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Biomedical subjects

D Steinberg

Publications and source records attributed to D Steinberg.

At least 19 recordsLinked to original sources
PubMed1992

Evidence for a concerted reaction between lipid hydroperoxides and polypeptides.

The events accompanying oxidative modification of low density lipoprotein (LDL) are multiple and complex, and the precise mechanisms remain to be determined. In the present studies, we examined a simple system in which we first prepared large amounts of lipid hydroperoxides (from linoleic acid or from phospholipids containing linoleic acid) by using soybean lipoxygenase (linoleate:oxygen oxidoreductase, EC 1.13.11.12). Linoleoyl hydroperoxide was then incubated with polypeptides in the absence of metal ions. We observed the generation of fluorescent products with a spectrum like that of oxidized LDL. The generation of fluorescent products from incubation of polypeptides with linoleoyl hydroperoxide was manyfold greater than that generated on incubation with preformed 4-hydroxynonenal at the same concentration. Superoxide dismutase (superoxide: superoxide oxidoreductase, EC 1.15.1.1) had no effect on the generation of fluorescent products. Incubation of linoleoyl hydroperoxide with cytochrome c (cyt c) under the same conditions led to progressive reduction of cyt c at a rate determined by the initial linoleoyl hydroperoxide concentration. This reduction was not significantly inhibited by probucol but was inhibited, although never completely, by superoxide dismutase. Even at 100 micrograms/ml, superoxide dismutase inhibited by only 65%. From these results, we are led to suggest a concerted reaction between the peroxy radical and free amino groups of polypeptides or phosphatidylethanolamine to generate fluorescent adducts. During oxidation of LDL or of cell membranes, this mechanism may occur side by side with the conventional Schiff base mechanism.

Cytochrome c Group↗
PubMed1992

The effect of delmopinol on glucosyltransferase adsorbed on to saliva-coated hydroxyapatite.

The aim was to explore the effects of delmopinol, a substituted amino-alcohol compound recently reported as a potential antiplaque agent, on GTF activity in solution and when adsorbed on to sHA. Delmopinol was without a significant effect on GTF activity in solution. In contrast, a reduction in the bound glucans synthesized by the adsorbed GTF was found in the presence of delmopinol. Delmopinol did not displace the adsorbed GTF from the sHA, nor was there significant desorption of glucans from sHA. The total glucan synthesis (bound and unbound) was reduced in the presence of delmopinol. Inhibition of GTF was not reversed by sucrose. Inhibition of GTF activity by delmopinol apparently results from drug-enzyme interaction on the surface of sHA beads. These observations provide further support for the important differences in the properties of adsorbed GTF and GTF in solution, illustrating that GTF-drug interaction differs between enzyme adsorbed to surfaces and enzyme in solution.

Adsorption↗
PubMed1992

Microbiological evaluation of the efficacy of chlorhexidine in a sustained-release device for dentine sterilization.

The aim of this in-vitro study was to evaluate the effect of chlorhexidine in solution and in a sustained-release device as an intracanal medication. Hollow cylindrical dentine specimens prepared from bovine incisors were incubated with Streptococcus faecalis for a period of 3 weeks. The intracanal medicaments tested were 0.2% chlorhexidine gluconate solution (CH), chlorhexidine in a sustained-release device (1.2 mg) (SBD), camphorated paramonochlorophenol (CMCP), and a control (no medication). Each medicament was introduced into the lumen of one dentine specimen and incubated for 5 min, 24 h, 48 h or 7 days at 37 degrees C. The bacteriological samples were taken by shaving the dentine inside the lumen with dental burs ranging in size from ISO 023-033. The dentine powder was collected in test-tubes containing growth medium and incubated for 24 h. The optical density of the medium was recorded by means of a spectrophotometer at a wavelength of 540 nm. There was a statistically significant difference between the control group and all the medicaments tested.

Animals↗
PubMed1992

Efficacy of a sustained-release device containing chlorhexidine and Ca(OH)2 in preventing secondary infection of dentinal tubules.

