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Biomedical subjects

R Rozen

Publications and source records attributed to R Rozen.

At least 127 records · Page 7Linked to original sources
PubMed1984

Regulation and expression of carbamyl phosphate synthetase I mRNA in developing rat liver and Morris hepatoma 5123D.

A cDNA clone complementary to mRNA encoding the precursor (Mr = 165,000) to the rat liver mitochondrial matrix enzyme carbamyl phosphate synthetase I (Mr = 160,000) was employed to compare relative amounts of the messenger in adult and fetal liver and in Morris hepatoma 5123D and 3924A cells. Northern blot analysis gave a size estimate for the messenger of 6,500-6,700 nucleotides. Carbamyl phosphate synthetase mRNA levels in 15-day-old fetal liver were less than 10% of adult levels; 5123D cells expressed the messenger at levels about 2-fold higher than normal adult liver, but the messenger was undetectable in 3924A cells. Albumin mRNA was also expressed in the former but not in the latter. Maintaining rats for 5 days on a diet containing 60% casein augmented the relative amount of carbamyl phosphate synthetase mRNA by about 2-fold, while a protein-free diet resulted in reduced levels of the mRNA (about 50% compared to animals on a normal diet). Finally, the pattern of hybridization of carbamyl phosphate synthetase cDNA to HindIII-digested genomic DNA showed no differences between normal liver and its corresponding hepatoma; however, a HindIII site polymorphism was observed between Buffalo and ACI rats.

Animals↗
PubMed1984

Effects of a very-low-calorie diet on adrenergic responsiveness in human adipose tissue.

Nineteen obese women were given a strictly-defined very-low-calorie diet for three weeks, and changes in adipose tissue lipolysis were studied using adrenergic drugs. The beta-receptor responsiveness was tested by isoprenaline. Before the diet, we found a negative relationship between basal value and isoprenaline-net effect on glycerol release; after the diet, its relationship disappeared, but the beta-stimulated level remained at a similar maximum. The alpha-receptor responsiveness was studied using clonidine, which significantly inhibits lipolysis; this effect was twice as high after the diet than before. Adrenaline-induced lipolysis was also studied; we found a dose-dependent response, and after a diet a negative linear relationship between basal and adrenaline net-lipolysis, so that adrenaline became antilipolytic for the high values of spontaneous lipolysis. We conclude that: (1) diet does not modify adipocyte-beta-receptor responsiveness, but it increases alpha-receptor responsiveness; (2) relationships between basal lipolysis and the lipolytic responsiveness of adrenergic receptors, which already exist with the spontaneous feeding, are enhanced by the hypocaloric diet, but in such a way that adipocyte-lipolysis remains within a given range; (3) we were unable to find any relationship between rate of weight loss and amplitude of adrenergic-receptor responsiveness.

Adipose Tissue↗
PubMed1983

Effects of glucagon on biosynthesis of the mitochondrial enzyme, carbamoyl-phosphate synthase I, in primary hepatocytes and Morris hepatoma 5123D.

When freshly-dispersed rat hepatocytes are maintained in primary monolayer cultures, they quickly lose their capacity to synthesize the urea cycle enzyme, carbamoyl-phosphate synthase. The ability to synthesize many other proteins, e.g., serum proteins including albumin, is retained. After an initial recovery period following cell isolation (24-48 h), glucagon is able to restore the ability of cultured hepatocytes to make carbamoyl-phosphate synthase. mRNA encoding the enzyme is about 4-times higher in hepatocytes maintained for 48 h in the presence of glucagon compared to hepatocytes without the hormone, as judged by in vitro translational assays. The level of carbamoyl-phosphate synthase activity expressed in transformed hepatocytes is unique to each hepatoma. Here we show that Morris hepatoma 5123D has retained such expression, and actively synthesizes the enzyme when 5123D cells are placed in monolayer cultures. Unlike normal hepatocytes, however, synthesis continues uninterrupted at a high level whether or not glucagon is present. 5123D has higher levels of translatable carbamoyl-phosphate synthase mRNA than normal liver.

Animals↗
PubMed1983

Hypertaurinuria in the C57BL/6J mouse: altered transport at the renal basolateral membrane.

