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T Barber

Publications and source records attributed to T Barber.

At least 37 records · Page 2Linked to original sources
PubMed1995

Perilipin is located on the surface layer of intracellular lipid droplets in adipocytes.

Immunocytochemistry was used to determine the intracellular location of perilipins in adipocytes and the occurrence of these proteins in tissues involved in triacylglycerol metabolism. Confocal microscopy and 3-dimensional analysis of 3T3-L1 adipocytes showed that perilipin immunofluorescence, present on the surfaces of all sized lipid droplets, appeared unevenly dispersed on the surfaces of many large lipid droplets. Electron microscopy revealed that immunogold staining for perilipin was located directly on the surface layer apposed to and surrounding the core triacylglycerol of intracellular lipid droplets of adipocytes in culture or from white and brown adipose tissue. Freeze-fracture electron microscopy indicated that the hydrophobic face of this surface monolayer contained particles identical in size and distribution to intramembranous particles (IMPs), which are unique structural features of the hydrophobic faces of bilayered membranes. Also, freeze-fracture replicas revealed areas of continuity between the surface layer of lipid droplets and the membrane leaflets of endoplasmic reticulum, suggesting that the droplet monolayer surface is an area of endoplasmic reticulum membrane leaflet modified by its unique content of perilipin. Microperoxisomes, identified by immunostaining for catalase, were found closely associated with lipid droplets, but external to and not in contact with the lipid droplet surface layer. Vimentin, identified by immunofluorescence, was present around the periphery of most lipid droplets in 3T3-L1 cells during early stages of adipocyte development but, in contrast to perilipins, vimentin was not around the periphery of many large lipid droplets in mature cells. Although perilipin was at the surface of lipid droplets in adipocytes of lactating mammary gland, none was found to be associated with the milk lipid droplets in alveolar epithelial cells, nor was the protein found on the surfaces of lipid droplets in hepatocytes. Studies in mammary gland show that perilipin immunostaining will be a valuable tool for the identification of tissue adipocytes severely depleted of their triacylglycerol stores and thus without their characteristic spherical shape. Perilipin's singular location on the surface monolayer of intracellular lipid droplets supports an intimate role for the protein in the triacylglycerol metabolic functions of adipocytes.

3T3 Cells↗
PubMed1994

Microtubules are not required for glucocorticoid receptor mediated gene induction.

Steroid-free glucocorticoid receptors are generally considered to reside in the cytoplasm of cells. After the binding of steroids, the receptors translocate into the nucleus in a manner that has been proposed to involve microtubules. However, some results with inhibitors of microtubule assembly argue to the contrary. In all of these studies, only the whole cell localization of receptors has been examined; the biological activity of these receptors has not been determined. We now report that steroid-induced gene expression is maintained in the absence of intact microtubules. This argues that microtubules are not required for either the nuclear translocation or biological activity of glucocorticoid receptors.

Animals↗
PubMed1994

Protein synthesis in vivo in rats fed on lipid-rich liquid diets.

Changes in tissue composition and protein synthesis ratio were studied in the major tissues of the body in young rats fed on lipid-rich, isonitrogenous purified liquid diets, a convenient method for inducing voluntary overfeeding under controlled nutritional conditions. Overfed rats showed faster growth induced by the energy excess. Analysis of tissue composition (protein, DNA and RNA contents) revealed that growth was due mainly to tissue hyperplasia in which protein and DNA contents increased in parallel. Fractional protein synthesis ratio measured in vivo by the flooding-dose method of phenylalanine showed a marked increase in all tissues. This change could be attributed to an increase in the ribosomal activity for protein synthesis in most tissues. Therefore, our results indicate that addition of a supplementary energy source (as lipids) to a well-balanced diet improves growth and protein synthesis in growing rats.

Animal Nutritional Physiological Phenomena↗
PubMed1994

Improved nitrogen metabolism in rats fed on lipid-rich liquid diets.

