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Biomedical subjects

K Kaya

Publications and source records attributed to K Kaya.

At least 73 records · Page 4Linked to original sources

Different incorporation rates of arachidonic acid into alkenylacyl-, alkylacyl- and diacylphosphatidylethanolamine of rat erythrocytes.

Rat erythrocyte phosphatidylethanolamine (PE) consists of 60% alkenylacyl, 5% alkylacyl and 35% diacyl types. The fatty acid at the 2-position of these types is mainly composed of arachidonic acid. When intact rat erythrocytes were incubated with exogenous arachidonic acid, about 90% of the arachidonic acid incorporated into the PE fraction was found in the 2-position of the diacyl type. The rates of incorporation of arachidonic acid into alkenylacyl-, alkylacyl- and diacylPE were 78, 134 and 1360 pmol/h per mumol of the corresponding PE, respectively. The substrate specificities of endogenous phospholipase A2 and acyl-CoA:lysophospholipid acyltransferase were observed. DiacylPE was hydrolysed rapidly by endogenous phospholipase A2, while alkenylacyl- and alkylacylPE were poor substrates for the enzyme. The selective transfer of arachidonic acid into the 2-position of 1-acyl-lysoPE was observed. 1-Alkenyl- and 1-alkyl-lysoPE were also poor substrates for acyl-CoA:lysophospholipid acyltransferase. The acyltransferase activities with the lysoPE analogues were higher than the phospholipase A2 activities with PE analogues. These results suggest that the different incorporation rates of arachidonic acid into alkenylacyl-, alkylacyl- and diacylPE are based on the substrate specificity of endogenous phospholipase A2.

Animals↗

Effects of nitrate and nitrite, chemical intermediates of inhaled nitrogen dioxide, on membrane components of red blood cells of rats.

Rat blood was incubated at 37 degrees C for 60 min with either NaNO3 or NaNO2 to examine the relationship between the decrease in the hexose content and Ca2+,Mg2+-ATPase activity of red cell membranes, and NO3- and NO2-. The hexose content decreased depending on the NaNO2 concentration up to 100 microM reaching 76% (p less than 0.05) of the control value. NaNO3 had little effect on the hexose content. On the other hand, the Ca2+,Mg2+-ATPase activity decreased depending on the NaNO3 concentration up to 200 microM, where the activity reached 75% (p less than 0.01) of the control value. The effect of NaNO2 on this activity was smaller than that of NaNO3. The sialic acid content and the Na+,K+-ATPase activity did not show significant alterations by incubation with NaNO2 and NaNO3 at below 100 microM. To examine the in vivo effects of NO2- and NO3-, 50 mM NaNO3 was intravenously injected into rats five times at hourly intervals (dose: 1.0 ml/kg body weight), and blood was collected 1 hr after the last injection. The activities of Ca2+,Mg2+- and Na+,K+-ATPases of red cell membranes were decreased to 68% (p less than 0.05) and 80% of the control value, respectively. Reduction by injection of 50 mM NaNO2 was smaller than that by 50 mM NaNO3. The results show that the hexose content and the Ca2+,Mg2+-ATPase activity of red cell membranes were decreased by NO-x that increased in the blood during short-term exposure of rats to NO2.

Animals↗

On the formation of alpha-hydroxy fatty acids. Evidence for a direct hydroxylation of nonhydroxy fatty acid-containing sphingolipids.

When Tetrahymena pyriformis, strain NT-1, was warmed from a growth temperature of 15 degrees C to 39 degrees C, there was a rapid conversion of its sphingolipids from the nonhydroxy fatty acid-containing ceramide aminoethylphosphonate (NCAEP) predominant at low temperature to the alpha-hydroxy fatty acid-containing analog (HCAEP). The fatty acid composition of the freshly formed HCAEP strongly resembled that of the original NCAEP pool. An extensive utilization of NCAEP long chain bases for HCAEP formation was shown by prelabeling the NCAEP bases with [14C]serine. The specific radioactivities of both fatty acids and long chain bases of the two sphingolipid classes of cells prelabeled with [3H]palmitic acid at 15 degrees C and then warmed to 39 degrees C were compatible with a direct hydroxylation of the intact ceramide aminoethylphosphonate or free ceramide formed from it. Exogenously added alpha-hydroxypalmitic acid was not incorporated into the sphingolipids intact, but there was an active alpha-oxidation of alpha-hydroxy fatty acids, yielding nonhydroxy fatty acids one carbon atom shorter in length. In vitro experiments failed to demonstrate a direct hydroxylation of NCAEP, and nonhydroxy fatty acid-containing free ceramides were hydroxylated only sparingly. This inefficient hydroxylation is attributed to the rapid enzymatic hydrolysis of these substrates in vitro and to a limited availability of the added ceramides to the hydroxylating enzymes. The weight of the evidence from this study strongly favors the alpha-hydroxylation of fatty acids only when they are bound as elements of sphingolipids.

Animals↗

Composition and metabolism of fatty acids in phospholipids of density-separated red cells of rats.

