weight) changed the immunoprecipitin activity of the phosphorylase protein. Therefore, immunoprecipitation is not a valid technique for differentiation of the variants of myophosphorylase deficiency, and another method, for example SDS-electrophoresis, should be applied.

Electrophoresis, Polyacrylamide Gel↗