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D S Goodman

Publications and source records attributed to D S Goodman.

At least 109 records · Page 6Linked to original sources
PubMed1981

Regulation of retinol-binding protein metabolism by glucocorticoid hormones in cultured H4II EC3 liver cells.

Studies were conducted to explore the effects of glucocorticoid hormones on the regulation of the metabolism of retinol-binding protein (RBP) by H4II EC4 rat hepatoma cells in culture. Cortisol, corticosterone, and the synthetic glucocorticoid analog dexamethasone all induced a 2- to 3-fold increase in accumulation of RBP. Half-maximal stimulation occurred at concentrations of dexamethasone in the range of 1-5 nM. Progesterone in the concentration range of 1-10 microM, inhibited the stimulatory effect of dexamethasone. Progesterone alone in this concentration range had no effect on RBP metabolism. By analogy with the studies of others, these observations with progesterone suggest that glucocorticoid receptors are involved in the effect of dexamethasone on RBP. As previously reported, RBP accumulated in the hepatoma cells when they were incubated in a medium free of serum and of vitamin A. The addition of retinol over a range from 3.5 nM to 3.5 microM stimulated a dose-dependent secretion of RBP from the cells into the medium. In longer experiments, retinol also stimulated the accumulation of RBP. Neither dexamethasone nor retinol had an effect on the accumulation or the cell to medium distribution of rat serum albumin or prealbumin at concentrations which were maximally stimulatory for RBP. When studied over a wide range of concentrations, retinol and dexamethasone incubated together produced approximately additive increases in the accumulation of RBP. Dexamethasone, moreover, did not affect the retinol-induced secretion of RBP. Thus, retinol and dexamethasone appear to function via different and independent mechanisms to regulate the metabolism of RBP by the liver cell.

Animals↗
PubMed1981

On determining the extent of side-pool synthesis in a three-pool model for whole body cholesterol kinetics.

Whole body cholesterol turnover is well described by a three-pool model. This model has eight unknown parameters: three masses, three synthesis rates, and two intercompartmental exchange rates. Only six parameters can be estimated by fitting the model to the plasma specific radioactivity-time curve which results from the intravenous injection of labeled cholesterol. Additional information is obtained if a precursor of cholesterol, labeled with a different isotope, is also injected. Equations are derived to enable the calculation of all eight model parameters from the two sum-of-exponentials equations that are fitted to the two tracer curves. The characteristics of a satisfactory precursor are discussed.

Animals↗
PubMed1980

Effects of vitamin A deficiency on the levels and distribution of retinol-binding protein and marker enzymes in homogenates and Golgi-rich fractions of rat liver.

The levels of retinol-binding protein, prealbumin, and several 'marker' enzymes were determined in homogenates, crude subcellular fractions and isolated Golgi apparatus prepared from the livers of vitamin A-deficient and control rats. Vitamin A deficiency led to a marked increase (3.5-fold) in hepatic retinol-binding protein concentration and to slight increases in hepatic pre-albumin levels, without affecting the levels of a number of marker enzymes localized in various subcellular compartments. The distributions of total protein and marker enzymes among various subcellular fractions were nearly identical in control and vitamin A-deficient preparations. In particular, vitamin A deficiency had no effect on the yield or enzymatic composition of isolated Golgi-rich fractions. In vitamin A-deficient rats, where the normal secretion of retinol-binding protein was blocked, a maximum of less than 10% of the total liver retinol-binding protein was accounted for in the Golgi. In contrast, in control rats, where the secretion of retinol-binding protein was proceeding at the normal rate, the relative amount of retinol-binding protein in Golgi increased to about 23% of the total liver pool. The data suggest that the Golgi apparatus is involved in the pathway of retinol-binding protein secretion from the liver, but demonstrate that the Golgi is not the major subcellular locus for retinol-binding protein in either normal or vitamin A-deficient rats.

Animals↗
PubMed1980

Plasma retinol-binding protein.