This study was conducted to evaluate the efficacy of the antibacterial activity of Ca(OH)2 and a sustained-release device containing chlorhexidine (SRD) in both sterilization and prevention of secondary infection of the root canal system. Bovine root dentine specimens previously incubated with Streptococcus faecalis were used in this experiment. Two different formulations of the SRD (differing in their cross-linkage), Ca(OH)2 and normal saline (control) were evaluated. The degree of bacterial infection of the root canal was tested after incubation periods of 24 h, 72 h and 7 days with these medicaments. Their efficacy in preventing secondary infection after recontamination was tested after 72 h and 7 days. The results demonstrated that both formulations of the SRD significantly reduced the bacterial population in the primary infected groups, as well as preventing secondary infection of the dentinal tubules in the recontaminated group. By contrast, Ca(OH)2 did not show any antibacterial activity, and failed to sterilize the dentinal tubules or prevent secondary infection after recontamination at the time periods examined.

Animals↗
PubMed1992

Dental caries and its determinants among recent immigrants from rural Ethiopia.

Adults (35-45 yr old) and children (5 and 12 yr old) recently arrived in Israel from rural areas in Ethiopia were examined. Caries levels were low: e.g. DMFT = 0.31 among 12-yr-olds, and 1.27 amongst adults. The 5-yr-olds were 86.8% caries-free, while 12-yr-olds were 81.8% and adults 54% caries-free respectively. According to interview data, the diet in Ethiopia had been based on local agricultural products and was almost sugar-free. The mean total count of salivary bacteria, as determined on blood agar, was 3.4 x 10(8); mean count of Streptococcus viridans, on mitis salivarius, was 6.7 x 10(7); and mean count of S. mutans, as determined on mitis salivarius with bacitracin, was 1.7 x 10(7). These levels were all high and were not significantly different from a control group of 20 Israelis. The mean number of lactobacilli, on Rogosa agar, was 2.75 x 10(4), which was significantly higher than among the controls (3.6 x 10(3). Salivary pH levels were generally similar between the Ethiopian group and the controls. Salivary flow was significantly higher for the Ethiopians (1.93 ml/min) than for controls (1.16 ml/min). Low levels of caries in this population can be attributed to an almost sugar-free diet and high salivary flow, but not to the composition of oral microflora.

Adult↗
PubMed1992

The role of oxidized low-density lipoproteins in the pathogenesis of atherosclerosis.

Hypercholesterolemia is a major risk for atherogenesis. Recent evidence suggests that oxidative modification of the major cholesterol-carrying lipoprotein, low-density lipoprotein (LDL), renders it more atherogenic. Not only does oxidized LDL (Ox-LDL) have enhanced uptake by macrophages, which contributes directly to foam cell formation, it may also adversely affect many other aspects of arterial wall metabolism and thus contribute further to the atherogenic process. Inhibition of the oxidation of LDL may be another approach to inhibiting atherogenesis, additive to or even synergistic with lowering of plasma LDL levels.

Animals↗
PubMed1992

Monoclonal antibodies against LDL further enhance macrophage uptake of LDL aggregates.

Self-aggregates of low density lipoprotein (LDL) are taken up and degraded more rapidly by macrophages than is native LDL. That enhanced uptake is attributable in part to phagocytosis via the LDL receptor pathway. However, arterial macrophages appear to express little LDL receptor activity. The present studies demonstrate an alternative mechanism by which LDL aggregates could contribute to foam cell formation. This could occur by the formation of large immune complexes that are taken up by macrophages via the Fc receptor. When immune complexes were formed with native, soluble LDL and MB47, a monoclonal antibody specific to the apoprotein B domain recognized by the LDL receptor, the subsequent uptake and degradation of the LDL by macrophages were inhibited 50-80% compared with native LDL alone. In contrast, when aggregated LDL was bound to MB47 at a similar molar ratio, the subsequent degradation of the insoluble immune complexes was two- to fivefold greater than that of aggregated LDL alone. The enhanced uptake was abolished when Fab or F(ab')2 fragments of MB47 were substituted for the intact antibody, indicating that the increased uptake was via the Fc receptor pathway. Furthermore, the uptake of the immune complexes of aggregated LDL was reduced by competition for the Fc receptor with heat-aggregated immunoglobulin. There was also an increase in the rate of cellular cholesterol esterification and an increase in macrophage cholesteryl ester mass. Since aggregates of LDL as well as autoantibodies against modified LDL have been demonstrated in atherosclerotic lesions, it is possible that immune complexes of aggregates of modified LDL may be generated in the intima.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1992

Autoradiographic analysis of the distribution of 125I-tyramine-cellobiose-LDL in atherosclerotic lesions of the WHHL rabbit.

