Intramitochondrial reductive carboxylation of 2-oxoglutarate in adipose tissue and its contribution to fatty acid synthesis

• 1.1. The reductive carboxylation of 2-oxoglutarate was found to proceed in mitochondria of rat epididymal fat pads and rabbit perirenal adipose tissue at a rate similar to that in liver mitochondria.
• 2.2. In rat fat pads the incorporation of ¹⁴C from [5-¹⁴C]2-oxoglutarate into fatty acids via the carboxylation was suppressed by butylmalonate by 30%.
• 3.3. 2-Oxoglutarate and glutamate stimulated the incorporation into fatty acids of ¹⁴C from [2-¹⁴C]acetate in rat fat pads with the simultaneous reduction of tissue NADP. These effects persisted after inhibition of succinate dehydrogenase by malonate.
• 4.4. It is concluded that in adipose tissue 2-oxoglutarate carboxylation proceeds in both the cytoplasm and mitochondria. Therefore, it can supply carbon atoms as well as NADPH for fatty acid synthesis.

     

Influence of dietary sources of fat on lipid synthesis in mink (Mustela vison) mammary tissue

• 1.1. The fatty acid composition of the triglyceride fraction of mink milk sampled during mid-lactation (day 28 post partum) from two nursing mink was compared to that of plasma samples and to the fatty acid composition of the feed rations used.
• 2.2. Chemical analysis of the triglyceride composition of mink milk demonstrated only minute concentrations of fatty acids with a chain length below C₁₄.
• 3.3. The saturated C_16:0- and C_18:0-unit fatty acids in mink milk made up for 24–40% of the total amount of fatty acids extracted, the remainder being represented by mono and polyunsaturated long-chain (C₁₆-C₂₄) fatty acids.
• 4.4. Preliminary in vitro experiments proved the incorporation of¹⁴C-labelled glucose, acetate or palmitate into triacylglycerols in cultures of mink mammary tissue to be linear for at least 2 hr.
• 5.5. The in vitro capacity for de novo fatty acid synthesis in mink mammary tissue using 14C-labelled glucose or acetate was low, i.e. ranging from 0.096–0.109 nmol/g (fresh tissue)/min, and amounted to only about 5% of that obtained in the case of [¹⁴C]palmitic acid incubation.
• 6.6. Following ¹⁴C-labeIled acetic or palmitic acid incubation of mink mammary tissue neither desaturation nor chain elongation was observed.
• 7.7. In response to long-term feeding on rations with two different sources of animal fat (F = fish oil or L = lard) the influence of compositional changes in dietary neutral lipids on the fatty acid composition of the lipids of mink milk is discussed.

     

Modulation of mitochondrial ATPase sensitivity to inhibitors and stimulators by diet-induced changes in membrane lipid

• 1.1. Weanling rats were fed diets differing in fatty acid composition to determine if changes induced in cardiac mitochondrial membrane structural components alter the sensitivity of mitochondrial ATPase to inhibition by oligomycin and stimulation by 2,4-dinitrophenol.
• 2.2. Mitochondrial ATPase assayed in situ within the mitochondrial membrane isolated from animals fed diets higher in fatty acids of longer chain length, exhibited greater oligomycin sensitivity and lower 2,4-dinitrophenol-induced stimulation.
• 3.3. Concomitant diet-induced changes occur in the fatty acid, composition of phosphatidylcholine, phosphatidylethanolamine and cardiolipin, increasing overall length of fatty-acyl tails in the membrane phospholipids.
• 4.4. Diet fat mediated alterations in oligomycin sensitivity of mitochondrial ATPase and membrane fatty acid chain length suggest that _vivo changes in thickness of the lipid bilayer may alter mitochindrial ATPase functions.
• 5.5. The present study extends the concept that dietary fat affects mitochondrial membrane structure and function by demonstrating that the membrane-dependent sensitivity of mitochondrial ATPase to inhibitors and stimulators may be modulated by dietary fat.

     

Effects of milk fat,unhydrogenated and partially hydrogenated vegetable oils on serum lipoproteins in growing pigs

• 1.1. Crossbred Yorkshire (Yorkshire × Landrace) pigs were fed butter oil, cream, low erucic acid rapeseed oil, sunflower oil and partially hydrogenated sunflower oil in amounts representing 30% of energy for periods of up to 13 weeks.
• 2.2. After 13 wk of feeding serum total cholesterol levels of pigs fed milk fat were significantly higher than of pigs fed vegetable oils.
• 3.3. The difference in cholesterol was mainly due to an increase in the density range of 1.063–1.125 g/ml containing pig LDL₂ and some HDL.
• 4.4. A shift towards smaller LDL particle size was apparent in pigs fed milk fat.
• 5.5. The effects of dietary trans fatty acids did not differ from cis polyunsaturated or monounsaturated fatty acids.