N metabolism was studied in young rats fed on lipid-rich, isonitrogenous, purified liquid diets, a convenient and easy technique for inducing voluntary overfeeding of energy and lipids under controlled nutritional conditions. Overfed rats showed a marked N retention at the expense of a reduced production of urea. The capacities of isolated hepatocytes to synthesize urea and glucose from added precursors were greatly diminished. The activities of the urea cycle enzymes and several enzymes involved in the availability of NH3 for this pathway were concomitantly reduced in overfed animals. Therefore, our results showed an improved N metabolism in overfed rats promoted by the overfeeding of lipids that could be due to an enhanced biosynthetic utilization and a reduced catabolism of amino acids. In addition, the versatile and accurate technique for inducing overfeeding in young rats used in the present study could have many advantages for nutritional studies.

Alanine Transaminase↗
PubMed1994

Hepatic amino acid uptake is decreased in lactating rats. In vivo and in vitro studies.

To study the redistribution of amino acids to the mammary gland during lactation we used lactating and virgin rats fed liquid diets. Virgin rats were divided in two groups: one group was fed daily a diet containing the same amount of protein that was consumed the previous day by lactating rats (high protein diet-fed rats), and the other virgin group was fed the normal liquid diet (control). The hepatic availability of amino acids was significantly higher in the lactating rats than in the other two groups, but the uptake and fractional extraction of amino acids by the liver were lower in lactating rats than in the high protein-fed virgin controls. When primary hepatocyte cultures were used, the uptake of 2-amino-[1-14C]isobutyric acid (AIB) and the activity of system A were found to be significantly higher in the hepatocytes from virgin rats fed the high protein diet than in those obtained from the lactating and control virgin groups. No difference was observed between the control virgin rats and the lactating rats. The kinetic of AIB showed that the Vmax/Km ratio was significantly lower in hepatocytes from lactating rats than in those from the high protein diet-fed virgin rats. Addition of prolactin to the incubation medium decreased the uptake of AIB in hepatocytes from both groups of virgin rats. Moreover, uptake of AIB was greater in bromocriptine-treated lactating rats and in lactating rats that had had their pups removed for the preceding 24 h compared with values for the lactating rats.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acids↗
PubMed1992

Nitrogen metabolism in obesity induced by monosodium-L-glutamate in rats.

We have studied the effects of subcutaneous administration of monosodium-L-glutamate (MSG) to neonatal rats on nitrogen metabolism and on general parameters at several intervals after MSG treatment. As MSG-treated rats were hypophagic, all experiments were performed both in control rats pair-fed with the MSG-treated rats and in control rats fed ad libitum. Lee index, total serum lipids and weight of the epididymal fat depots were higher in MSG-treated rats. Body and tissue weights and the amount of protein in several tissues were lower in adult MSG-obese rats than in control rats. Locomotor activity was decreased following MSG administration. Creatinine clearance was diminished by about 50% in rats treated with MSG. Urinary nitrogen and urea excretion were lower, except at four weeks, and serum urea was higher in MSG-obese rats. Considering liver size, urea synthesis by isolated hepatocytes and urea cycle enzyme activities were increased in weanling MSG obese rats and diminished in adult MSG-obese rats when compared with ad libitum controls but were not changed compared with their pair-fed controls. It is concluded that administration of monosodium-L-glutamate shortly after birth induced an increase in urea synthesis in weanling rats that was followed by a reduction in the amount of tissue proteins, suggesting that more amino acids were used for lipid synthesis and urea production in treated rats. The accelerated amino acid degradation slowed down in adult MSG-obese rats which showed an in vitro capacity to synthesise urea similar to that of their pair-fed controls.

Analysis of Variance↗
PubMed1990

Amino acid metabolism and protein synthesis in lactating rats fed on a liquid diet.

