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Biomedical subjects

V Fitzgerald

Publications and source records attributed to V Fitzgerald.

17 recordsLinked to original sources
PubMed1991

Acylation of alkyllysophospholipids by Fischer sarcoma microsomes.

Acylation of alkyllysophospholipids in most cells occurs by: (a) CoA-independent transacylation, (b) CoA-dependent transacylation, and (c) acyl-CoA-dependent acylation. Using a recently developed high-performance liquid chromatography method, we have investigated the factors that influence the molecular species composition of the acylated products formed via these pathways with 1-hexadecyl-2-lyso-sn-glycero-3-phosphocholine (alkyllyso-GPC) or 1-hexadecyl-2-lyso-sn-glycero-3-phospho-ethanolamine (alkyllyso-GPE) as substrates for the enzymes in Fischer R-3259 sarcoma microsomes. We found that short incubation times and low substrate concentrations favored the formation of polyunsaturated molecular species, i.e., 16:0-22:6, 16:0-22:5 (n - 3), and 16:0-20:4. Also, in agreement with results from other systems, CoA-independent transacylation produced a high percentage of polyunsaturated molecular species; acyl-CoA-dependent acylations generated the least polyunsaturated molecular species and CoA-dependent transacylation gave intermediate values. Furthermore, no substrate selectivity occurred with respect to alkyl chain lengths of alkyllyso-GPE; similar molecular species composition was obtained with either hexadecyllyso-GPE or octadecyllyso-GPE as substrates. Responses to N-ethylmaleimide inhibition and heat inactivation as well as pH optima suggest the same enzyme catalyzes the CoA-independent transacylation of both alkyllyso-GPC and alkyllyso-GPE.

Acylation↗
PubMed1990

Regulation of the synthesis of platelet-activating factor and its inactive storage precursor (1-alkyl-2-acyl-sn-glycero-3-phosphocholine) from 1-alkyl-2-acetyl-sn-glycerol by rabbit platelets.

We have established previously that 1-alkyl-2-acetyl-sn-glycerol (alkylacetyl-G) can be converted into at least six metabolites by rabbit platelets, including alkylacetyl-sn-(glycero-3-phosphocholine) (-GPC), i.e. platelet-activating factor (PAF) and 1-alkyl-2-acyl-sn- (alkylacyl)-GPC. Since part of the biological functions of alkylacetyl-G can be explained by its metabolic conversion to PAF and also to alkylacyl-GPC as an inactive storage precursor of PAF, the present study focused on the regulation of the synthesis of PAF and alkylacyl-GPC from alkylacetyl-G. Our results document the presence of a specific dithiothreitol (DTT)-insensitive cholinephosphotransferase in saponin-permeabilized rabbit platelets and show that DTT potentiates the production of PAF from alkylacetyl-G but inhibits the formation of phosphatidylcholine from diolein. We also demonstrated that the availability of CDP-choline controls the generation of PAF from alkylacetyl-G. Furthermore, when CTP: phosphocholine cytidylyltransferase is activated to produce more CDP-choline through the translocation of this enzyme from the cytosol to membranes by incubating the rabbit platelets with 0.2 mM sodium oleate, the production of PAF from alkylacetyl-G is increased 5-fold. More importantly, our experiments reveal the presence of two metabolic pathways that are responsible for the synthesis of alkylacyl-GPC from alkylacetyl-G, with each producing a unique molecular species composition of the stored PAF precursor, alkylacyl-GPC. The latter is enriched in polyunsaturates (70.7-78.5% 20:4) when formed through the remodeling pathway of PAF cycle via alkylacetyl-G (DTT-insensitive cholinephosphotransferase)----alkylacetyl-GPC----alkyllyso-GPC---- alkylacyl-GPC . Alkylacyl-GPC containing saturated species (71.8% 16:0) is generated by the retroconversion/de novo pathway according to the reaction scheme of alkylacetyl-G----alkyl-G----alkyllyso-glycero-3-phosphate (-GP)----alkylacyl-GP----alkylacyl-G (DTT-sensitive cholinephosphotransferase)----alkylacyl-GPC. Inactivation of PAF through the remodeling/PAF cycle can generate alkylacyl-GPC at both low (1.75 x 10(-7) M) and high (10(-6) M) concentrations of PAF whereas the conversion of alkylacetyl-G to alkylacyl-GPC via PAF through the remodeling pathway only occurs at a low concentration (1.75 x 10(-7) M). At a high concentration (10(-6) M), alkylacetyl-G is converted to alkylacyl-GPC via the retroconversion/de novo route. These data suggest that the formation of PAF by the DTT-insensitive cholinephosphotransferase activity limits the amounts of alkylacyl-GPC produced from alkylacetyl-G through this remodeling pathway (PAF cycle).(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1986

Plasmalogen biosynthesis in Madin-Darby canine kidney cells: selectivity in the acylation of 1-alkyl-2-lyso-sn-glycero-3-phosphoethanolamine and the subsequent desaturation step.

