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Biomedical subjects

J Polonovski

Publications and source records attributed to J Polonovski.

At least 37 records · Page 2Linked to original sources
PubMed1988

Cholesterol prevents the teratogenic action of AY 9944: importance of the timing of cholesterol supplementation to rats.

These studies were conducted to determine whether dietary cholesterol supplementation could prevent fetal malformations induced by the amphipathic drug AY 9944, which is well known as a cholesterol biosynthesis inhibitor, and to investigate whether the plasma maternal sterol level and the nature of the sterols found in treated Wistar rats could explain this prevention. Pituitary agenesis was the most constant element of holoprosencephaly when AY 9944 was administered on d 4 of gestation at two dosages, 50 or 75 mg/kg. The rate of malformed fetuses was dose related. A strong negative correlation was established between maternal plasma sterol levels on d 10 of gestation (day of pituitary gland formation) and the rate of fetal anomalies (r = -0.97, P less than 0.01). Supplementation of AY 9944-treated rats with cholesterol had an obvious preventive action on fetal malformations. When cholesterol was added to the diet the same day as AY 9944 treatment and maintained until d 15, the prevention of malformations was almost complete. When the supplementation was initiated later, the prevention of anomalies decreased. The nature of plasma maternal sterols shows that the cholesterol supplementation modifies significantly the ratio of cholesterol to 7-dehydrocholesterol in treated rats. Therefore, maternal plasma sterol perturbations may play a role in the teratogenic action of AY 9944.

Abnormalities, Drug-Induced↗
PubMed1988

Effects of the antithrombotic drug suloctidil on low density lipoprotein processing and cholesterol metabolism in cultured human fibroblasts.

Human foetal lung fibroblasts were pretreated for 24 h with the antithrombotic drug, suloctidil (1 to 10 mumol/l), which induced a dose-dependent increase in LDL binding, uptake and degradation. At 10 mumol/l suloctidil, the respective increases in these parameters were 40%, 80% and 50%. The same treatment also resulted in increases of 1.5 to 2-fold in the synthesis of sterols, fatty acids and triacylglycerols from sodium acetate. In contrast, the esterification of cholesterol with oleic acid was specifically decreased by 35% by 24 h pretreatment of fibroblasts with 10 mumol/l suloctidil. A similar decrease of cholesterol esterification was observed in cholesterol-laden fibroblasts. It is suggested that these effects of suloctidil on LDL processing and cholesterol metabolism are related to the amphiphilic characteristics of the drug and to its calcium-blocking properties.

Cell Line↗
PubMed1988

Phenothiazines inhibit cholesteryl ester formation in J 774 monocyte-like cells.

The effect of phenothiazines (trifluoperazine and chlorpromazine) on cholesteryl ester metabolism has been investigated in J 774 mouse monocyte-macrophages. The incorporation of oleic acid into cholesteryl ester and the activity of acylcoenzyme A: cholesterol-O-acyltransferase were strongly decreased in cells pretreated for 24 h with trifluoperazine or chlorpromazine. Furthermore, trifluoperazine or chlorpromazine decreased the degradation of acetylated low density lipoprotein by J 774 cells. When cell homogenates were preincubated in vitro with trifluoperazine or chlorpromazine, a marked inhibition of acylcoenzyme A: cholesterol-O-acyltransferase activity was observed. In cells incubated with acetylated low density lipoprotein loaded with radiolabeled cholesteryl-linoleate, trifluoperazine and chlorpromazine dramatically reduced the radioactivity recovered in cholesteryl esters. The radioactivity recovered in free cholesterol was also decreased, but to a lesser extent. These results suggest that phenothiazines could efficiently antagonize cholesteryl ester accumulation in macrophages by at least two different mechanisms: a reduction of modified LDL catabolism, and a direct inhibition of the enzyme acylcoenzyme A: cholesterol-O-acyltransferase.

Animals↗
PubMed1988

Competitive enzyme inhibition immunoassay of apolipoprotein A-I: use of monoclonal antibodies.

An original competitive enzyme inhibition immunoassay has been developed for determination of total apolipoprotein A-I (apo A-I) in serum and in plasma. This specific assay involves a single monoclonal antibody (F59 4A12 2F4, directed to the -COOH terminal region of the apo A-I molecule) and a stable secondary plasma standard. Delipidation of serum samples exposed no additional antigenic sites, which suggests that all the apo A-I molecules express the epitope detected by Mab 4A12 on the surface of the apo A-I-containing particles. Within- and between-run CVs were, respectively, 6.1% and 7.5% at a 1.47 g/L concentration of apo A-I, 7.1% and 8.2% at 1.21 g/L, and 6.5% and 7.3% at 1.80 g/L. Results (y) correlated with those obtained with an electroimmunoassay (x), in which we used a mixture of three monoclonal antibodies, including Mab 4A12, as follows: y = 250x - 7, r = 0.744, P less than 0.001.

