VLDL substrate properties and efficiency of their metabolic transformation by LPL

A study was made of the regulation of the triglyceride hydrolysis catalysed by LPL from bovine milk, by the apoproteins from human plasma VLDL. Both isolated apolipoproteins, and those found on the surface of plasma VLDL particles, were investigated. A concentration-dependent activating action of apo C-II on the hydrolysis of emulsified triolein, and uncompetitive inhibition of VLDL triglyceride hydrolysis by apo C-III were found. It is suggested that VLDL lipolysis might be controlled in vivo through the variation of the relative surface content of these enzymatic activity modulators.     

Studies on the late-pre-beta-lipoprotein of human serum.

Very low density lipoprotein (VLDL) isolated from 3 healthy normolipidaemic subjects had a raised VLDL cholesterol to triglyceride ratio. The VLDL fractions gave 2 pre-beta-bands on agarose gel electrophoresis. Family study of the subjects appears to indicate sex linkage of this trait and a possible polygenic type of inheritance.     

Familial hypobetalipoproteinemia: genetics and metabolism

Familial hypobetalipoproteinemia (FHBL), an autosomal dominant disorder, is defined as <5^th percentile LDL-cholesterol or apolipoprotein (apo) B in the plasma. FHBL subjects are generally heterozygous and asymptomatic. Three genetic forms exist: (i) premature stop codon specifying mutations of APOB; (ii) FHBL linked to a susceptibility locus on the chromosome 3p21; and (iii) FHBL linked neither to APOB nor to the chromosome 3p21. In heterozygous apoB-defective FHBL, the hepatic VLDL export system is defective because apoB 100, the product of the normal allele, is produced at ∼25% of normal rate, and truncated apoB is cleared too rapidly. The reduced capacity for hepatic triglyceride export increases hepatic fat three-fold. Indexes of adiposity and insulin action are similar to controls. ‘Knock-in’ mouse models of apoB truncations resemble human FHBL phenotypes. Liver fat in the chromosome 3p21-linked FHBL is normal. Elucidation of the genetic basis of the non-apoB FHBL could uncover attractive targets for lipid-lowering therapy. (See note added in proof.)Received 27 October 2004; received after revision 1 February 2005; accepted 22 February 2005     

Studies on the late-pre-β-lipoprotein of human serum

Summary Very low density lipoprotein (VLDL) isolated from 3 healthy normolipidaemic subjects had a raised VLDL cholesterol to triglyceride ratio. The VLDL fractions gave 2 pre--bands on agarose gel electrophoresis. Family study of the subjects appears to indicate sex linkage of this trait and a possible polygenic type of inheritance.Acknowledgment. This investigation was supported by a grant from University of Nigeria, Nsukka. I am grateful to Prof. Barry Lewis, St. Thomas Hospital Medical School, London, for his advice.     

Marked elevation of HDL-cholesterol in cold-adapted golden hamsters

Golden hamsters with spontaneous hypercholesterolemia at 22 degrees C developed a further increase in plasma cholesterol when they were maintained at 6 degrees C. This hypercholesterolemia was associated with a redistribution of plasma cholesterol between VLDL and HDL. Plasma cholesterol transported in the VLDL decreased while cholesterol in the HDL increased by 45%. The LDL profile was not significantly modified.     

Marked elevation of HDL-cholesterol in cold-adapted golden hamsters

Summary Golden hamsters with spontaneous hypercholesterolemia at 22°C developed a further increase in plasma cholesterol when they were maintained at 6°C. This hypercholesterolemia was associated with a redistribution of plasma cholesterol between VLDL and HDL. Plasma cholesterol transported in the VLDL decreased while cholesterol in the HDL increased by 45%. The LDL profile was not significantly modified.     

Effect of low density lipoprotein and high density lipoprotein on sodium dodecyl sulphate precipitation of very low density lipoprotein from human serum.

In the presence of low density lipoprotein, the sodium sodecyl sulphate(SDS)-very low density lipoprotein(VLDL) complex sedimented, while in the presence of high density lipoprotein the complex floated. This SDS-VLDL aggregate floats at serum triglyceride to cholesterol ratio of 0.7-0.9 and sediments at a ratio of 0.2-0.5.     

Effect of low density lipoprotein and high density lipoprotein on sodium dodecyl sulphate precipitation of very low density lipoprotein from human serum

Summary In the presence of low density lipoprotein, the sodium sodecyl sulphate(SDS)-very low density lipoprotein(VLDL) complex sedimented, while in the presence of high density lipoprotein the complex floated. This SDS-VLDL aggregate floats at serum triglyceride to cholesterol ratio of 0.7–0.9 and sediments at a ratio of 0.2–0.5.     