Vitamin A is mobilized from liver stores and transported in plasma in the form of the lipid alcohol retinol, bound to a specific transport protein, retinol-binding protein (RBP). A great deal is known about the chemical structure, metabolism, and biological roles of RBP. RBP is a single polypeptide chain with molecular weight close to 20,000. RBP interacts strongly with plasma prealbumin, and normally circulates in plasma as a 1:1 molar RBP-prealbumin complex. Both the primary and the tertiary structure of prealbumin are known, and the primary structure of RBP has recently been reported. Much information is available about the protein-protein and protein-ligand interactions that are involved in this transport system. Many clinical studies have examined the effects of a variety of diseases on the plasma levels of RBP and prealbumin in humans. Plasma RBP levels are low in patients with liver disease and are high in patients with chronic renal disease. These findings reflect the facts that RBP is produced in the liver and mainly catabolized in the kidneys. Delivery of retinol to extra-hepatic tissues appears to involve specific cell surface receptors for RBP. Vitamin A mobilization from the liver, and delivery to peripheral tissues, is highly regulated by factors that control the rates of RBP production and secretion. Retinol deficiency specifically blocks the secretion of RBP, so that plasma RBP levels fall and liver RBP levels rise. Injection of retinol into vitamin A-deficient rats stimulates the rapid secretion of RBP from the liver into the plasma. The cellular and molecular mechanisms that mediate these phenomena are under investigation. Elucidation of these mechanisms should help define the basic mechanisms that control the mobilization, transport, and delivery of vitamin A.

Animals↗
PubMed1980

Origin of platelet-derived growth factor in megakaryocytes in guinea pigs.

Growth factor activity, as determined by the stimulation of [3H]thymidine incorporation into the DNA of quiescent 3T3 cells in culture, was found in lysates of guinea pig platelets and megakaryocytes. Quantitative dilution studies demonstrated that, of the cells present in the guinea pig bone marrow, only the megakaryocyte possessed quantitatively significant growth factor activity. The amount of activity present in one megakaryocyte was equivalent to that present in 1,000-5,000 platelets, a value approximately comparable to the number of platelets shed from a single megakaryocyte. It is suggested that guinea pig platelet-derived growth factor has its origin in the megakaryocyte.

Animals↗
PubMed1980

The binding protein for retinoic acid from rat testis cytosol: isolation and partial characterization.

This study reports the isolation and partial characterization of a soluble protein with binding specificity for retinoic acid from rat testis cytosol. Cytosol, labeled in vitro by incubation with [14C]retinoic acid, was fractionated by a series of procedures that included ion exchange chromatography on DEAE-Sepharose and on DEAE-cellulose, gel filtration on Sephadex (G-50, and preparative polyacrylamide gel electrophoresis. The resulting cytosol bindin; protein for retinoic acid had been purified approximately 16,000-fold, with recovery of 11% of the specifically bound [14C]retinoic acid. The purified binding protein was homogeneous on disc gel electrophoresis at pH 4.5 or in the presence of SDS, but displayed microheterogeneity on isoelectric focusing, where both a major and a minor band with apparent isoelectric points near 4.7 were observed. Purified retinoic acid binding protein had a molecular weight of approximately 14,600, and an ultraviolet absorption spectrum with two maxima near 277 and 346 nm. The fluorescence intensity of retinoic acid bound to the cytosol binding protein was approximately 30-times greater than that of retinoic acid in solution in any of five organic solvents, suggesting that retinoic acid is highly immobilized when bound to the cytosol binding protein. The properties of the rat testis cytosol binding protein for retinoic acid were compared directly with those of the testis cytosol binding protein for retinol and of serum retinol-binding protein. The two cytosol binding proteins were much more similar to each other than to the serum transport protein, from which they differed in a number of properties, including which they differed in a number of properties, including molecular size, affinity for prealbumin, immunological reactivity, and absorption and fluorescence spectral characteristics.

Amino Acids↗
PubMed1980

Colchicine inhibition of retinol-binding protein secretion by rat liver.

Studies were conducted to explore the effects of colchicine on the secretion and metabolism of retinol-binding protein (RBP) by the liver. In the vitamin A-deficient rat, the rate of secretion of RBP from the liver into the serum is greatly reduced, and RBP accumulates in the liver. Injection of retinol (dispersed in a 20% Tween 40 solution) into deficient rats stimulated a rapid secretion of RBP from the liver into the serum. Colchicine treatment markedly inhibited the retinol-stimulated secretion of RBP from the liver into the serum. The effect of colchicine was most pronounced during the early period after retinol injection, particularly during the first 30 to 60 minutes. Ninety minutes after retinol injection, the serum RBP level of colchicine treated rats was only 36% as great as that of the control rats. In parallel experiments, a quantitatively similar inhibition of very low density lipoprotein (VLDL) secretion by colchicine was observed in the retinol-deficient rats. In contrast, colchicine did not affect the overall rate of hepatic protein synthesis, as estimated from the incorporation of [(3)H]leucine into total liver and serum protein; the secretion of newly synthesized protein was, however, inhibited. When rats were treated with colchicine and then injected with retinol, the level of RBP in a Golgi-rich fraction obtained from the liver homogenate increased markedly as compared to the level of prealbumin. The inhibition of RBP secretion by colchicine suggests that the microtubules play a role in RBP secretion. By analogy to studies on VLDL and albumin, these data provide presumptive evidence that the Golgi apparatus and secretory vesicles are involved in RBP secretion.-Smith, J. E., D. D. Deen, Jr., D. Sklan, and D. S. Goodman. Colchicine inhibition of retinol-binding protein secretion by rat liver.