Acyl group specificity in the acylation of 1-alkyl-2-lyso-sn-glycero-3-phosphoethanolamine (1-alkyl-2-lyso-GroPEtn) to form 1-alkyl-2-acyl-sn-glycero-3-phosphoethanolamine (1-alkyl-2-acyl-GroPEtn) and the subsequent desaturation of 1-alkyl-2-acyl-GroPEtn to form plasmalogens (1-alk-1'-enyl-2-acyl-sn-glycero-3-phosphoethanolamine, i.e., 1-alk-1'-enyl-2-acyl-GroPEtn) was investigated in intact Madin-Darby canine kidney (MDCK) cells and cell-free membrane preparations. We found 1-[3H]alkyl-2-lyso-GroPEtn was selectively acylated with polyunsaturated fatty acids in the order 20:4 greater than 20:5 greater than 20:3 (n-9) greater than 22:6 by cell-free membrane preparations of MDCK cells. The same pattern of acyl specificity was seen in intact MDCK cells, although the intact cells produced significantly larger amounts of 1-[3H]alkyl-2-acyl-GroPEtn containing oleic acid. There was an increased desaturation of the 1-[3H]alkyl-2-acyl-GroPEtn species containing docosahexaenoic acid to plasmalogens (1-[3H]alk-1'-enyl-2-acyl-GroPEtn) by both intact MDCK cells and the cell-free membrane preparations. The relatively rapid disappearance of the 1-[3H]alk-1'-enyl-2-docosahexaenoyl-GroPEtn species during a 20-h incubation of prelabeled intact MDCK cells suggests a more rapid turnover of this molecular species. Our results indicate there is a high selectivity in the final acylation and desaturation steps of the biosynthetic pathway for plasmalogens.

Acylation↗
PubMed1984

Novel quantitative method for determination of molecular species of phospholipids and diglycerides.

A novel method is described for the quantitative analysis of subclasses (alk-1-enylacyl, alkylacyl, and diacyl types) and molecular species within each subclass of glycerophosphatides. Diradylglycerols from phospholipase C hydrolysis of the phospholipids are converted to benzoate derivatives, the benzoates are separated into their respective subclasses by thin-layer chromatography, and quantitated by measuring absorbance at 230 nm. Molecular species within individual subclasses are separated using a combination of argentation thin-layer chromatography and reversed-phase high-performance liquid chromatography with direct, on-line quantitation at 230 nm. We applied the method to the analysis of ethanolamine phosphatides from beef brain and were able to quantitate the three diradylglycerol subclasses (alk-1-enylacyl, alkylacyl, and diacyl types) as well as ca. 29 molecular species within each of these subclasses. This new quantitative approach for the analysis of specific molecular species of glycerolipids should be applicable to studies involving a variety of biologically important lipids, such as phosphatidylcholine, phosphatidylinositol, platelet activating factor, plasmalogens, and neutral type glycerolipids including diacylglycerols.

Animals↗
PubMed1982

Formation of alkylacyl- and diacylglycerophosphocholines via diradylglycerol cholinephosphotransferase in rat liver.

We found that diacylglycerol and alkylacylglycerol choline phosphotransferase activities exhibited similar pH optima, thermolabilities and inhibitions by Mn2+, and dithiothreitol. The Vmax of diacylglycerol cholinephosphotransferase was higher (approx. 1-2-fold) than the Vmax of alkylacylglycerol cholinephosphotransferase. The Km value for diacylglycerol was somewhat greater than for the alkylacylglycerol, but no differences were found for the Km of CDPcholine with either type of diradylglycerol as the other substrate. Endogenous levels of diacylglycerols in microsomes were 24.3 nmol/mg protein, whereas no detectable amount of alkylacylglycerols was observed. Results from this study indicate that a similar or identical enzyme catalyzes the formation of 1,2-diacyl- and 1-alkyl-2-acyl-sn-glycero-3-phosphocholine and that the availability of diradylglycerols and the turnover rate of ether-linked lipids would appear to be important factors in controlling the level (under 1%) of 1-alkyl-2-acyl-sn-glycero-3-phosphocholine in rat liver.

Animals↗
PubMed1979

Hypertension screening in schools: results of the Dallas study.