Hydrolysis of N-phenylacetyl-a-methyl-alpha-amino acids by benzylpenicillinacylase

Enzymatic hydrolysis of several racemic N-phenylacetyl-alpha-methyl-alpha-amino acids containing an additional aliphatic, aromatic or polar substitutent on the chiral carbon atom, has been studied by using benzylpenicillinacylase from Escherichia coli A.T.C.C.9637. Both the rate of hydrolysis and the stereoselectivity were found to be considerably lower than in the case of natural alpha-amino acids. Steric and electronic factors in the side chains influencing the stereoselectivity are discussed. Key words. Benzylpenicillinacylase; enzymatic hydrolysis; alpha-methyl-alpha-amino acids.     

Influence of corpora allata and brain extract on the lipid release from the fat body of termite queen Odontotermes assmuthi

Summary Short term in vitro experiments on the influence of the extracts of corpora allata and brain from the termite queenOdontotermes assmuthi on the lipid release from the fat body into the haemolymph indicated that the extract of corpora allata does not influence the lipid mobilization, whereas the brain extract increases the free fatty acid level in the haemolymph. It is believed that the brain extract stimulates triglyceride hydrolysis in the fat body.     

Cholesteryl ester transfer protein and its inhibition

Cholesteryl ester transfer protein (CETP) is a plasma glycoprotein that facilitates the transfer of cholesteryl esters from the atheroprotective high density lipoprotein (HDL) to the proatherogenic low density lipoprotein cholesterol (LDL) and very low density lipoprotein cholesterol (VLDL) leading to lower levels of HDL but raising the levels of proatherogenic LDL and VLDL. Inhibition of CETP is considered a potential approach to treat dyslipidemia. However, discussions regarding the role of CETP-mediated lipid transfer in the development of atherosclerosis and CETP inhibition as a potential strategy for prevention of atherosclerosis have been controversial. Although many animal studies support the hypothesis that inhibition of CETP activity may be beneficial, negative phase III studies on clinical endpoints with the CETP inhibitor torcetrapib challenged the future perspectives of CETP inhibitors as potential therapeutic agents. The review provides an update on current understanding of the molecular mechanisms involved in CETP activity and its inhibition.     

Diethylmesoxalate hydrate, a new irreversible inhibitor of cholinesterases.

Bovine erythrocyte acetylcholinesterase and human plasma cholinesterase are irreversibly inhibited by diethylmesoxalate hydrate, the inhibition potency being comparable to that of certian insecticidal organophosphates and carbamates. Insect cholinesterases, however, appear to be much less affected by diethylmesoxalate hydrate. The compound was also found to inhibit the hydrolysis of paraoxon by rabbit plasma A-esterase, but in a reversible mode.     

Diethylmesoxalate hydrate,a new irreversible inhibitor of cholinesterases

Summary Bovine erythrocyte acetylcholinesterase and human plasma cholinesterase are irreversibly inhibited by diethylmesoxalate hydrate, the inhibition potency being comparable to that of certain insecticidal organophosphates and carbamates. Insect cholinesterases, however, appear to be much less affected by diethylmesoxalate hydrate. The compound was also found to inhibit the hydrolysis of paraoxon by rabbit plasma A-esterase, but in a reversible mode.     

Relationship of phosphatidylcholine to hydrophobic surfactant on rat intestinal chylomicron secretion

Hydrophobic surfactants such as Poloxalene inhibit triglyceride secretion into lymph by enterocytes. The inhibitory effect of these agents on triglyceride secretion is reversed when lipid presented for absorption is exclusively in the form of phosphatidylcholine (PC) and not triglyceride. The present investigation performed in conscious mesenteric lymph fistula rats was designed to determine whether various mixtures of triglyceride and PC given intraduodenally with Poloxalene would also reverse the inhibitory effect of Poloxalene on triglyceride secretion into lymph. A 50–50 mixture of triolein (TO) and PC resulted in normal triglyceride secretion into lymph. However, when the mixture of lipids was 75-25, TO to PC, results for triglyceride recovery in lymph were considerably reduced. The transport rate for triglyceride into lymph was not as depressed, however, as observed for Poloxalene treated rats given lipid for absorption basically in the triglyceride form. Substitution of phosphatidylethanolamine for PC had no beneficial effect on triglyceride secretion in Poloxalene treated rats. It is concluded that PC can reverse the inhibitory effect of Poloxalene on triglyceride secretion into lymph even when considerable amounts of triglyceride along with PC are presented for absorption.     