Antibodies, Monoclonal↗
PubMed1987

Arachidonic acid transfer from diacyl-sn-glycero-3-phosphocholine to ether phospholipids in rat peripheral blood lymphocytes.

The main phospholipid fractions of rat peripheral blood lymphocytes consisted of 30.8% diacylglycerophosphocholine and 20.3% diacylglycerophosphoethanolamine; the proportion of the ether phospholipids were small especially of alkyl- and alkenyl-glycerophosphoethanolamine (3.3 and 6.6% respectively). Other classes of phospholipids included glycerophosphoserine (10.5%), glycerophosphoinositol (6.5%) and sphingomyelin (13.6%). The incorporation of labeled arachidonic acid in the glycerophospholipids showed a rapid uptake into diacyl and ether classes of phospholipids and into triglycerides. When labeled cells were reincubated without arachidonic acid the diacylglycerophosphocholine rapidly lost its radioactivity, but a concomitant increase was observed in alkylacylglycerophosphocholine and more especially in alkenylacylglycerophosphoethanolamine and a smaller increase in alkyl- and diacyl-glycerophosphoethanolamine. The radioactivity was fairly constant in glycerophosphoserine and in glycerophosphoinositol, but decreased slightly in triglycerides. The arachidonic acid transfer reaction was completed after 2-3 hours reincubation. When labeled cells were reincubated in the presence of sodium cholate, the loss of radioactivity in the diacylglycerophosphocholine fraction increased. The transfer of arachidonic acid to ether phospholipids was the same with or without sodium cholate, but the later enhanced arachidonic acid transfer to diacylglycerophosphoethanolamine.

Animals↗
PubMed1987

Immunological detection of low-density lipoproteins modified by malondialdehyde in vitro or in vivo.

We describe an ELISA technique able to recognize malondialdehyde-modified low-density lipoproteins (LDL). For this purpose we produced antibodies to malondialdehyde-LDL, specific for the malondialdehyde modification of LDL; these antibodies recognized essentially malondialdehyde-LDL. Coating ELISA plates with the antibodies to malondialdehyde-LDL and using peroxidase-labelled antibodies to LDL, which reveal only apolipoprotein B, we obtained an accurate method of detecting malondialdehyde-modified apolipoprotein B. Preliminary studies demonstrated that this method allows the detection of lipoproteins containing malondialdehyde-modified apolipoprotein B in the serum of patients with cardiovascular diseases.

Animals↗
PubMed1987

Aspirin induces alterations in low-density lipoprotein and decreases its catabolism by cultured human fibroblasts.

Aspirin interacts in vitro with human low-density lipoprotein (LDL), which results in a decrease in free amino groups of apolipoprotein B and an increase of electrophoretic mobility of the particle. The aspirin-treated LDL was less efficiently recognized than native LDL by the apo B/E receptor of fibroblasts. These results suggest that aspirin in long-term treatment could influence the LDL-receptor pathway. However, aspirin-treated LDL did not bind to the scavenger receptor of macrophages.

Animals↗
PubMed1987

Monoclonal antibodies to human apolipoprotein A-I: characterization and application as structural probes for apolipoprotein A-I and high density lipoprotein.

Three mouse monoclonal antibodies (Mabs) to human apo A-I were produced using apolipoprotein A-I or HDL3 as immunogens. These monoclonal antibodies, 2G11, 4A12 and 4B11, were characterized for their reactivity with isolated apolipoprotein A-I and HDL in solution. The immunoblotting patterns of the HDL3 two-dimensional electrophoresis show that these three monoclonal antibodies reacted with all the polymorphic forms of apolipoprotein A-I. Cotitration experiments indicated that they correspond to three distinct epitopes. In order to locate these three antigenic determinants on the isolated apolipoprotein A-I, the reactivity of the three monoclonal antibodies has been studied on CNBr-cleaved apolipoprotein A-I. The monoclonal antibodies 2G11 and 4A12 addressed to the amino (CNBr 1) and carboxy (CNBr 4) terminal segments, respectively. In comparison with the monoclonal antibodies characterized by Weech et al. ((1985) Biochim. Biophys. Acta 835, 390-401), monoclonal antibody 4A12 is the only one described in the literature which is specific of the carboxy terminal segment of apolipoprotein A-I. Monoclonal antibody 4B11 does not react with any CNBr fragment, its binding is temperature dependent, it could be directed to a conformational epitope. Relative differences were demonstrated in the expression of the three epitopes in HDL subfractions isolated by density gradient ultracentrifugation. According to Curtiss and Edgington ((1985) J. Biol. Chem. 260, 2982-2993) our results indicate the existence of an immunochemical heterogeneity in the organization of apolipoprotein A-I at the surface of HDL particles as well as in the soluble form of apolipoprotein A-I.

