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T Spennetta

Publications and source records attributed to T Spennetta.

11 recordsLinked to original sources
PubMed1996

Acyl CoA and lipid synthesis from ketone bodies by the extramitochondrial fraction of hepatoma tissue.

The compartmentation of the pathway for the synthesis of lipids from ketone bodies was examined in a cell free, post-mitochondrial supernatant fraction of malignant Morris hepatoma 7777 tissue. A fortified supernatant system effectively incorporated radioactive D(-)-3-hydroxybutyrate and acetoacetate into cholesterol and fatty acids, and both ketone bodies were directly converted to their CoA thioesters. Furthermore, a microsome-free (100,000 x g) cytosolic fraction was also able to acylate 3-hydroxybutyrate to 3-hydroxybutyryl CoA. No previous identification of this enzyme activity has been described. These results which characterize a distinct extramitochondrial pathway for conversion of 3-hydroxybutyrate as well as acetoacetate into lipids also suggest the possibility of a previously undetected enzymatic activity for utilization of this ketone body.

3-Hydroxybutyric Acid↗
PubMed1995

Utilization and preferred metabolic pathway of ketone bodies for lipid synthesis by isolated rat hepatoma cells.

Morris hepatoma 7777 cells freshly isolated from highly malignant tumors grown in the hindlimb of buffalo rats actively convert ketone bodies to cholesterol and fatty acids. On the basis of results obtained with (-)-hydroxycitrate, an inhibitor of the ATP citrate lyase enzyme, the metabolic pathway for acetoacetate conversion to lipids is exclusively cytoplasmic, whereas that for 3-hydroxybutyrate involves both extra- and intramitochondrial compartments. Subcellular distribution studies indicated accumulation and compartmentation of 3-hydroxybutyryl CoA primarily in the cytoplasm of hepatoma cells incubated with either ketone body. In contrast, the compartmentation of acetoacetyl CoA is dependent on whether the substrate is acetoacetate or 3-hydroxybutyrate. With acetoacetate, the acetoacetyl CoA is entirely cytoplasmic, whereas with 3-hydroxybutyrate, it is equally divided between the intra- and extramitochondrial compartments. The results are discussed in terms of the known and proposed metabolic pathways for lipid synthesis from ketone bodies, particularly that from 3-hydroxybutyrate.

3-Hydroxybutyric Acid↗
PubMed1992

Effects of high fat-feeding to rats on the interrelationship of body weight, plasma insulin, and fatty acyl-coenzyme A esters in liver and skeletal muscle.

Rats fed a high-saturated fat diet consumed more energy, gained more weight, and displayed hyperinsulinemia (P less than .05) without an elevation in the fasting plasma glucose level, compared with animals on two different high-carbohydrate diets. The total fatty acyl-coenzyme A (CoA) concentration was 18% (P less than .0001) and 46% (P less than .0001) higher in liver and skeletal muscle, respectively, from rats fed the high-fat diet compared with each of the other diet groups. Major long-chain fatty acyl-CoA molecular species of both tissues in high fat-fed rats reflected the fatty acid profile of the diet. Approximately 29%, 21%, and 16% of total liver and skeletal muscle fatty acyl-CoAs were comprised of oleoyl-CoA, palmitoyl-CoA, and stearoyl-CoA, respectively. The amounts of these three fatty acyl-CoA esters were significantly higher in liver and skeletal muscle after high-fat feeding than with the other diet treatments (P less than .0001). In contrast, the concentration of linoleoyl-CoA was lower in both tissues after high-fat feeding (P less than .0001). In rats fed the high-fat diet, plasma insulin levels were significantly correlated with gain in body weight or body weight (r = .80, P less than .001 for insulin and gain in body weight; r = .73, P less than .001 for insulin and body weight). Total fatty acyl-CoA ester content in liver and skeletal muscle was also strongly correlated with the plasma insulin concentration in high fat-fed rats (r = .80, P less than .001 for liver; r = .78, P less than .001 for skeletal muscle).(ABSTRACT TRUNCATED AT 250 WORDS)

Acyl Coenzyme A↗
PubMed1990

Separation and quantitation of long-chain free fatty acids in human serum by high-performance liquid chromatography.

A rapid, simple and highly sensitive reversed-phase high-performance liquid chromatographic method is described for the separation and quantitation of fatty acids in human serum using a very reactive fluorescent labeling reagent, 9-anthryldiazomethane. Quantitative esterification proceeds at room temperature without heat or catalysis. Baseline separation of nineteen select fatty acids from a standard mixture was achieved on two C18-bonded silica columns connected in tandem using stepwise gradient elution of an acetonitrile-methanol-water mobile phase. The eluent was monitored by a fluorescence detector (maximum excitation wavelength, 365 nm; maximum emission wavelength, 412 nm). The procedure was applied to the analysis of both saturated and unsaturated long-chain free fatty acids (C8 to C22) extracted from human serum. Sera from fasting and non-fasting subjects were analyzed to show the applicability of this assay to biological samples. Detection limit and recovery of free fatty acids in serum were less than 10 pmol/microliter and greater than 92%, respectively.

Adult↗
PubMed1989

Effect of dietary fat on individual long-chain fatty acyl-CoA esters in rat liver and skeletal muscle.

