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Biomedical subjects

R Curi

Publications and source records attributed to R Curi.

At least 109 records · Page 6Linked to original sources

Changes in the activities of antioxidant enzymes of the lymphoid organs of 21-day pregnant rats due to administration of fish oil by gavage.

1. The effect of fish oil administration by gavage (0.4% body weight) on activities of superoxide dismutase (SOD), catalase and glutathione peroxidase (GSH-Px) and on content of thiobarbituric acid reactive substances (TBARs) of the lymphoid organs [thymus, spleen and mesenteric lymph nodes (MLN)] and liver was investigated in 21-day pregnant rats. The results were compared with those obtained by administration of soybean oil, cocoa butter and coconut oil. 2. Oil administration did not have any significant effect on antioxidant enzyme activities of the liver, whereas marked changes were found in the lymphoid organs. The MLN presented the most pronounced changes: SOD and catalase activities were increased by the four oils; GSH-Px activity was raised by soybean and fish oils; coconut oil reduced the activity of the three antioxidant enzymes in this organ. 3. Fish oil given by gavage does affect the antioxidant capacity of the lymphoid organs; however, similar effect was also observed for cocoa butter and soybean oil. These changes in the antioxidant enzyme activities were able to prevent the lipid peroxidation process in the lymphoid organs.

Animals↗

Acetate and propionate potentiate the antiproliferative effect of butyrate on RBL-2H3 growth.

1. The effect of acetate, propionate, and butyrate separately and combined on RBL-2H3 (a rat basophilic leukemic cell type) proliferation during 24, 48, and 72 hr was examined. Also, the effect of a mixture of the three volatile fatty acids on proliferation of HeLa-155 (a human adenocarcinoma), C57 B1/6J (a mouse melanoma), and MCF-7 (human breast tumor) during 8 days was investigated. 2. Acetate and propionate per se did not present any effect on RBL-2H3 growth during 72 hr, however, when acetate and propionate were added together a significant inhibition of this cell growth was found; 18% for 48 and 37% for 72 hr. The addition of butyrate to the culture medium caused a 75% decrease in the rate of this cell growth either after 48 and 72 hr. This effect of butyrate was pronounced by acetate (86% and 90% for 48 and 72 hr, respectively), propionate (87% for 48 and 93% for 72 hr), and acetate and propionate together (76% for 48 and 92% for 72 hr). 3. Daily addition of a mixture of the short-chain fatty acids (10 mM acetate, 2 mM propionate and 1.5 mM butyrate) markedly decreased the number of cells after 8 days: 58% for RBL-2H3, 42% for HeLa-155, 91% for C57 B1/6J and 55% for MCF-7. 4. These results support the proposition that a fiber-rich diet that leads to great production of butyrate but also of propionate and acetate would be more effective to prevent the occurrence of colorectal cancer than the administration of this short-chain fatty acid given alone.

Acetates↗

Metabolic fate and effects of saturated and unsaturated fatty acids in Hep2 human larynx tumor cells.

Previous studies have reported the presence of carnitine palmitoyltransferase I and II in tumor cells and the inhibitory effects of fatty acids on cell proliferation. The present work considered the metabolic fate of [14C] or [3H]-labeled fatty acids and their effects on cellular metabolism in Hep2 human larynx tumor cells. The rate of uptake of acetate was 45% of that of myristate, palmitate, oleate, linoleate and arachidonate. However, acetate was rapidly metabolized within the cell as seen by its low rate of accumulation as non-esterified fatty acid, < 5% of that of the other fatty acids. The incorporation of fatty acids into neutral lipid fractions showed palmitate and oleate primarily entered the phospholipid fraction, while linoleate and arachidonate entered equally the phospholipid and triacylglycerol fractions. Palmitate and oleate were oxidized to 14CO2 at higher rates than linoleate and arachidonate, with arachidonate being the least oxidized of the unsaturated fatty acids. Acetate was oxidized at 10-30 fold higher rates than the other fatty acids. Palmitate, oleate, linoleate and arachidonate all had significant inhibitory effects on the rate of glucose utilization by Hep2 cells, ranging from 25-38% inhibition and were found to inhibit cell proliferation by 17-73%. These findings suggest that certain fatty acids not only play a structural role in cellular metabolism, but may also have a potential regulatory role in the glycolytic pathway of Hep2 cells.