Antibodies, Monoclonal↗
PubMed1987

Comparative study of the effect of beta-blockers with different pharmacological properties on cholesteryl ester formation in mouse peritoneal macrophages.

The effect of three beta-blockers: non-selective (propranolol), beta 1-selective (metoprolol), and with intrinsic sympathomimetic activity (pindolol), was investigated on 14C-oleic acid incorporation into cholesteryl esters in mouse peritoneal macrophages. Incorporation of 14C-oleic acid into cholesteryl esters was reduced about 10-fold by propranolol at 10(-4) M while incorporation into triacylglycerols was only 30% decreased at the same concentration. Metoprolol and pindolol had no significant effect on 14C-oleic incorporation into cholesteryl esters or triacylglycerols. Finally, propranolol inhibited the acyl-coenzyme A: cholesterol-O-acyltransferase activity, measured in vitro on macrophages homogenates, while the other studied beta-blockers were ineffective. These results suggest that propranolol could antagonize cholesteryl ester accumulation by macrophages, one of the main processes involved in atherogenesis.

Adrenergic beta-Antagonists↗
PubMed1987

Effect of AY 9944 and chlorpromazine on Concanavalin A-induced stimulation of human lymphocytes.

Amphiphilic molecules AY 9944 and chlorpromazine (CPZ) inhibited DNA synthesis in Concanavalin A-stimulated lymphocytes in a dose-dependent manner. While AY 9944 strongly decreased 7-dehydrocholesterol conversion to cholesterol, CPZ did not significantly affect this reaction. Moreover, the inhibitory effect of AY 9944 and CPZ on DNA synthesis took place in the presence of cholesterol in the culture medium. These findings suggest that the mechanism of inhibition of DNA synthesis by AY 9944 or CPZ is not related to endogenous cholesterol synthesis or exogenous cholesterol supply. Results are discussed in relation to the amphiphilic properties of AY 9944 and CPZ and to the interaction of these drugs with membranes or other intracellular targets such as calmodulin.

Autoradiography↗
PubMed1987

Dibutyryl-cyclic AMP inhibits cholesterol esterification in J 774 monocyte-like cells.

The effect of dibutyryl-cyclic AMP (dbcAMP) and theophylline was investigated on oleic acid incorporation into cholesteryl esters and triacylglycerols in the mouse monocyte-macrophage cell line J 774. 24h pretreatment of macrophages with dbcAMP decreased cholesteryl ester formation in a dose-dependent manner (about 4 fold reduction for dbcAMP 10(-4)M + theophylline 10(-3)M), while oleic acid incorporation into triacylglycerols was markedly (2 to 3 fold) enhanced. The catabolism of acetylated LDL was only slightly affected (about 15-20% reduction with dbcAMP 5 X 10(-4)M + theophylline 10(-3)M). Acyl Coenzyme A: cholesterol-O-acyl-transferase activity, measured in vitro on cell homogenates, was reduced in dbcAMP-treated cells, whereas diacylglycerol acyltransferase activity was increased. These results suggest that cyclic AMP can modulate cholesteryl ester and triacylglycerol formation in macrophages, and that these metabolisms are inversely regulated. Agents which increase cyclic AMP intracellular level could be of interest for reducing cholesteryl ester accumulation in macrophages.

Acyltransferases↗
PubMed1987

Alterations in cholesterol metabolism in cultured fibroblasts from patients with Niemann-Pick disease type C.