Animals↗
PubMed1980

Prediction of the parameters of whole body cholesterol metabolism in humans.

Total body turnover of cholesterol was studied in 54 subjects by fitting a three-pool mathematical model to plasma decay curves of 32--49 weeks duration following [14C]cholesterol injection. Fifteen subjects were normal, 10 hypercholesterolemic, 21 hypertriglyceridemic, and 8 had both hypercholesterolemia and hypertriglyceridemia; 21 had a familial form of hyperlipidemia. In every subject in this heterogeneous population, the three-pool model gave the best fit for the data. An extensive search was conducted for relationships between model parameters and physiological variables (body size, serum lipid levels, age, and sex). Both linear and nonlinear relationships, and those involving interactions between pairs of variables, were explored. Fifty different forms of the model parameters and 53 forms of the physiological variables were examined. To guard against declaring statistical significance when none was present, subjects were first randomly divided into two matched groups. In the first (hypothesis-generating) group of 36 subjects, more than 100,000 regression equations were considered for each form of the model parameters. Twenty-one highly significant equations were found that were then tested in the second group (hypothesis-testing, 18 subjects). Eighteen of the 21 equations were found to be significant; of these, 6 were selected that accounted for a large part of the observed variation in the four model parameters for which equations were found (production rate (PR), and the sizes of pool 1, pool 3, and total exchangeable body cholesterol). The major determinant of cholesterol PR was body weight alone (r = 0.80). No function of serum lipid levels significantly influenced PR. Both body weight and serum cholesterol level entered into the equations for cholesterol mass. Age influenced the size of pool 3. Serum triglyceride level only had an effect on the size of pool 1. Since these equations were generated in one group of subjects and tested in another, they can be considered a confirmed set of predictive equations.

Adult↗
PubMed1979

Quantitative studies of the interaction of cholecalciferol ((vitamin D3) and its metabolites with different genetic variants of the serum binding protein for these sterols.

Cholecalciferol (vitamin D3) and its 25-hydroxy metabolite are transported in plasma bound to a specific protein, the binding protein for cholecalciferol and its metabolites (DBP). DBP is identical with the group-specific component (Gc) proteins, which are known to display genetic polymorphism. Studies were conducted to explore whether or not major differences in the transport of cholecalciferol and its biological metabolites might exist among persons with different Gc phenotypes. Detailed quantitative studies were first carried out on the interaction of 25(OH)D3 with DBP in 21 different samples of serum, representing eight different Gc phenotypes. The studies used a filter disc assay method that provided highly reproducible quantitative results with cholecalciferol-related sterols. The Gc phenotypes studied included the three common types (Gc 1-1, 2-1, and 2-2) and several uncommon genetic variants (Gc Ab-Ab, Ab-1, Ab-2, Chip-1, and Chip-2). The binding affinities for 25(OH)D3 observed with these different sera were all fairly similar to each other. More extensive studies were then conducted to compare the binding of four cholecalciferol-related sterols to each of three genetic variants of DBP, by using sera from homozygous persons with the Gc 1-1, Gc 2-2 and Gc Ab-Ab phenotypes. The ligands tested included cholecalciferol, 25(OH)D3, 1,25(OH)2D3, and 24(R) 25(OH)2D3. The affinities of the three genetic types of DBP/Gc protein were found to be similar for each of the four cholecalciferol-related sterols. The apparent association constants for 25(OH)D3 and 24,25(OH)2D3 were similar (approx. 1--2 x 10(8) M-1); lesser affinities were observed for 1,25(OH)2D3 (kA approx. 1 x 10(7) M-1) and for cholecalciferol (kA approx. 3--4 x 10(5) M-1). Thus the common genetic variants of DBP/Gc protein, and the uncommon genetic variants studied here, all appear to have similar binding properties for cholecalciferol and its several metabolites.