The purpose of this study was to determine the prevalence of persistent blood pressure elevations in an eighth-grade population composed of three ethnic groups, and to determine the feasibility of using school health facilities for hypertension screening. Blood pressure was recorded in 10,641 subjects (90% of the total eighth-grade population) in the Dallas Independent School District. Blacks made up 46% of the population; non-Latin whites, 40.1%; and Latin-Americans, 13.9%. On the first blood pressure screening, 8.9% had systolic or diastolic pressures or both at or above the 95th percentile. Of those whose blood pressures were elevated on the first examination, 98.3% were reexamined. After the third examination, 1.2% continued to have systolic hypertension, and 0.37% diastolic hypertension. No student had diastolic pressure above 90 mm Hg on all three examinations. The prevalence of persistent hypertension was similar for the three ethnic groups. Analysis of variation in blood pressure measurements revealed that the school nurses introduced a relatively small increase in variability. These data indicate that although school screening initially identifies large numbers of students as having inconstant pressure elevations, subsequent follow-up examinations show that less than 2% have persistent hypertension.

Adolescent↗
PubMed1979

Response of hypertensive adolescents to dynamic and isometric exercise stress.

Isometric handgrip and dynamic exercise stress tests were performed on 109 hypertensive and 74 normotensive subjects 14 to 17 years old. The hypertensive subjects had resting systolic or diastolic pressures persistently above the 95th percentile on four consecutive examinations. Blood pressures and ECGs were recorded during isometric handgrip (25% maximum effort for four minutes) and bicycle ergometry until the subject was exhausted. The hypertensive subjects increased systolic pressure by an average 16 mm Hg with isometric exercise and 53 mm Hg with dynamic exercise. Control subjects had similar pressure changes, averaging 18 and 54 mm Hg, respectively. During isometric handgrip stress, diastolic pressures increased 12 mm Hg in hypertensive subjects and 18 mm Hg in control subjects. Only two hypertensive adolescents developed systolic pressures exceeding 200 mm Hg during dynamic exercise stress, and none developed systolic pressures above 200 mm Hg during isometric exercise stress. None of the normotensive or hypertensive subjects developed cardiac arrhythmias and the prevalence of ST segment depression during maximal stress was less than 2% in both groups. Therefore, in adolescents with mild to moderate hypertension the risk of developing significant ECG or hemodynamic abnormalities during mild isometric or heavy dynamic exercise is small. We believe the decision to restrict physical activity of an adolescent with elevated pressures should be based on the development of abnormal ST segment depression, cardiac arrhythmias, or excessive blood pressures at the time of exercise stress testing.

Adolescent↗
PubMed1977

Properties of dihydroxyacetone phosphate acyltransferase in the harderian gland.

We have used a microsomal preparation from the pink portion of the rabbit harderian gland to study the properties of dihydroxyacetone-P acyltransferase. The enzymatic activity in the microsomes is latent and is stimulated approximately 20-fold by the addition of detergent to the assay system. Both deoxycholate and cholate stimulated the enzyme at concentrations considerably below the critical micellar concentration of these detergents. The acyltransferase was not solubilized by treatment with these detergent concentrations. In addition, we have shown that the acyltransferase is sensitive to proteolytic digestion in the presence of deoxycholate concentrations that render the microsomal vesicles permeable to the protease, but is insensitive to the protease treatment in the absence of detergent. Collectively, these data indicate that the active site of dihydroxyacetone-P acyltransferase is exposed to the lumenal surface of the microsomal vesicles. This enzyme is distinct from the glycerol-P acyltransferase in that the dihydroxyacetone-P acyltransferase is sensitive to protease digestion only in the presence of deoxycholate, is not inhibited by sulfhydryl reagents, and is not competitively inhibited by sn-glycerol-3-P.

Acyltransferases↗
PubMed1976

Stimulation of the microsomal alkylglycerol monooxygenase by catalase.

Liver cytosol contains a heat-sensitive nondialyzable soluble factor necessary for maximal activity of the alkylglycerol monooxygenase present in rat liver microsomes. In this report, we demonstrate that the stimulatory component in rat liver supernatant is catalase. Catalase functions to protect the enzyme from inactivation by H2O2 in addition to its documented ability to retard the noenzymatic oxidation of the pterin cofactor. The protective effect of catalase on alkylglycerol monooxygenase and on the aromatic amino acid hydroxylase systems indicates that H2O2 sensitivity is a general feature of pterin-dependent hydroxylases.

Animals↗
PubMed1976

Mass spectral identification of 2-(O-acyl)hydroxy fatty acid esters in the white portion of the rabbit Harderian gland.

A major lipid component of white portion of the rabbit harderian gland has been shown to be a mixture of 2-(O-acyl)hydroxy fatty acid esters. The fatty acid moieties in this lipid class are exclusively saturated and range in chain length from C14:0 to C22:0, with C16:0 being the major component (65%). The fatty alcohols are also saturated and composed primarily of C20:0, C21:0, and C22:0 chains. The hydroxy fatty acids are composed of C14:0, C15:0, and C16:0 and mass spectroscopy combined with chemical techniques placed the hydroxyl group at the 2-carbon. 2-(O-acyl)Hydroxy fatty acid esters are not found in the pink portion of the rabbit harderian gland nor have they been reported to occur in harderian glands or other species.

Animals↗