Fatty Acids↗

Evidence for the transfer in culture of [14C]-labelled fatty acids from macrophages to lymphocytes.

[14C]-labelled palmitic acid (PA), oleic acid (OA), linoleic (LA) and arachidonic (AA) acids were transferred from macrophages (M phi) to lymphocytes (LY) when equal numbers of the two cell types were co-cultured. The relative degree and amounts of the fatty acids transferred from M phi to LY are as follow: AA (368.57 +/- 21.62) = OA (274.52 +/- 15.41) > LA (42.11 +/- 8.31) = PA (36.53 +/- 2.45). The transfer units are nmol/10(10) M phi/10(10) LY and the values are mean +/- SEM for 7 experiments. The [14C]-radioactivity transferred was mainly directed to the phospholipid fraction of the lymphocytes (85% by PA, 86% by LA, 83% by OA and 79% by AA). In the same order as above, phosphatidylcholine was the phospholipid moiety most heavily labelled (82% by PA, 71% by LA, 66% by OA and 47% by AA). The amount of [14C]-radioactivity transferred to stimulated lymphocytes of thioglycollate treated animals remained unchanged for LA, PA and AA but reduced for OA (71%). The significance of these observations for the immune functions of the cells and resolution of the question of whether some of the [14C]-isotope transfer involves a component of exchange or is unequivocally net fatty acid mass transfer are still being investigated.

Animals↗

Peroxidase activity may play a role in the cytotoxic effect of indole acetic acid.

Peroxidase activity in neutrophils is higher than in thioglycollate macrophages, while in lymphocytes this enzyme activity is very low. Indole-3-acetic acid is oxidized by peroxidase and the role of this enzyme in the cytotoxic effect of the compound was evaluated by measuring oxygen consumption, light emission and cell death in neutrophils, macrophages and lymphocytes. The increase in light emission, oxygen consumption and rate of cell death in cells cultured in the presence of indole-3-acetic acid presented a direct correlation with the peroxidase activity of the cells as follows: neutrophils > thioglycollate macrophages > resident macrophages > lymphocytes. Indeed, in lymphocytes that possess very low peroxidase activity, indole-3-acetic acid did not result in an increase in light emission or oxygen consumption and it was not cytotoxic.

Animals↗

Glutamine utilization by rat neutrophils: presence of phosphate-dependent glutaminase.

The capacity of rat neutrophils to utilize glutamine was investigated by 1) determination of oxygen consumption in the presence of glucose or glutamine, 2) measurement of maximal activity of phosphate-dependent glutaminase, 3) Northern blot, Western blot, and immunocytochemical detection of glutaminase, and 4) measurement of glutamine utilization and also production of ammonia, glutamate, aspartate, alanine, and lactate and decarboxylation of [U-14C]glutamine in cells incubated for 1 h. The rate of respiration by isolated neutrophils in the absence of added substrate was 5.0 nmol x min(-1) x 10(7) cells(-1). Maximal activity of phosphate-dependent glutaminase was 56 nmol x min(-1) x mg protein(-1) in freshly obtained neutrophils; the Michaelis-Menten constant was 3.5 mM for glutamine. This enzyme activity was inhibited by 2 mM glutamate, 2 mM oxoglutarate, and 2 mM NH4Cl. The presence of glutaminase protein (65 kDa) was confirmed by Western blot and immunocytochemical detection and the presence of the mRNA (6.0 kb) by Northern blot analysis. Glutamine was utilized by neutrophils incubated for 1 h at a rate of 12.8 nmol x min(-1) x mg protein(-1) when the amino acid was added to the medium at 2 mM, which is three to four times higher than the physiological concentration. In the presence of 0.5 mM glutamine, the amino acid was utilized at a rate of 2.9 nmol x min(-1) x mg protein(-1). The addition of 0.5 mM glutamate to the incubation medium caused a marked reduction (by 70%) in glutamine utilization by neutrophils. Glucose was utilized at 7.7 nmol x min(-1) x mg protein(-1) when cells were incubated in 5 mM glucose. The conversion of [U-14C]glutamine to 14CO2 was very low: <1% was totally oxidized. The formation of ammonia was approximately 27% of glutamine utilization, and the conversion of glutamine to glutamate, aspartate, alanine, and lactate accounted for approximately 84.6% of the total amino acid utilized by neutrophils. In this study, evidence is presented that, in addition to lymphocytes and macrophages, neutrophils also utilize glutamine.