The effect of dietary fat on the long-chain acyl-CoA ester profile of liver and skeletal muscle was investigated by feeding weanling rats 12%-fat diets composed of high-linoleic safflower oil (73% 18:2n-6), high-oleic safflower oil (70% 18:1n-9) or olive oil (70% 18:1n-9) for six and ten weeks. Approximately 50% of both hepatic and skeletal muscle acyl-CoA esters comprised linoleoyl-CoA or oleoyl-CoA with high-linoleic or oleic feeding, respectively. Total hepatic acyl-CoA ester concentration was 40% higher (p less than 0.05) in rats fed 12% fat compared with controls fed a 4%-fat diet. These data demonstrate that the long-chain acyl-CoA ester profile of liver and skeletal muscle reflects the dietary fatty acid profile.

Acyl Coenzyme A↗
PubMed1988

Adaptive changes in individual acyl-CoA esters from hamster BAT during cold acclimation.

Long-chain fatty acyl-CoA esters (LCFACoAE) were extracted from freeze-clamped powdered brown adipose tissue (BAT) obtained from thermoneutral control and cold-acclimated hamsters and the CoA esters individually separated by high-performance liquid chromatography. LCFACoAE of carbon chain length C12 to C20 were identified by increasing column retention time in the following order: C12:0, C14:1, C14:0, C16:1, C18:2, C16:0, C18:1, C18:0, and C20:4. The mean total LCFACoAE concentrations were 235 +/- 40 nmol/g protein for the control hamsters and 648 +/- 105 nmol/g protein for the 22-day cold-acclimated hamsters. A rapid fourfold increase in the levels of C16:0, C18:0, and C18:1 occurred within hours after initiation of the cold temperature, whereas the concentrations of the other six LCFACoAE either increased only slightly or remained unchanged. Almost 50% of the total LCFACoAE in the BAT of cold-acclimated hamsters was made up of C16:0, C18:0, and C18:1. These results, which demonstrate some dynamic changes in adipose tissue LCFACoAE, are consistent with their proposed role in the initiation and maintenance of BAT thermogenesis.

Acclimatization↗
PubMed1985

Extraction of tissue long-chain acyl-CoA esters and measurement by reverse-phase high-performance liquid chromatography.

Long-chain acyl-CoA esters were extracted from freeze-clamped livers of fed and fasted rats according to the method of Mancha et al. [M. Mancha, G. B. Stokes, and P. K. Stumpf (1975) Anal. Biochem. 68, 600-608] and analyzed on a radially compressed C18, 5 microns, reverse-phase column using a gradient system consisting of acetonitrile and 25 mM KH2PO4, pH 5.3, at 254 nm. Total analysis time was 25 min. Eight peaks in the extract with carbon chain lengths of 12 to 18, which subsequently disappeared on alkaline hydrolysis, were identified. The major acyl-CoA peaks in the extract in order of increasing retention times were 14:0, 16:1, 18:2, 16:0, 18:1, and 18:0. Total liver long-chain acyl-CoA esters were 108 +/- 11 and 248 +/- 19 nmol/g protein for fed and fasted rats, respectively. On fasting (48 h) the levels of 18:2, 16:0, and 18:1 increased two-to threefold and that of 18:0 sixfold. The advantages of this method are that it not only provides a more direct determination of total tissue long-chain acyl-CoA esters, in that no decomposition of the CoA ester is involved, but it also detects the constituent molecular species.

Acyl Coenzyme A↗
PubMed1983

Total cholesterol and HDL-cholesterol changes during acute, moderate-intensity exercise in men and women.

Chronic endurance exercise training has been associated with decreased levels of total cholesterol and increased HDL-cholesterol. To our knowledge rapid changes in cholesterol and HDL-cholesterol during acute exercise have not been described under controlled conditions. We studied 28 subjects (14 males and 14 females) during bicycle exercise for 40 min at a work intensity of 55% of their maximal oxygen consumption. Total and HDL-cholesterol levels were measured (and LDL-cholesterol calculated) at rest, 10, 20, 30, and 40 min of exercise, and 15 min postexercise. There was a significant (p less than 0.001) increase in HDL-cholesterol levels at 10 min of exercise (58.8 +/- 13.9 mg/dl, mean +/- SD) above rest (53.1 +/- 13.4 mg/dl) for all subjects. This increase persisted (p less than 0.001) at all time points throughout the exercise session, but declined by 15 min postexercise. There was a small, insignificant decline in LDL-cholesterol. It is concluded that apparent favorable changes in lipoprotein patterns occur acutely, and are sustained during short-term, moderate intensity exercise. Analyses of these changes appears necessary if the biochemical mechanisms which underlie these metabolic alterations are to be elucidated.

Adolescent↗
PubMed1976

The carbon pathway for lipogenesis in isolated adipocytes from rat, guinea pig, and human adipose tissue.

The synthesis of fatty acids from a variety of labeled substrates by isolated adipocytes of the rat, guinea pig, and human was investigated. The incorporation of radioactive glucose and pyruvate into triglyceride fatty acids was considerably lower in human than either rat or guinea pig adipose tissue. By contrast, the incorporation of palmitate into adipose tissue triglycerides was approximately the same in all three species. End carbon analysis of fatty acids isolated from adipocytes incubated with pyruvate-U-14C indicated that although the synthesis of fatty acids in human adipose tissue was extremely low compared to that of the rat and guinea pig, it represented de novo biosynthesis rather than chain elongation of existing fatty acids. It is suggested that in the human, fatty acids are synthetised de novo primarily in the liver. In adipose tissue, lipogenesis consists essentially of the esterification of fatty acids, obtained from plasma, into triglycerides.

Adipose Tissue↗