Fatty acid compositions of phosphatidylcholine (PC) and phosphatidylethanolamine (PE) and the rates of fatty acid esterification to these phospholipids (PL) were measured in intact rat red cell populations of different ages separated by density gradient centrifugation in order to clarify changes in membrane lipids of red blood cells during in vivo aging. Fatty acid compositions of PC and PE altered progressively as red cells became denser. Changes in unsaturated fatty acids occurred predominantly at the 2-position of PC and PE and those in saturated fatty acids at both positions. The esterification rates of 5 major fatty acids decreased as red cells became denser and those of oleic acid, linoleic acid and arachidonic acid to both PC and PE of fraction I cells (oldest cells) were 37-51% those of fraction IV cells (youngest cells). Reduction in the rates of fatty acid esterification appeared to occur in the course of red cell maturation because reticulocyte-enriched cell fractions showed 4.5-14.5 times higher rates of linoleic acid and arachidonic acid esterifications to PC and PE.

Animals↗

In vivo effects of nitrogen dioxide on membrane constituents in lung and liver of rats.

When rats were exposed to 10 ppm NO2 for 7 days, the succinate-cytochrome c reductase activity, the rate-limiting step of mitochondrial succinoxidase, of the liver decreased progressively, reaching 77% (P less than 0.01) of the control level by the fifth day. By contrast, this activity was gradually increased by a 10-day exposure to 4 ppm NO2 and reached 1.14-fold that of the control level at the seventh day. A reduction in the components of electron-transport systems in liver microsomes was found during exposure to NO2 at both concentrations. This reduction was preferential for components of drug-metabolizing systems. The NADPH-cytochrome c reductase activity and the cytochrome P-450 content were decreased to 82% (P less than 0.05) and 76% (P less than 0.05), respectively, at the fifth day of exposure to 10 ppm NO2. Exposure to 4 ppm NO2 also caused a significant decrease in the NADPH-cytochrome c reductase and the cytochrome P-450 content, which were lowered to 84% (P less than 0.05) of the control level at the fourth and seventh days, respectively. Reduction in these components appears to occur even at the concentration of 0.4 ppm at the seventh day. Alterations in the components of mitochondria and microsomes of the lung were considerably different from those of the liver. The protein content of the lung increased to 1.18 (P less than 0.05)- and 1.14 (P less than 0.05)-fold that of the control during exposure to 10 and 4 ppm NO2, respectively. The succinate-cytochrome c reductase activity showed a reduced value, which was 73% (P less than 0.01) of the control level, 1 day after exposure to 10 ppm NO2, Subsequently, the activity increased to 1.14-fold that of the control during exposure to 10 (at the fifth day) and 4 ppm (at the fourth day) NO2. Components of microsomal electron-transport systems also showed a slightly elevated value during exposure to 4 ppm NO2. Seven days after exposure, the cytochrome P-450 content was decreased to 61% (P less than 0.01) of the control level, while other components were retained at control levels. A significant reduction in the cytochrome P-450 content was also observed at the exposure concentration of 1.2 ppm, but not of 0.4 ppm, at the seventh day.

Animals↗

Effects of nitrogen dioxide on red blood cells of rats: alterations of cell membrane components and populational changes of red blood cells during in vivo exposure to NO2.

Male Wistar rats were exposed to 4 ppm NO2 for 10 days in order to examine the relationship between the changes in components of red cell membranes and alterations of erythrocyte population. Na+, K+-ATPase activity of red blood cell membranes of exposed animals showed a significantly higher value than that of the control at the first and fourth days of exposure and then decreased to under the control value at the seventh day. In order to examine changes in erythrocyte population, red blood cells were fractionated into four fractions according to their density using Dextran density centrifugation. The alteration of the percentage of lowest-density cells (fraction IV) of exposed animals was completely consistent with that of Na+,K+-ATPase activity in addition to that of the sialic acid content as described in a previous report (K. Kaya, T. Miura, and K. Kubota (1980). Environ. Res. 23, 397-409.). The percentage of fraction IV was 1.43- (P less than 0.05) and 1.68-fold (P less than 0.01) those of the control at the first and fourth days of exposure, respectively, and then decreased to under the control value at the seventh day. This decrease accompanied increases in the percentages of higher-density cells (fractions I and II). Examination of subfractions of red blood cells showed that Na+,K+-ATPase activity and the sialic acid content of three fractions with lower densities have higher values in exposed animals than in the control 1 day after exposure to NO2. Based on these results, it is concluded that increases in Na+,K+-ATPase activity and the sialic acid content occurring 1 day after exposure to 4 ppm NO2 were caused by elevated levels of these components in three fractions with lower densities as well as by an increase in the percentage of lowest-density cells in the erythrocyte population. It was also suggested that NO2 inhalation accelerated aging of erythrocytes with respect to density. The change in Ca2+,Mg2+-ATPase activity, in addition to that in the hexose content as described in a previous report (Kaya et al., 1980), was different from those in the sialic acid content and Na+,K+-ATPase activity. Ca2+,Mg2+-ATPase activity and the hexose content of exposed animals showed slightly reduced values 1 day after exposure to NO2. In all subfractions of red blood cells these values were slightly lower in exposed animals than in the controls. Therefore, reduction in Ca2+,Mg2+-ATPase activity and the hexose content is not due to changes in erythrocyte population.