Animals↗

Effect of protein malnutrition on the glycolytic and glutaminolytic enzyme activity of rat thymus and mesenteric lymph nodes.

The activity of important glycolytic enzymes (hexokinase, phosphofructokinase, aldolase, phosphohexoseisomerase, pyruvate kinase and lactate dehydrogenase) and glutaminolytic enzymes (phosphate-dependent glutaminase) was determined in the thymus and mesenteric lymph nodes of Wistar rats submitted to protein malnutrition (6% protein in the diet rather than 20%) from conception to 12 weeks after birth. The wet weight (g) of the thymus and mesenteric lymph nodes decreased due to protein malnutrition by 87% (from 0.30 +/- 0.05 to 0.04 +/- 0.01) and 75% (0.40 +/- 0.04 to 0.10 +/- 0.02), respectively. The protein content was reduced only in the thymus from 102.3 +/- 4.4 (control rats) to 72.6 +/- 6.6 (malnourished rats). The glycolytic enzymes were not affected by protein malnutrition, but the glutaminase activity of the thymus and lymph nodes was reduced by half in protein-malnourished rats as compared to controls. This fact may lead to a decrease in the cellularity of the organ and thus in its size, weight and protein content.

Animals↗

Wheat bran- but not oat bran-enriched diets increase the mucosal height of the cecum and colon of newly weaned and aged rats.

The effect of diets enriched with oat or wheat bran (prepared by the addition of 300 g of each fiber to 1000 g of the regular diet), given for 8 weeks, on the mucosal height of the colon and cecum was investigated. Newly weaned (21 days old) and aged (12 months old) male Wistar rats were used in this study. As compared to controls, diets enriched with wheat bran provoked a significant increase in the mucosal height, whereas oat bran did not cause any effect. In newly weaned rats (21 days old), wheat bran increased the mucosal height (microns) in the cecum by 20% (mean +/- SEM for 8 rats; 169.1 +/- 5.2 and 202.9 +/- 8.0 for control and wheat bran, respectively) and in the colon (218.8 +/- 7.2 and 264.5 +/- 18.8 for control and wheat bran, respectively). A similar effect was observed in aged rats (12 months old), with an increase of 15% in the mucosal height (microns) of the cecum (mean +/- SEM of 8 rats; 193.2 +/- 8.6 and 223.7 +/- 8.3 for control and wheat bran, respectively) and of 17% in the colon (300.4 +/- 9.2 and 352.2 +/- 15.9 for control and wheat bran, respectively).

Aging↗

Supplementation of aspartate, asparagine and carnitine in the diet causes marked changes in the ultrastructure of soleus muscle.

The effects of the supplementation of aspartic acid and asparagine (45 mg.kg1 body weight of each), and carnitine (90 mg.kg-1 body weight) during one week on the ultrastructure of soleus muscle from swimming-trained (five weeks) and sedentary rats were examined. In trained rats, the amino acids supplementation was performed during the last week of the exercise training only. Supplementation of these amino acids in the diet either in sedentary and trained rats caused myofibrillar and mitochondrial disorganization and dissolution. Focal degeneration of myofibrils and Z-line streaming and disruption, as well as internalization of nuclei were observed. The size of mitochondria increased and some of them presented severe swelling, with decreased electron-density of the matrix and disruption of internal and external membranes. The changes in the soleus muscle ultrastructure described do suggest functional disorders. This observation is particularly important for the amino acid intakers and deserves to be further investigated.