The C57BL/6J mouse is an inbred strain with selective hypertaurinuria. C3H/HeJ mice (control) and C57BL/6J mice have comparable values for plasma taurine and renal creatinine clearance but significantly different fractional taurine excretion: C3H mice, 0.15 +/- 0.03; C57BL mice, 0.44 +/- 0.08 (P less than 0.01). C57BL mice fed control diet have significantly higher renal cortex taurine (11.5 +/- 0.4 mumol/g wet wt, mean +/- SE) relative to C3H mice (10.5 +/- 0.5 mumol/g, P less than 0.05); the difference is magnified when mice are fed low protein or low sulfur amino acid diet and experience renal adaptation with enhanced taurine transport at the luminal membrane (P less than 0.01). Taurine transport by purified brush border membrane vesicles is similar in the two strains fed control diet. On the other hand, renal cortex slices show the following differences: 1) net uptake of taurine at 0.11 mM is lower in C57BL slices at initial rates and at steady state; 2) the difference in uptake between strains is magnified in the presence of phlorizin, an inhibitor of taurine efflux; 3) efflux of taurine from preloaded slices is less per unit time in C57BL mice; and 4) uptake differences are specific for taurine. These findings are not accountable to differences in taurine oxidation, slice weight, or water spaces. They indicate that hypertaurinuria in the C57BL/6J strain is a function of decreased basolateral membrane permeability to taurine at physiologic concentrations.

Amino Acid Metabolism, Inborn Errors↗
PubMed1983

D-Glucosamine-induced increase of the glycerol-containing lipids in growing cultures of human malignant epithelial cells.

D-Glucosamine was found to inhibit the growth of human malignant epithelial cells SW-839, HT-29, RT-4, and SK-OV-3 in culture in a process that was associated with significant increments in glycerol-containing lipids. Each cell line had a different sensitivity to the drug, but all four cell lines shared the same features in their response, i.e., dose-dependent (at concentrations of 1, 5, and 10 mM), noncytotoxic reductions in growth (minimum 30%, maximum 70%), and simultaneous 1.5-fold to sevenfold increases in lipid contents. Cells regained their normal growth and lipid patterns when glucosamine was removed. Glucosamine did not modify the lipid contents of cells in the late phase of culture when growth was minimal.

Adenocarcinoma↗
PubMed1982

Renal transport of taurine adapts to perturbed taurine homeostasis.

Renal adaptation apparently contributes to the homeostasis of taurine, a beta-amino compound that behaves as a conserved metabolite in the mammal. We studied two strains of inbred mice: C3H/HeJ (low-taurine excreter) and C57BL/6J (high-taurine excreter due to impaired basolateral membrane permeability to taurine). Low-protein and low-sulfur amino acid diets fed for two weeks significantly decreased plasma taurine in both strains, decreased fractional taurine excretion in vivo (particularly in the C57BL strain), and increased net uptake of taurine by renal cortex slices and isolated brush-border membrane vesicles (BBMV) in vitro in both strains. Renal adaptation was less obvious in vivo in the low-taurine excreter C3H strain, but in vitro adaptation, as observed in slices and BBMV (P less than 0.01), was greater than that observed in the C57BL strain. Renal cellular taurine content fell (P less than 0.01) only in the adapted C3H strain. The in vitro adaptive response was not confined to taurine; BBMV uptake of D-glucose and L-alanine was also enhanced in the adapted state. Specificity of the stimulus for adaptation was tested with a low-phenylalanine diet; a modest adaptation was observed in vivo and in vitro but only in the C3H strain. BBMV adaptation did not correlate with blood methionine but correlated inversely with plasma taurine (r = 0.71, P less than 0.05), implying that change in extracellular taurine may be a signal for renal adaptation in taurine homeostasis in the mammal.

Amino Acids↗
PubMed1981

Very-low-density-lipoprotein secretion by isolated hepatocytes of fat-fed rats.

The very-low-density-lipoprotein secretion rate of isolated hepatocytes obtained from rats fed a high-fat diet was half that of cells from control animals. In fat-fed rats, the initial cellular uptake of [l-14C]oleate in vitro was decreased by 25%, its esterification to triacylglycerols and phospholipids by 50% and its incorporation into very-low-density-lipoprotein triacylglycerols by 70%. Exogenous oleate was not the main precursor of very-low-density lipoproteins in these animals. Lipogenesis, a minor source of very-low-density lipoproteins with the control diet in our experimental conditions, was inhibited by 84% after fat-feeding. A short-term inhibition of lipogenesis in vitro did not result in a decrease in very-low-density-lipoprotein secretion rate. The results suggest that fat-feeding decreased availability of exogenous as well as endogenous fatty acids for synthesis of very-low-density lipoproteins.

Animals↗
PubMed1980

Effect of a high-fat diet on rat very low density lipoprotein secretion.