It is well established that plasma lipoproteins enter the artery wall and play a role in the atherogenic process. However, it is still unclear where within developing atherosclerotic lesions lipoproteins accumulate and which arterial cells participate in the metabolism of these lipoproteins. For this reason, light and electron microscopic autoradiograms were prepared from sections of lesioned aortas of Watanabe heritable hyperlipidemic (WHHL) rabbits 44 hours after injection of 125I-tyramine cellobiose-low density lipoprotein (TC-LDL). After uptake of 125I-TC-LDL and intracellular degradation of the LDL protein, the nondegradable TC ligand remains trapped and thus demarcates the cells participating in the degradation of LDL. Results of other studies indicate that 48 hours after injection into WHHL rabbits, about one half of the 125I label present in lesions represents accumulated degradation products while the remaining 125I label is present as intact 125I-TC-LDL. The distribution of autoradiographic silver grains was analyzed at low resolution in fatty streaks, transitional lesions, and advanced atheroma. In all cases, the majority of silver grains were associated with superficially located subendothelial macrophage-derived foam cells. In more advanced lesions, labeling was predominant in foam cells situated within the lateral margins of the lesions. Morphometric quantification of the distribution of silver grains in electron photomicrographs of fatty streaks from two young WHHL rabbits strongly supported the data obtained at the light microscopic level. In early fatty streaks from the aortic arch and the thoracic and abdominal aortas, subendothelial macrophage-derived foam cells contained a high proportion of the silver grains (40-60% of the total) and accounted for between 30% and 40% of the lesion volume. In contrast, smooth muscle cells in the lesions contained only 7-10% of the total silver grains and accounted for approximately 20% of the lesion volume. Endothelial cells contained the most silver grains on a per-unit-volume basis by occupying only 1-2% of the lesion volume. However, the endothelium contained less than 5% of the total grains in lesions. The remaining silver grains (25-45%) were associated with the extracellular matrix, which constituted between 40% and 50% of the lesion volume. These data indicate that in the WHHL rabbit, subendothelial macrophage-derived foam cells avidly accumulate and metabolize LDL despite having few functional LDL receptors.

Animals↗
PubMed1992

Interactions of delmopinol with constituents of experimental pellicle.

The prolonged retention of an effective chemotherapeutic agent on oral surfaces and in dental plaque aids in plaque control. The objective of this study was to investigate interactions between delmopinol, a morpholinoethanol derivative, and experimental pellicle. Hydroxyapatite beads were coated with different constituents of pellicle (e.g., saliva, carbohydrates, cell-free enzymes, and bacteria). Delmopinol demonstrated a higher affinity for saliva-coated hydroxyapatite (sHA) and for experimental pellicle coated with in situ-synthesized glucans than for untreated hydroxyapatite. High-molecular-weight (MW) dextran but not low-MW dextran interfered with the adsorption of delmopinol to sHA. Delmopinol did not compete with dextran for the same binding sites on sHA, nor did it compete with saliva for the same binding sites on untreated hydroxyapatite. Delmopinol inhibited the activity of cell-free fructosyltransferase adsorbed onto sHA. In addition, synthesis of glucans by Streptococcus mutans adsorbed onto sHA was significantly reduced in the presence of delmopinol.

Adsorption↗
PubMed1992

Macrophage colony-stimulating factor mRNA and protein in atherosclerotic lesions of rabbits and humans.