Animals↗

The effect of insulin on macrophage metabolism and function.

This study examined the effect of insulin on rat macrophage metabolism and function. The following parameters were studied: cell migration in response to thioglycollate and BCG stimuli, macrophage phagocytic capacity, H2O2 production, glucose and glutamine metabolism as indicated by the measurement of enzyme activities, the utilization of metabolites and production and oxidation of substrates. The results indicate that insulin: (1) did not affect cell migration in response to thioglycollate and BCG; (2) enhanced the phagocytic capacity of macrophages and the production of H2O2 by macrophages; (3) increased the metabolism of glucose and reduced that of glutaminase.

Alloxan↗

Metabolism of glucose and glutamine in lymphocytes from Graves' hyperthyroid patients: influence of methimazole treatment.

Several studies have shown that thyroid hormones are able to influence selected immune responses such as cell mediated immunity, differentiation of B lymphocytes and the activity of NK cells. These hormones can also regulate the metabolism of glucose and glutamine in rat macrophages and their effects seem to occur mainly through the Krebs cycle. Alterations in the hexokinase, citrate synthase, glucose-6-phosphate dehydrogenase and glutaminase activities in lymphocytes from patients with Graves' disease, either untreated or on methimazole (MMI) therapy were investigated. Experiments were also done in vitro to determine the activities of these enzymes in normal lymphocytes cultured for 24 h in the presence of MMI T3 and T4 using concentrations close to the physiological. Changes in the conversion of [U-14C]-glucose and [U-14C]-glutamine to 14CO2 as caused by the addition of MMI, T3 or T4 to the culture medium were also evaluated. The results indicate that high levels of thyroid hormones might stimulate the metabolism of glucose and glutamine for a short period of time but, if the stimulus is maintained, the utilization of glutamine by lymphocytes is then suppressed. Moreover, MMI does affect lymphocyte metabolism but the significance of this finding for its immunosuppressive effect remains to be examined.

Adult↗

Insulin treatment can abolish changes in glucose and glutamine metabolism of lymphocytes and macrophages caused by the implantation of the Walker 256 tumour.

Activation of lymphocytes and macrophages by the implantation of tumour cells (10(7) cells per rat) into the left flank of rats increased the conversion of glucose to lactate and of glutamine to glutamate and aspartate and the decarboxylation of [U-14C]-glucose and [U-14C]-glutamine in incubated cells. In addition, the amount of GLUT1 was increased in macrophages. The effect of insulin treatment on glucose and glutamine metabolism of lymphocytes and macrophages activated by Walker 256 tumour implantation was also examined. For this purpose, insulin was injected subcutaneously (4 U/100 g b.w. daily) after the fourth day of tumour implantation and the rats were killed 10 days afterwards. Insulin treatment fully reverted the changes due to tumour implantation in the metabolism of glucose and glutamine in lymphocytes and of glucose in macrophages.

Animals↗

Glutamine metabolism is changed in lymphocytes from aged rats.

Changes in the protein content, maximal activity, and Km of phosphate-dependent glutaminase were measured in the lymphoid organs (thymus, spleen, and mesenteric lymph nodes) from just-weaned, mature (3 months), and aged rats (15 months). Also, [U-14C] glutamine transport and decarboxylation and the production of glutamate and aspartate from 2 and 20 mM glutamine were measured in incubated mesenteric lymph node lymphocytes. The ageing process induced a reduction in the protein content of the thymus and spleen, as well as the phosphate-dependent glutaminase activity in the thymus and isolated lymphocytes. The Km of phosphate-dependent glutaminase, however, was not affected by the process. Ageing reduced [U-14C] glutamine decarboxylation and increased glutamate and aspartate production in incubated lymphocytes. The results indicate that the ageing process does modify several aspects of glutamine metabolism in lymphocytes: reduces maximal glutaminase activity and [U-14C] glutamine decarboxylation and increases the Km for [U-14C] glutamine uptake and the production of glutamate and aspartate.

Aging↗