1. Amino acid metabolism was studied in control virgin rats, lactating rats and virgin rats protein-pair-fed with the lactating rats (high-protein virgin rats). 2. Urinary excretion of nitrogen and urea was higher in lactating than in control virgin rats, and in high-protein virgin rats it was higher than in lactating rats. 3. The activities of urea-cycle enzymes (units/g) were higher in high-protein virgin than in lactating rats, except for arginase. In lactating rats the activities of carbamoyl-phosphate synthase, ornithine carbamoyltransferase and argininosuccinate synthase were lower than in control virgin rats. When the liver size is considered, the activities in lactating rats were similar to those in high-protein virgin rats, except for arginase. 4. N-Acetylglutamate content was higher in high-protein virgin rats than in the other two groups. 5. The rate of urea synthesis from precursors by isolated hepatocytes was higher in high-protein virgin rats than in the other two groups. 6. The flooding-dose method (L-[4-3H]phenylalanine) for measuring protein synthesis was used. The absolute synthesis rates of mammary gland, liver and small-intestinal mucosa were higher in lactating rats than in the other two groups, and in high-protein virgin rats than in control virgin rats 7. These results show that the increased needs for amino acids during lactation are met by hyperphagia and by a nitrogen-sparing mechanism.

Amino Acids↗
PubMed1989

Effect of a hyperlipidic diet on lipid composition, fluidity, and (Na+-K+)ATPase activity of rat erythrocyte membranes.

Feeding rats a hyperlipidic diet in which animals were offered daily a variety of high-energy food resulted in a significant increase of serum free fatty acids and a decrease of phospholipids with respect to controls. On the contrary, there were no significant differences in erythrocyte membrane total lipid composition between the two groups. Erythrocyte membranes showed a significant decrease in saturated fatty acid content and a significant increase in (n-6) polyunsaturated fatty acid content; (n-3) polyunsaturated fatty acids significantly decreased. Membrane fluidity, investigated by fluorescence polarization of diphenylhexatriene, significantly increased in the erythrocyte membranes of the experimental group. These results seem compatible with the decreased saturated/unsaturated fatty acid ratio. A significant decrease of (Na+-K+)ATPase activity occurred in erythrocyte membranes of the experimental group rats with respect to the controls.

Animals↗
PubMed1989

Lipid composition, fluidity and enzymatic activities of rat liver plasma and mitochondrial membranes in dietary obese rats.

Total lipids and fatty acid composition were determined in liver plasma and mitochondrial membranes from control and dietary obese rats after 4 weeks of the experimental period. The lipid composition of liver plasma and mitochondrial membranes showed an increase of triacylglycerols in obese rats. The liver plasma membranes showed a decrease of saturated/unsaturated fatty acid ratio and an increase of (n-6) polyunsaturated fatty acids, whereas the (n-3) polyunsaturated acids were decreased. Contrary to what occurs with plasma membranes, few modifications were observed in mitochondrial membranes. Changes of the fatty acid composition of the phospholipid bilayer are of potentially great importance in structural and functional parameters of membrane. Fluidity of liver plasma membranes of dietary obese rats was highly increased, while the mitochondrial ones remained unchanged. These results can be well explained by the decreased saturated/unsaturated fatty acid ratio. A significant decrease of (Na+-K+) ATPase activity (a membrane bound enzyme) was found in plasma membranes of dietary obese rats. Mitochondrial enzymatic activities and oxidative phosphorylation showed few changes except a small, but significant decrease of state 3 respiratory rate. In this study we also determined the fatty acid composition of all the foods offered to animals and their daily intakes in order to discuss their possible influence on changes in structural and functional membrane parameters.

Animals↗
PubMed1988

An enzyme immunoassay for the quantitation of rat liver carbamoyl-phosphate synthetase I.

An indirect, competitive enzyme-linked immunosorbent assay for the quantitation of carbamoyl-phosphate synthetase I (ammonia) in rat liver has been developed. Homogenization of the liver in 1% sodium deoxycholate is used for complete solubilization of the enzyme. The detergent does not interfere with the method if diluted to a concentration of 0.01% or lower. The assay is applied to determine the amount of enzyme in control rats and in rats fed "cafeteria" or high-protein diets. Changes in the amount of carbamoyl-phosphate synthetase I (ammonia) paralleled changes in enzymatic activity.