Cholesterol synthesis, esterification and efflux were investigated in cultured fibroblasts from patients with Niemann-Pick disease type C. Sterol synthesis from sodium acetate was markedly increased in the two Niemann-Pick disease type C strains as compared to controls, either in the presence or absence of exogenous cholesterol supply by low-density lipoproteins. By contrast, cholesterol esterification was about 2-3-fold reduced when measured by oleic acid incorporation into cholesteryl esters and 10-15-fold reduced when measured with labelled free cholesterol as precursor, although acylcoenzyme-A:cholesterol acyltransferase activity was normal when studied in vitro on cell homogenates. Chase experiments with 14C-cholesterol demonstrated that the rate of cholesterol efflux was decreased by about 3-4-fold in fibroblasts from patients with Niemann-Pick disease type C. These results provide further evidence for alterations of sterol metabolism in Niemann-Pick disease type C and support the hypothesis of a trapping of exogenous cholesterol, which cannot enter the regulatory intracellular pools.

Autoradiography↗
PubMed1987

Rapid analysis of cellular lipids without extraction.

A method is described where cell suspension obtained by scraping of monolayers was directly applied on silica gel plates. Extraction and separation of different lipid classes were simultaneously obtained during chromatography. In the range of validity of the method (no more than 80 micrograms of cellular protein tested for neutral lipids and 30 micrograms for phospholipids), this technique allows rapid lipid analysis of small samples of cultured cells, bypassing all the time- and solvent-consuming extraction and evaporation steps. The method appears to be also suitable for measurement of enzymes of lipid metabolism such as acyl coenzyme A-cholesterol-acyltransferase.

Animals↗
PubMed1987

[Phospholipid methylation in fibroblasts of patients with trisomy 21].

Comparative studies of phospholipid methylation in normal and Down syndrome (trisomy 21) fibroblasts were performed on a series of 7 pairs. Two patients showed an increased phospholipid methylation pathway as compared to the appropriate controls; one patient showed a decreased pathway; and in four patients there was no significant difference. The phospholipid methylation pathway appears to be a minor pathway, as compared to the choline pathway of phosphatidylcholine synthesis and thus does not allow the detection of an eventual disturbance of monocarbon metabolism in Down syndrome.

Cells, Cultured↗
PubMed1987

Effects of oxygen-derived free radicals on type II pneumocytes in primary culture.

After a brief review of oxygen-derived free radicals formation, cellular antioxidant equipment, and in vivo oxidant lung injury, the model of type II pneumocytes in primary culture is described with its characteristics and limitations. This model has been used to study either the in vivo or the in vitro effects of free radicals: the resistance of type II cells isolated from animals exposed to oxidants may be explained by their higher level of anti-oxidant enzymes. In contrast, type II cells isolated from control animals do not develop a resistance to the toxic effects on an in vitro exposure to free radicals. This discrepancy might be the result of a possible interaction of alveolar macrophages or fibroblasts with type II cells, which remains to be explored.

Adaptation, Physiological↗
PubMed1986

In vitro interaction of the photoactive anticancer porphyrin derivative photofrin II with low density lipoprotein, and its delivery to cultured human fibroblasts.

Low density lipoprotein (LDL) doped with the anticancer mixture of hematoporphyrin derivatives Photofrin II (P2) competes with native LDL for binding to fibroblast receptors, despite a slight increase in the negative net charge related to the presence of acidic residues of porphyrins. P2 delivery to fibroblasts can be achieved by LDL, HDL3 or albumin doped with P2 (LDL-P2, HDL-P2 or A-P2, respectively). P2 delivery to cells assessed by fluorescence measurement, is much more efficient, at low protein concentrations (10-20 micrograms/ml) by LDL-P2 than by HDL-P2 or A-P2. Moreover, P2 delivery to cells by LDL-P2 as a function of protein concentration is a saturable process, whereas P2 delivery by HDL-P2 or A-P2 is a linear process. Finally, reduction of the LDL-receptor number by preincubation of fibroblasts in medium supplemented with lipoproteins results in a decrease of P2 delivery by LDL-P2. These results suggest a special role of the LDL-receptor pathway in P2 delivery to cells and could be of interest in cancer phototherapy by porphyrins.

Antineoplastic Agents↗
PubMed1986

Effect of manipulation of phospholipid polar head group on low density lipoprotein metabolism in human cultured fibroblasts.

Modification of phospholipid polar head group was achieved by growing human cultured fibroblasts in medium devoid of serum and supplemented with N-methyl ethanolamine or N,N-dimethylethanolamine during 48 h. The corresponding phospholipids accounted for approximately 45% of total phospholipids. Whereas low density lipoprotein (LDL) binding was unaffected, LDL internalization and degradation appeared to be markedly reduced in the presence of N-methylethanolamine. N,N-dimethylethanolamine had no effect on the three studied parameters. These results emphasize the importance of phospholipid polar head group in LDL processing by receptor-mediated endocytosis.

Cells, Cultured↗