In this study, the authors demonstrate the expression of mRNA and the presence of protein for macrophage colony-stimulating factor (MCSF) in atherosclerotic lesions from humans and rabbits. In situ hybridization of serial sections of human fatty streaks demonstrated expression of MCSF mRNA by cells dispersed throughout the lesions. Immunocytochemical staining with a panel of MCSF-specific antibodies showed extensive cell-associated staining of all of the cell types in the lesions. Immunocytochemical studies of atherosclerotic lesions from Watanabe heritable hyperlipidemic (WHHL) and cholesterol-fed rabbits demonstrated a similar cell-associated pattern of staining. There was no MCSF-specific staining of aortas from normal rabbits or of cultured aortic smooth muscle cells from either humans or rabbits. Macrophage-derived foam cells (MFC) were isolated from the aortas of ballooned, cholesterol-fed rabbits. A Northern blot demonstrated that RNA isolated from the MFC hybridized with a human cDNA probe for MCSF. RNA from alveolar macrophages isolated simultaneously from the same rabbits did not hybridize with the MCSF probe. Conditioned media from an 18- to 24-hour incubation of the MFC contained colony-stimulating activity as demonstrated in a mouse bone marrow culture assay. Most of this colony-stimulating activity was neutralized by preincubating the conditioned media with an MCSF-specific antibody.

Animals↗
PubMed1991

Macrophages and smooth muscle cells express lipoprotein lipase in human and rabbit atherosclerotic lesions.

Lipoprotein lipase (LPL; EC 3.1.1.34) may promote atherogenesis by producing remnant lipoproteins on the endothelial surface and by acting on lipoproteins in the artery wall. In vitro, smooth muscle cells and macrophages synthesize LPL, but in human carotid lesions only a few smooth muscle cells were reported to contain LPL protein. Endothelial cells do not synthesize LPL in vitro, but in normal arteries intense immunostaining for LPL is present on the endothelium. We used Northern blot analysis, in situ hybridization, and immunocytochemistry of human and rabbit arteries to determine cellular distribution and the site of the synthesis of LPL in atherosclerotic lesions. Northern blot analysis showed that LPL mRNA was detectable in macrophage-derived foam cells isolated from arterial lesions of "ballooned" cholesterol-fed rabbits. In situ hybridization studies of atherosclerotic lesions with an antisense riboprobe showed a strong hybridization signal for LPL mRNA in some, but not all, lesion macrophages, which were mostly located in the subendothelial and edge areas of the lesions. Also, some smooth muscle cells in lesion areas also expressed LPL mRNA. Immunocytochemistry of frozen sections of rabbit lesions with a monoclonal antibody to human milk LPL showed intense staining for LPL protein in macrophage-rich intimal lesions. The results suggest that lesion macrophages and macrophage-derived foam cells express LPL mRNA and protein. Some smooth muscle cells in the lesion areas also synthesize LPL. These data are consistent with an important role for LPL in atherogenesis.

Animals↗
PubMed1991

Expression of monocyte chemoattractant protein 1 in macrophage-rich areas of human and rabbit atherosclerotic lesions.

The recruitment of monocyte-macrophages into the artery wall is one of the earliest events in the pathogenesis of atherosclerosis. Monocyte chemoattractant protein 1 (MCP-1) is a potent monocyte chemoattractant secreted by many cells in vitro, including vascular smooth muscle and endothelial cells. To test whether it is expressed in the artery in vivo, we used Northern blot analysis, in situ hybridization, and immunocytochemistry to study the expression of MCP-1 in normal and atherosclerotic human and rabbit arteries. Northern blot analysis showed that MCP-1 mRNA could be isolated from rabbit atherosclerotic lesions but not from the intima media of normal animals. Furthermore, MCP-1 mRNA was extracted from macrophage-derived foam cells isolated from arterial lesions of ballooned cholesterol-fed rabbits, whereas alveolar macrophages isolated simultaneously from the same rabbits did not express MCP-1 mRNA. MCP-1 mRNA was detected by in situ hybridization in macrophage-rich regions of both human and rabbit atherosclerotic lesions. No MCP-1 mRNA was found in sublesional medial smooth muscle cells or in normal arteries. By using immunocytochemistry, MCP-1 protein was demonstrated in human lesions, again only in macrophage-rich regions. Immunostaining of the serial sections with an antiserum against malondialdehyde-modified low density lipoprotein indicated the presence of oxidized low density lipoprotein indicated the presence of oxidized low density lipoprotein and/or other oxidation-specific lipid-protein adducts in the same areas that contained macrophages and MCP-1. We conclude that (i) MCP-1 is strongly expressed in a small subset of cells in macrophage-rich regions of human and rabbit atherosclerotic lesions and (ii) MCP-1 may, therefore, play an important role in the ongoing recruitment of monocyte-macrophages into developing lesions in vivo.