Animals↗
PubMed1987

Studies on the role of insulin in N metabolism changes in cafeteria-fed rats.

The present work studies the serum insulin response to cafeteria feeding and the possible role of this hormone in the marked N retention induced by cafeteria feeding. Rats fed a cafeteria diet for periods of 10, 20 and 30 days showed a significant and progressive increase in serum insulin levels. In order to elucidate the possible intervention of this hormone in the marked N retention induced by cafeteria feeding we studied the changes in N metabolism in non-diabetic and streptozotocin-diabetic rats fed a cafeteria diet for 10 days. The amount of N ingested was the same in all experimental groups. Our results suggest that insulin is not absolutely indispensible in the N retention exhibited by cafeteria-fed rats, inasmuch as cafeteria feeding also promotes a decrease in N excretion in streptozotocin-diabetic rats. This is in accordance with the reduction in urea excretion and the activities (U/g liver) of the ureogenic enzymes in streptozotocin-diabetic rats fed a cafeteria diet.

Animals↗
PubMed1985

Decreased urea synthesis in cafeteria-diet-induced obesity in the rat.

Feeding rats with a cafeteria diet resulted in increases in total body weight and in epididymal-adipose-tissue weight. Those rats excreted significantly less N than did controls. The amount of N ingested by cafeteria-diet-fed rats was kept equal to that of controls. This decrease in N excretion is explained by a decrease in urinary excretion of urea. This may be due to the following facts. The rate of synthesis of urea from precursors by isolated hepatocytes from cafeteria-diet-fed rats was lower than in controls. In cafeteria-diet-fed rats the activities of all the enzymes of the urea cycle are decreased. The major percentage decreases are those of carbamoylphosphate synthetase (EC 6.3.4.16) and of argininosuccinate synthetase (EC 6.3.4.5), the enzymes probably involved in the regulation of the overall rate of the cycle. When rats are switched to normal chow diet, the enzyme activities return to normal values. The uptake of amino acids by liver of cafeteria-diet-fed rats is lower than in controls. These results contrast with those obtained previously by using other models of obesity in rat (i.e. genetic or hypothalamic), in which N excretion was increased.

Amino Acids↗
PubMed1982

Variability in urinary mercury excretion.

This report investigates quantitatively the extent to which 24-hour urinary mercury excretion can be predicted from the concentration of mercury in spot samples. The correlation between these quantities can be significantly improved by the application of a correction for urinary dilution and by restricting spot samples to the day's first excretion. If these precautions are taken, we find that for 68% of the samples the 24-hour excretion, Q, is in the range of 0.75 C' less than Q less than 1.31 C', where C' is the mercury concentration of a day's first excretion. The width of this range appears to be primarily determined by variations in a person's daily urinary volume and by the frequent occurrence of diurnal cycles.

Environmental Exposure↗
PubMed1976

Morula forms of E rosettes: distribution, specific inhibition, and enhancement.

Rosetts-forming cells (RFC) with more than 10 sheep red blood cells (SRBC's) per cell defined as morula-forming cells (MFC) were studied in human lymphoid organs and in the peripheral blood (PBL). MFC are most common in the thymus (more than 50%) and the frequency is much less in peripheral lymphoid tissues and in the blood (29.8% in those less than 5 years old; 14.9% in adults). There is very slight decrease in total RFC in PBL over age 5, but the changes in MFC are quite striking. Treatment with antihuman thymus antiserum (ATS) and with anti-human lymphoblastic antiserum (ATL) resulted in significant inhibition of total rosette and morula rosette formation. No inhibition was seen using normal brain antiserum, anti-IgM antiserum, or anti-L chain antiserum. In two patients with chronic lymphocytic leukemia and in some cases of thymic deficiency the ratio of MFC to total RFC was unusually high. After culture with phytohemagglutinin (PHA) and conversion to blast forms or after surface peturbation by papain or freeze-thaw treatment, MFC numbers were increased.

Humans↗