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Biomedical subjects

H Stam

Publications and source records attributed to H Stam.

At least 91 records · Page 5Linked to original sources
PubMed1981

Comparative study of chylomicron and fatty acid utilization in small intestine and heart.

Chylomicrons were isolated from the urine of rats after a surgical procedure in which the cysterna chyli was connected with the right ureter. The fatty acids of the chylomicrons served as a respiratory substrate for rat heart and not for rat small intestine during in vitro vascular perfusions. The reason for the absence of chylomicron utilization in small intestine was found to be the virtual absence of lipoprotein lipase from this organ. Both heart and small intestine oxidized oleate complexed to albumin. Increasing the molar ratio of fatty acid to albumin from 3 to 6 did not affect the rate of fatty acid oxidation in heart, but increased fatty acid oxidation in small intestine.

Albumins↗
PubMed1981

Control of lipolysis in triglyceride-enriched rat hearts.

Hormonal and metabolic regulation of endogenous triglyceride hydrolysis was studied in triglyceride-enriched hearts obtained from rats fed 3 days with a trierucate-rich diet. Endogenous lipolysis was determined by measuring glycerol release during in vitro perfusion of the hearts. It appeared that there was a direct relation between the contractile state of the heart, the rate of glycerol release in the coronary effluent and the Ca2+ concentration in the perfusion medium. During Ca2+-free perfusion, 2,4-dinitrophenol stimulated oleate oxidation and this, as well as the addition of 2 x 10(-7) M glucagon, induced a marked stimulation of lipolysis. Insulin did not affect glucagon- and norepinphrine-stimulated lipolysis during substrate-free perfusion. The presented experiments point out that in lipid-enriched rat hearts the activity of the tissue lipase may be controlled by the rate of beta-oxidation and re-esterification of the liberated fatty acid, as well as by a shift to utilization of carbohydrate instead of fatty acids for energy supply.

2,4-Dinitrophenol↗
PubMed1976

Isolation and partial characterization of chick brain synaptic plasma membranes.

An isolation procedure for synaptic plasma membranes from whole chick brain is reported that uses the combined flotation-sedimentation density gradient centrifugation procedure described by Jones and Matus (Jones, D. H. and Matus, A. I. (1974) Biochim. Biophys. Acta 356, 276-287) for rat brain. The particulate of the osmotically shocked and sonicated crude mitochondrial fraction was used for a flotation-sedimentation gradient step. Four fractions were recovered from the gradient after 30 min centrifugation. The fractions were identified and characterized by electron microscopy and by several markers for plasma membrane and other subcellular organelles. Fraction 2 was recovered from the 28.5-34% (w/v) sucrose interphase and contained the major part of the activities of the neuronal plasma membrane marker enzymes. The specific activities of the (Na+ +K+)-activated ATPase (EC 3.6.1.3), acetylcholinesterase (EC 3.1.1.7) and 5'-nucleotidase (EC 3.1.3.5) were, respectively, 4.5, 2.0 and 1.2 times higher than in the homogenate. However, Fraction 2 also contained considerable amounts of activities of putative lysosomal and microsomal markers in addition to lower amounts of mitochondrial and myelin markers. Although no prepurification of synaptosomes from the crude mitochondrial fraction was performed, the synaptic plasma membranes obtained showed many properties analogous to similar preparations from rat brain described in recent years.

Acetylcholinesterase↗
PubMedDate not supplied

Hormones and triacylglycerol metabolism under normoxic and ischemic conditions.

Fatty acids, the preferred substrate in normoxic myocardium, are derived from either exogenous or endogenous triacylglycerols. The supply of exogenous fatty acids is dependent of the rate of lipolysis in adipose tissue and of the lipoprotein lipase activity at the coronary vascular endothelium. A large part of the liberated fatty acids is reesterified with glycerol-3-phosphate and converted to triacylglycerols. Endogenous lipolysis and lipogenesis are intracellular compartmentalized multienzyme processes of which individual hormone-sensitive steps have been demonstrated in adipose tissue. The triacylglycerol lipase is the rate-limiting enzyme of lipolysis and glycerol-3-phosphate acyltransferase and possibly phosphatidate phosphohydrolase are the rate-limiting enzymes of lipogenesis. The hormonal regulation of both processes in heart is still a matter of dispute. Triacylglycerol lipase activity in myocardial tissue has two intracellular sources: 1. the endoplasmic reticular and soluble neutral lipase, and 2. the lysosomal acid lipase. Studies in our laboratory have indicated that whereas lipolysis is enhanced during global ischemia and anoxia, overall lipolytic enzyme activities in heart homogenates were not altered. In addition we were unable to demonstrate alterations in tissue triacylglycerol content and glycerol-3-phosphate acyltransferase activity under these conditions. Lipolysis, is subject to feedback inhibition by product fatty acids. Therefore all processes leading to an increased removal of fatty acids from the catalytic site of the lipase will stimulate lipolysis. These studies will be reviewed. In addition, studies from our department have demonstrated the capacity of myocardial lysosomes to take up and degrade added triacylglycerol-particles in vitro. Such a process, stimulated by Ca2+ and stimulated by acidosis, offers another physiological target for hormone actions.