1. Very low density lipoprotein (VLDL) secretion rates were studied on rats adapted to a high-fat diet (71% calories as lard) for 3-4 weeks, compared to control (starch-fed) rats. 2. Experiments were performed at 14.00 h, at which time all animals had the same circulating free fatty acids. Fat-fed rats presented an apparent liver stealosis, a high post-Triton chylomicron secretion, but a 40% decreased VLDL secretion. 3. Injection of [1-14C]palmitic acid showed that the tracer was incorporated less in liver triacylglycerols of the fat-fed rats, presumably because of an enhanced ketogenesis. Secretion of labelled VLDL-triacylglycerols in 1 h was diminished 5-fold, even after a correction for the lower hepatic esterification. 4. Two complementary experiments were carried out, with the following results: at 08.00 h, when serum free fatty acid concentrations were comparable in both groups of rats [5,10], post-Triton VLDL secretion was diminished by 45% in the fat-fed rats; at 20.00 h, the fat-fed rats had significantly elevated plasma free fatty acids [5,10], but their VLDL secretion was the same as in control rats. 5. So it appears that in fat-fed rats circulating free fatty acids do not stimulate VLDL secretion as expected. It is suggested that the decreased VLDL secretion with the high-fat diet may result from inhibition of hepatic lipogenesis.

Animals↗
PubMed1979

Taurine transport in renal brush-border-membrane vesicles.

Taurine transport in isolated brush-border-membrane vesicles from rat kidney is concentrative and it is driven by the Na+ gradient and transmembrane potential difference; binding is not a significant component of net uptake. The Na+-dependent component of net uptake is saturable with an apparent Km of 17 microM. The taurine-transport mechanism is selective for beta-amino compounds.

Amino Acids↗
PubMed1977

Evaluation of metabolic pathway activity in cultured skin fibroblasts and blood leukocytes.

Screening for a variety of blocked catabolic mutants can be achieved in cultured skin fibroblasts and in peripheral blood leukocytes with a simple radioisotope method that is reliable and convenient. The method measures the incorporation into trichloroacetic acid (TCA) insoluble macromolecules of a 14C-labelled intermediate in the pathway under question; incorporation of a 3H-labelled metabolite, not in the same pathway, is used as an internal control of metabolism in the cell population. The 14C/3H incorporation ratio is decreased in blocked pathways; use of the ratio method eliminates the delay required by radioautography (used in earlier adaptations of this method), the problems involved in CO2 collection, and the need to standardize cultures for cell number. This method has been used to identify cells with biochemical lesions in the oxidation of propionate, galactose, hypoxanthine and pyruvate; it has allowed us to identify a new variant of methylmalonicaciduria; we believe it can be extended to include other metabolites and pathways.

Acidosis↗
PubMed1976

[Short term effects of diets with high levels of dithiocarbamates on carbohydrate and lipid metabolism of rat liver].

The purpose of the work was to establish the eventual "metabolic toxicity" of pesticide-contaminated diets in the Rat. The liver metabolic response to various stimuli was compared in dithiocarbamate-fed animals and in non-contaminated ones. 112 weanling male Wistar CF rats were fed, during 15 days, with a demi-synthetic control diet. They were then divided into 4 lots:--the control group C, which went on to receive the same diet,--the nabame group N, the diet of which was supplemented with 275 ppm of the dithiocarbamate;--the thirame groupe R, receiving the control diet + 600 ppm thirame;--the zineb groupe Z, given the control diet + 3 600 ppm Zineb. The animals were fed with these diets during 14 days, their dithiocarbamate intake thus averaging 1/20 th of the per os LD 50/rat/day. At the end of this 2-week period, each of the 4 groups was divided into 4 sub-groups, all the animals were fasted overnight, then sacrified:--after no other treatment (sub-groups T);--30 minutes after an i.p. injection of 2.6 g/kg glucose (G);--after having been forced to walk in a restraint wheel for 50 minutes/hr during the 18 hrs of the night fast (sub-groups W);--after a 90 minutes exposure in a cold room (F). The weights of the animals, of their liver, heart, kidneys, adrenals and epididymal pads were recorded. In their liver, the following compounds were determined: water, proteins, total lipids, triglycerides, long-chain acyl-CoA, non-esterified fatty acids, total cholesterol, glycogen, glucose, alpha-glycerophosphate. The thirame rats had a lower food intake than the others and the smallest body weight, but their relative liver and kidneys weights were the highest. The nabame animals did not differ from the control ones but the zineb rats had the lightest epididymal fat pads. The primary effects of the dithiocarbamate diets on liver metabolism were apparently not the same in the 3 groups compared to the control ones: nabame and thirame increased glycogen, thirame increased the lipid compounds: long-chain actyl-CoA and triglycerides, where as zineb feeding resulted in an increase of glucose concentration and in a decrease of triglycerides and total lipids. Muscular exercise, or cold exposure, had the following effects compared to those they had in the control group: a greater glucose utilization in the nabame and thirame rats, a smaller glycogen and glucose utilization, associated with an increase of alpha-glycerophosphate, in the zineb animals. These results were considered altogether with those obtained in a previous paper by the same authors, which concerned liver ketone bodies and adenine nucleotides changes after the same experimental conditions, and it was concluded that the 3 dithiocarbamates actually had a common effect on rat metabolism: they all impaired glucose utilization by the liver. Also, fat mobilization from peripheral depots was shown to occur in the 3 experimental groups, resulting in liver fatty acid oxidation in the nabame and zineb rats, and in liver steatosis for the thirame ones...