Alpha-Globulins↗
PubMed1979

Release of platelet-derived growth factor from human platelets by arachidonic acid.

Platelet alpha-granules contain a factor that stimulates the proliferation of arterial smooth muscle cells and may play a role in atherogenesis. We have studied the role of arachidonic acid in mediating the release of the platelet-derived growth factor (PDGF) from human platelets. PDGF was assayed by stimulating of [(3)H]thymidine incorporation into DNA of mouse 3T3 cells. Platelet aggregation and the release of platelet factor 4,beta-thromboglobulin, and serotonin were also studied. A biphasic response pattern was observed when gel-filtered platelets were incubated with arachidonate over the concentration range 0.01-0.4 mM. At low arachidonate levels (approximately 0.025-0.1 mM), specific concentration-dependent aggregation and release of PDGF and of the other components were observed. This effect was not seen with any of five other fatty acids tested and was suppressed by indomethacin (25 muM). At higher arachidonate concentrations (approximately 0.15-0.35 mM), a concentration-dependent turn-off of both aggregation and release occurred. At these concentrations the platelets remained functional, and no release of lactate dehydrogenase was observed. A similar biphasic pattern of arachidonate-induced aggregation and release was observed with platelet-rich plasma, over a similar range of arachidonate to albumin mole ratios. These studies demonstrate that PDGF and other alpha-granule constituents can be released from platelets specifically by arachidonate via an indomethacin-sensitive pathway, most probably involving the platelet cyclooxygenase and conversion of arachidonate to prostaglandin metabolities. The mechanisms responsible for the turn-off of the specific arachidonate-mediated responses at higher arachidonate concentrations remain to be defined.

Arachidonic Acids↗
PubMed1979

Normal levels of fibrinopeptide A in patients with primary hyperlipidemia.

Fibrinopeptide A (FPA) levels were measured in a group of 130 controls and patients with various types of primary hyperlipidemia to investigate whether an increased steady state level of thrombin activity is present in hyperlipidemic patients. In a subset of 56 subjects, levels of clotting factors II, VII, and X were measured as well. FPA levels in hyperlipidemic patients were not significantly different from those of control subjects. Furthermore, on multiple regression analysis, no significant relationships were found between FPA levels and the concentrations of serum cholesterol or triglyceride, or log triglyceride levels. Statistically significant relationships were found between all three clotting factor levels and triglyceride concentration. The correlation coefficients for these relationships, however, were low, so that the correlations are of questionable pathophysiological significance. A weak relationship also was found between the plasma levels of cholesterol and of factor II. Thus, although small increases in various clotting factors may be found in patients with hyperlipidemia, plasma FPA levels are normal. These data indicate that hyperlipidemia is not associated with a steady state of increased thrombin activity in vivo in humans.

Cholesterol↗
PubMed1979

Unusual properties of retinyl palmitate hydrolase activity in rat liver.

These studies report the hydrolysis of retinyl palmitate with liver homogenates and homogenate fractions from retinol-depleted rats. The studies utilized an effective in vitro assay for retinyl palmitate hydrolase (RPH) activity, in which microgram amounts of retinyl palmitate were employed as substrate, followed by the chromatographic separation and fluorescence assay of free and esterified retinol. RPH activity was maximal near pH 8 in Tris-maleate buffers, and required a bile salt for stimulation. Both cholate and taurocholate stimulated the reaction, whereas a number of other detergents tested were ineffective. The enzymatic activity showed an unusual subcellular distribution, with about 40% of total RPH activity recovered in the washed "nuclear" fraction (1,500 g pellet) and about 30--35% in the 105,000 g supernatant. This unusual distribution was not observed for marker constituents for plasma membranes, nuclei, mitochondria, lysosomes, Golgi apparatus, or endoplasmic reticulum. Despite its enrichment in the "nuclear" fraction, RPH activity was not enriched in purified preparations of nuclei or plasma membranes. Thus, RPH activity was not localized in any single, characterized subcellular structure. Another striking feature of the hepatic RPH activity was its extreme variability from rat to rat as assayed in vitro. Both the unusual subcellular distribution and the marked variability in activity were not observed for a variety of other hepatic ester hydrolase activities examined. Of ten lipid and nonlipid esters tested as substrates, only the hydrolytic activities against cholesteryl oleate and phytyl oleate correlated with, and partly resembled, RPH activity in these respects. The results suggest that the observed RPH activity is relatively specific for the hydrolysis of retinyl palmitate, and may therefore be significantly involved in hepatic retinyl ester metabolism.

Animals↗