Adult↗
PubMed1991

Alcohol and atherosclerosis.

Considerable progress has been made in recent years in our understanding of atherogenesis and, in particular, of how it relates to lipoprotein metabolism. In this conference, we attempt to re-evaluate the data on the relation between alcohol intake and coronary heart disease, emphasizing the effects of alcohol on lipoprotein metabolism. Epidemiologic data generally show an inverse correlation between coronary heart disease risk and moderate alcohol intake (variously defined but generally corresponding to 2 to 4 drinks per day). The potentially drastic effects of excessive alcohol intake on health, however, preclude any recommendation that patients increase their alcohol consumption. Equally, there may be no basis for prescribing moderate alcohol intake but, even here, the data are far from complete. The mechanism by which moderate alcohol intake "protects" remains unclear. Perhaps the best available hypothesis relates to the increased concentration of high-density lipoprotein (HDL) cholesterol associated with moderate alcohol intake. However, it should be stressed that we are still uncertain about the mechanisms linking a high HDL level to protection against coronary heart disease. If a high HDL level is only a marker (and not directly protective), raising HDL levels need not confer protection. Alcoholism, on the other hand, is associated with marked elevation of triglyceride-rich lipoproteins and is among the most common causes of hypertriglyceridemia. Moreover, once hepatic damage occurs, plasma HDL levels may actually be lower than normal. The determining point is that high alcohol intakes are associated with increased overall mortality. Until we know more about the metabolic and behavioral effects of alcohol and about its linkage to atherosclerosis, we have no basis for recommending either that patients increase their alcohol intake or that they start to drink if they do not already.

Alcohol Drinking↗
PubMed1991

Fibroblast chemotaxis after tendon repair.

Healing canine flexor tendons were treated with early controlled passive mobilization. The repair site and proximal and distal tendon stumps were stained for fibronectin and examined by light microscopy at three, seven, eleven, and seventeen days. Fibronectin increased dramatically in the epitenon adjacent to the repair site seven days after repair, a time when epitenon cellular activity was at its peak. By seventeen days, fibronectin staining had decreased substantially, both at the repair site and in the tendon stumps. A delayed increase in fibronectin activity was noted in the endotenon adjacent to the repair site. Fibronectin production appears to be an important component of the early tendon repair process. Fibroblast chemotaxis and adherence to the substratum in the days after injury and repair appears to be related directly to fibronectin secretion. This study is the first to provide documentation of fibronectin localization in a clinically relevant tendon repair model.

Animals↗
PubMed1991

Feasibility of using an oleate-rich diet to reduce the susceptibility of low-density lipoprotein to oxidative modification in humans.

Oxidized low-density lipoprotein (LDL) is more atherogenic than native LDL. The initial step in the oxidation is the peroxidation of polyunsaturated fatty acids. Thus, decreasing the concentration of polyunsaturated fatty acids should reduce the susceptibility of LDL to oxidation. Therefore, we tested the possibility that diets enriched in oleate might result in LDL that is less susceptible to oxidative modification. LDL isolated from subjects consuming an oleate-enriched diet, compared with LDL from subjects on a linoleate-enriched diet, contained significantly more oleate (28.7% vs 11.5%) and less linoleate (31.9% vs 50.9%). Generation of conjugated dienes was significantly lower in the LDL from the oleate group. Most important, after incubation with endothelial cells, LDL from the oleate group underwent less degradation by macrophages. These studies demonstrate the feasibility of altering the diet in a way that will not raise LDL cholesterol concentrations and yet will decrease the susceptibility of LDL to oxidative modification.

Adult↗