Adipose Tissue↗
PubMed1975

[Energy metabolism in the liver of rats fed a diet contaminated with dithiocarbamates].

Young male rats, Wistar CF strain, weighing about 100 g, were fed during 14 days with a well-balanced diet, but containing either 275 p.p.m. nabame, either 600 p.p.m. thirame or 3 600 p.p.m. zinebe. The animals given the non-contaminated diet were the controls. On the evening before the experiment, they were all fasted and some of them, forced to walk during 18 hours in a restraint wheel. On the morning of the experiment, some of the rats which have not been working were placed in a cold room at + 4 degrees C, and some others were given an i.p. injection of 2,6 g glucose per kg body weight. The animals were then killed, those that received the glucose treatment 30 mn after the injection, the cold-exposed rats 90 mn after the beginning of their exposure. The redox and energy potentials of the liver tissue were determined after the enzymatic assay of the following liver metabolites : lactate, pyruvate, beta-hydroxybutyrate, acetoacetate, ATP, ADP, AMP, inorganic phosphate. The thirame group rats had the smallest body weight and the lowest food intake. All the pesticides-exposed animals has a higher liver weight than predicted by their body weight. The pesticides-containing diets decreased liver lactate concentration and the lac/pyr ratio. Thirame was the more efficient and it partly impaired the glucose induced increase of the cytoplasmic redox potential, as estimated from the variation of the lac/pyr ratio. The pesticide-containing diets also lowered the liver concentrations of beta-hydroxybutyrate, acetoacetate, and their ratio. Last the pesticides, which but slightly modified the liver contents in adenine nucleotides and inorganic phosphate in the fasting state, increased the ATP fall following cold exposure and decreased the net ATP synthesis produced by glucose administration. The thirame diet was the more efficient in our experimental conditions, the zinebe diet the least one. It was concluded in our discussion that dietary dithiocarbamates either induced a hyperthyroidic status in the animal, or acted themselves as thyroxin-like compounds, because the liver metabolism was more directed towards heat production than towards that of chemical energy available for syntheses.

Adenine Nucleotides↗
PubMed1975

[Lipid metabolism in rats fed a diet rich in various carbohydrates. I. Results after I.P. injection of lipogenic precursors].

Male rats, Wistar CF strain, weighing 120 g at the beginning of the experiment, were fed during 7 months with one of the following diets, containing 72 p. 100 (w/w) carbohydrate: starch, fructose, glucose and sucrose. These diets were about 18% (w/w) in protein content and were conveniently balanced with respect to vitamins and mineral nutrients. After an overnight fast, the animals received by the i.p. way, 30 mn before their killing, one of the following lipogenic precursors: glucose (considered as the control treatment), fructose ethanol or acetate, thus forming 16 experimental groups. In their liver, heart and blood were determined the concentrations of 6 lipid compounds: triglycerides, non-esterified fatty acids, free, esterified and total cholesterol, phospholipide. a. The sucrose diet gave the heaviest animals, with a liver and heart which were the richest in triglyceride content. They had also the highest liver and blood cholesterol, but their blood phospholipid was the lowest. The starch diet also increased, compared to the glucose diet, liver and heart triglycerides and liver cholesterol. As regards the fructose diet, it had the same effects than the sucrose one in elevating liver weight, blood triglycerides and cholesterol; conversely, it lowered liver and chiefly heart triglycerides and increased blood phospholipide. The glucose diet was for almost all parameters the one which displayed the lowest values. b. Relative to the glucose injection, other ones increased liver triglycerides, cholesterol and phospholipids and non-esterified fatty acids of the 3 assayed tissues. We observed that some differences between the effects of two given injections varied according to the previous diet, e.g. the sucrose-fed rats had more liver triglycerides and cholesterol, more heart and blood cholesterol after i.p. fructose than after i.p. glucose, which was not the case for the starch-fed animals. The importance of liver esterification reactions, which are increased with a long-term administration of a fructose-containing diet, is emphasized in the discussion. However dietary fructose could not be able to display its lipogenic effets in the absence of dietary glucose and that is why sucrose is more efficient than glucose in promoting a net lipid synthesis.

Acetates↗
PubMed1975

[In vitro intestinal absorption of sugars in the rat. Changes in concentrations of adenine nucleotides and inorganic phosphate in mucosa].

The absorption of three sugars by the rat gut: glucose, fructose and galactose, was studied in vitro' with a jejunal everted loop technique. The concentrations of the sugars in the serosal medium and in the mucosa cells were measured at given times, as well as the concentrations of ATP, ADP, AMP and inorganic P (Pi) in the mucosa cells. The time course of these concentrations was analyzed with the aid of polynomial regression, using the least squares method.

Adenine Nucleotides↗