Animals↗

Temperature-induced changes in the hydroxy and non-hydroxy fatty acid-containing sphingolipids abundant in the surface membrane of Tetrahymena pyriformis NT-1.

Sphingolipids make up 30 to 40 mole % of the phospholipids found in the surface membrane of Tetrahymena pyriformis NT-1. We have identified the two major classes as non-hydroxy fatty acid-containing ceramide-2-aminoethylphosphonate (NCAEP) and alpha-hydroxy fatty acid-containing ceramide-2-aminoethylphosphonate (HCAEP). Both classes were well represented in cells grown at 39 degrees C. At this temperature their principal long chain bases were n-hexadeca-4-sphingenine and n-nonadeca-4-sphingenine. The major fatty acid of NCAEP from 39 degrees C-grown cells was palmitic acid and that of HCAEP was alpha-hydroxypalmitic acid. Cells grown at 15 degrees C contained NCAEP, but only traces of HCAEP. By analyzing the incorporation of [1-14C]palmitic acid into cells growing isothermally or shifted from 15 degrees C to 39 degrees C, we obtained evidence favoring a direct conversion of NCAEP to HCAEP. This conversion was blocked in cells grown at 15 degrees C, causing an accumulation of NCAEP. Tetrahymena is a useful model system for studying the poorly understood alpha-hydroxylation process that is of critical importance in myelination of animal nervous tissues.

Animals↗

Enkephalin degrading enzymes in cerebrospinal fluid.

Enkephalins were rapidly degraded by specific enzyme systems in vivo. In cerebrospinal fluid (CSF), however, it has been undefined whether these enzyme systems existed. Our experiments showed enkephalins were hydrolyzed by the enzymatic activity in both CSF of human and monkey. The results by the thin layer chromatography and the high performance liquid chromatography revealed the reaction products of CSF and enkephalin were tyrosine, tyrosyl-glycine and tyrosyl-glycyl-glycine. Therefore, the enzymes in CSF were considered to be an aminopeptidase, a dipeptidyl aminopeptidase and a dipeptidyl carboxypeptidase. Our results suggest that in the assay of enkephalin in CSF, the effects of these enzymes should be considered.

Cerebrospinal Fluid↗

Selective changes in fatty acid composition of phosphatidylserine in rat erythrocyte membrane induced by nitrate.

The relationship between nitrate which is formed from inhaled nitrogen dioxide, a common air pollutant, and changes in fatty acid metabolism of phosphatidylserine in rat erythrocytes has been examined. When erythrocytes were incubated at 37 degrees C for 60 min with fatty acid, the incorporation rate of [1-14C]arachidonic acid and [9,10-3H]palmitic acid into phosphatidylserine was 15% (80 pmol/h per mumol lipid phosphorus) and 20% (12 pmol/h per mumol lipid phosphorus) of those into phosphatidylethanolamine, respectively. By the addition of 1.0 mM sodium nitrate or 0.5 microM ionophore A23187 to the incubation mixture, the rate of incorporation of both arachidonic acid and palmitic acid into phosphatidylethanolamine was stimulated 1.45-fold. On the other hand, the incorporation of palmitic acid into phosphatidylserine was little affected, while that of arachidonic acid was stimulated 1.35-fold. An increase in arachidonic acid of phosphatidylserine was also found by the addition of nitrate or ionophore A23187. This increase was dependent on the concentration of extracellular calcium and observed by the addition of other chaotropic anions in the order SCN- greater than ClO4- greater than NO3-. It seems likely, therefore, that nitrate causes changes in erythrocyte membranes to facilitate calcium uptake. Increasing the concentration of intracellular calcium may cause stimulation of acyl-CoA:lysophospholipid acyltransferase and/or endogenous phospholipase A2.

Animals↗

Short term improvement in glycemic control utilizing continuous subcutaneous insulin infusion: the effect on 24-hour integrated concentrations of counterregulatory hormones and plasma lipids in insulin-dependent diabetes mellitus.

Eight patients with established insulin-dependent diabetes mellitus were studied before and 2 weeks after the initiation of pumped continuous sc insulin infusion in order to investigate the effect of short term improvement of glycemic control on hormonal and lipid levels. Glycemic control was improved in all patients. Using a constant blood withdrawal pump, accurate 24-h average concentrations, denoted integrated concentrations, were obtained. The mean 24-h integrated concentrations of GH, cortisol, norepinephrine, and epinephrine did not change significantly. The mean fasting triglyceride concentration dropped from 119.1 to 83.4 mg/dl (P less than 0.05). The mean 24-h integrated concentration of plasma triglycerides fell from 132.1 to 101.5 mg/dl (P less than 0.02). Both mean fasting and mean 24-h integrated concentrations of plasma cholesterol were lower after improved control. Short term improvement in glycemic control was associated with a reduction in plasma lipid concentrations, but failed to alter mean 24-h integrated concentrations of the measured counterregulatory hormones.

Adolescent↗