Animals↗
PubMedDate not supplied

Involvement of lysosome-like particles in the metabolism of endogenous myocardial triglycerides during ischemia/reperfusion. Uptake and degradation of triglycerides by lysosomes isolated from rat heart.

The hormonal regulation and enzymatic basis of endogenous lipolysis in heart are not yet completely elucidated. The lysosomal fraction from rat heart appeared to be markedly enriched in triglycerides and a significant reduction in triglycerides in this fraction was found after prolonged perfusion or stimulation of lipolysis with glucagon. The enhanced rate of lipolysis, measured as glycerol release from the isolated perfused rat heart, was abolished 10-15 min after continuous glucagon administration. Omission of glucagon for another 60 min restored the ability of glucagon to stimulate lipolysis, indicating the limited availability of endogenous triglycerides and the presence of a transfer-system for triglycerides from a non-metabolically active pool to a metabolically active pool. The enhanced lipolysis induced by low-flow ischemia was found to be inhibited by the lysosomotropic agent methylamine (5 mM). Methylamine-perfusion during low-flow ischemia was accompanied by an increased recovery of myocardial triglycerides in the lysosomal fraction. The possible role of lysosome-like particles in myocardial triglyceride homeostasis was further investigated by studying the kinetics of uptake and degradation of labeled triglycerides by membrane-particles recovered in the subcellular fraction enriched with lysosomal marker enzymes. It appeared that isolated lysosomal membranes take up added triglycerides at an average rate of 30 nmoles/min/g protein. The bulk of these triglycerides taken up is stored whereas 20% is degraded to diglycerides and free fatty acids. More than 90% of the free fatty acids formed were released from the lysosomes into the supernatant. The uptake and degradation of triglyceride-filled liposomes by isolated myocardial lysosomes was inhibited during incubation with methylamine (5 mM). On the other hand, a lowering of pH during in vitro incubation increased the rate of uptake and degradation of added triglycerides by isolated lysosomes. These results indicate that lysosomes or lysosome-like particles are involved in the enhanced lipolysis during myocardial ischemia.

Acid-Base Equilibrium↗
PubMedDate not supplied

Enhanced lipolysis of myocardial triglycerides during low-flow ischemia and anoxia in the isolated rat heart.

We studied lipolysis in the isolated rat heart, measured as glycerol release during anoxia, low-flow ischemia and subsequent reperfusion. It was found that the rate of lipolysis was enhanced during ischemia/anoxia while the lipase activities in tissue extracts involved in the myocardial lipolysis and the amount of triglycerides were not affected. This indicates the dominant occurrence of a lipolysis-reesterification principle in ischemic and anoxic tissue. A common observation of ischemia/anoxia is an increase in the tissue NADH/NAD+ ratio. Therefore we investigated the effect of lactate and malate, both of which enhance the tissue redox state on myocardial lipolysis. Perfusion in the presence of lactate (10 mM) and malate (10 mM) both stimulated myocardial lipolysis by about five times. This suggests that the rate of reesterification of product fatty acids to triglycerides, which is determined by the NADH/NAD+ ratio, because of the increased formation of glycerol 3-phosphate from dihydroxy acetone phosphate, plays an important role in the regulation of lipolysis. The existence of triglyceride-fatty acid-triglyceride cycle is discussed.

Animals↗
PubMedDate not supplied

Localization and function of myocardial lipolysis.

Mobilization of triacylglycerol stored in heart cells is accomplished by the combined action of lysosomal (acid) lipase and microsomal monoacylglycerol lipase or carboxylesterase. Non(heparin)-releasable neutral or alkaline lipase is similar to non(readily)-releasable lipoprotein lipase (LPL). The enzyme is mainly localized extracellularly. Non(readily)-releasable LPL probably represents LPL in caveola or vacuolae of vascular endothelium and/or LPL on myocardial interstitium. It contributes to the uptake of lipoprotein constituents in heart cells. Glycerol, an endproduct of lipolysis, is not a reliable marker for the net mobilization of lipid stored in heart cells. It is formed both intra- and extracellularly, and does not reflect the rate of oxidation of part of free fatty acids formed.

Animals↗