3-Hydroxy-3-methylglutaric acid and experimental atherosclerosis in rats

Summary In rats 3-hydroxy-3-methylglutaric acid effectively counteracts the lipemic and atherosclerotic response of massive doses of vitamin D₂. It regressed the formation of atheromatous arterial lesions. Furthermore the significant decrease in serum -lipoprotein levels on HMG treatment could be due to decrease in VLDL triglyceride and cholesterol levels.Acknowledgments. The authors are indebted to Dr.W. Drell, President, Calbiochem, San Diego, California, USA, for generous gift of HMG and Lady Tata Memorial Trust, India, for providing financial assistance to one of us (S.Y.K.Y.).     

Hydrolysis of histones by horse urinary kallikreins

Summary Horse urinary kallikrein was shown to hydrolyze histones from calf thymus and chicken nuclei erythrocytes. The hydrolysis is inhibited by benzamidine and hyposulfite but not by soy-bean trypsin inhibitor; horse plasma kallikrein also produces this hydrolysis.Work supported by grants from Projeto BIOQ/FAPESP (São Paulo), FINEP (Rio) and CNPq (Rio).Fellow from CNPq, from Instituto Biológico (São Paulo).     

Use of 3H-QNB in the isolation of plasma membrane from smooth muscle of the urinary bladder: effect of oxalate on calcium uptake by the membrane fractions

Specific binding of tritiated quinuclidinyl benzilate (3H-QNB) to surface membrane muscarinic receptors was utilized to identify plasma membrane (PM) fractions from smooth muscle of the rabbit urinary bladder. Accumulation of 3H-QNB in the PM fraction was 4-5-fold higher than that in fractions of endoplasmic reticulum (EM) or mitochondria (M). A similar pattern of distribution was found for 5'-nucleotidase. 3H-QNB binding therefore appears to be a suitable marker for plasma membrane of the urinary bladder. Data on ATP-dependent calcium uptake by PM and ER fractions showed that oxalate highly potentiated calcium uptake by both fractions and consequently this feature cannot be used to identify ER fractions specifically.     

Interaction of lipid transfer protein with plasma lipoproteins and cell membranes

Summary The hydrophobic lipid components of lipoproteins, cholesteryl ester and triglyceride, are transferred between all lipoproteins by a specific plasma glycoprotein, termed lipid transfer protein (LTP). LTP facilitates lipid transfer by an exchange process in which cholesteryl ester and triglyceride compete for transfer. Thus, LTP promotes remodeling of the lipoprotein structure, and plays an important role in the intravascular metabolism of these particles and in the lipoprotein-dependent pathways of cholesterol clearance from cells. The properties of LTP, its mechanisms of action, its roles in lipoprotein metabolism, and its modes of regulation are reviewed along with recent data that suggest a possible role for this protein in directly modifying cellular lipid composition.     

Confocal microscopy and biochemical analysis reveal spatial and functional separation between anandamide uptake and hydrolysis in human keratinocytes

The signaling activity of anandamide (AEA) is terminated by its uptake across the cellular membrane and subsequent intracellular hydrolysis by the fatty acid amide hydrolase (FAAH). To date, the existence of an AEA membrane transporter (AMT) independent of FAAH activity remains questionable, although it has been recently corroborated by pharmacological and genetic data. We performed confocal microscopy and biochemical analysis in human HaCaT keratinocytes, in order to study the cellular distribution of AMT and FAAH. We found that FAAH is intracellularly localized as a punctate staining partially overlapping with the endoplasmic reticulum. Consistently, subcellular fractionation and reconstitution of vesicles from membranes of different compartments demonstrated that FAAH activity was localized mainly in microsomal fractions, whereas AMT activity was almost exclusively in plasma membranes. These results provide the first morphological and biochemical evidence to support the view that transport and hydrolysis are two spatially and functionally distinct processes in AEA degradation.Received 11 October 2004; received after revision 24 November 2004; accepted 7 December 2004     

Effect of acetone on VLDL secretion by the isolated rat liver.

High acetone levels occur in uncontrolled diabetes and after isopropanol administration to rats. In both conditions, very low density lipoproteins (VLDL) secretion is depressed. Acetone, hoever, failed to affect the VLDL secretion rate by the isolated perfused rat liver, suggesting that this metabolite is not involved in impaired VLDL production in diabetes and after isopropanol administration.