     

Differential effects of fatty acid chain length on the viability of two species of cactophilic Drosophila

• 1.1. Two columnar cacti in the Sonoran Desert, agria and organpipe, contain medium chain (C₈−C₁₂) fatty acids.
• 2.2. Necrotic tissues of these cacti serve as feeding and breeding substrates for Drosophila mojavensis but not D. nigrospiracula.
• 3.3. Results show that capric and lauric acids are the predominant fatty acids of both cacti.
• 4.4. Fatty acid chain length exhibits a differential effect on larval viability with caprylic acid (Q) having the greatest and myristic acid (C₁₄) having the least effect.
• 5.5. Drosophila mojavensis is more tolerant of free fatty acids than D. nigrospiracula, and this partly explains the ability of D. mojavensis to utilize agria and organpipe cacti.

     

Purification and some properties of isocitrate dehydrogenase of a blowfly Aldrichina grahami

• 1.1. NADH-dependent isocitrate dehydrogenase has been purified 110-fold from the crude extract of the flight muscle mitochondria of Aldrichina grahami.
• 2.2. The purification procedure involved Triton X-100 treatment of isolated mitochondria, column chromatography on DEAE-cellulose, Affi-gel blue, and P-cellulose.
• 3.3. The purified enzyme was homogeneous by criteria of the polyacrylamide gel electrophoresis.
• 4.4. The enzyme of the blowfly contains more acidic amino acids and less hydrophobic amino acids than that of pig heart.
• 5.5. The molecular weight was determined to be 330,000 daltons. The subunit construction differs from ghat of mammalian isocitrate dehydrogenase.

     

Sterols from black sea invertebrates—I. Sterols from Scyphozoa and Anthozoa (Coelenterata)

• 1.1. Rhizostoma pulmo, Aurelia aurita and Actinia equina, the most widespread representatives of Coelenterata in Black Sea have been analysed and the occurrence of 20 sterols has been found.
• 2.2. Dinosterol and demethyldinosterol as well as a number of short side chain sterols have been found in Scyphozoa for the first time.
• 3.3. The occurrence of coprostanol in marine invertebrates has been shown for the first time.
• 4.4. Five groups of sterol esters were found, containing fatty acids with different polarity.

     

Fermentation and the production of volatile fatty acids in the alimentary tract of the rock hyrax,Procavia capensis

• 1.1. Aspects of ruminant-like metabolism were examined in the hyrax Procavia capensis.
• 2.2. High concentrations of volatile fatty acids occurred in the cardiac stomach with a predominance of acetic and lactic acids.
• 3.3. Acetic (69%), propionic (22%) and butyric (8%) acids occurred in highest concentrations in the proximal caecum, with appreciable amounts in the proximal colon, distal caecum and appendices.
• 4.4. The depot fat contained high proportions of unsaturated C₁₈ (linoleic and linolenic) acids.
• 5.5. The glucose level in the plasma was within the range established for non-ruminant herbivores.
• 6.6. The possibility of silage-like fermentation occurring in the cardiac stomach is discussed.

     

Changes in plasma glucose and lipid concentrations during treadmill exercise in domestic fowl

• 1.1. Plasma glucose, non-esterified fatty acid, triglyceride, cholesterol and lactate concentrations were measured during 90 min treadmill exercise at a work intensity of 55–60% maximum.
• 2.2. After 90 min exercise plasma glucose fell by 35% whilst the non-esterified fatty acid concentration rose to as much as 3–4 times resting.
• 3.3. Exercise had no significant effect on plasma cholesterol, triglyceride or lactate concentrations.
• 4.4. The findings indicate a progressive increase in fat utilization during prolonged exercise. Possible hormonal mechanisms underlying exercise-induced changes in lipid and carbohydrate metabolism are discussed.

     

Role of glutamate dehydrogenase reaction in the control of citrate pool in yeast

• 1.1. Role of NADP-glutamate dehydrogenase in the depletion of citrate was analyzed using permeabilized yeast cells.
• 2.2. Citrate was converted to 2-oxoglutarate, which was then metabolized to glutamate by NADP-glutamate dehydrogenase in the presence of ammonium ion.
• 3.3. Formation of 2-oxoglutarate plus glutamate was in good agreement with the concentration of citrate decreased. Glutamate formation can be a good indicator of the depletion of citrate, because 70% of the citrate decreased was converted to glutamate.
• 4.4. Glycolytic activity was closely correlated with the decrease in citrate under the in situ conditions.
• 5.5. NADP-glutamate dehydrogenase increased in anaerobically grown yeast cells.
• 6.6. An effective depletion of citrate by increased synthesis of NADP-glutamate dehydrogenase can explain the lowered mechanism of citrate causing glycolytic stimulation under the anaerobic growth conditions of yeast.

     

Changes in blood metabolite concentrations in response to repeated capture,anaesthesia and blood sampling in the golden perch,Macquaria ambigua

• 1.1. The major metabolic changes associated with repeated capture, aquarium transfer, anaesthesia and blood sampling were investigated in an Australian freshwater fish, the golden perch (Macquaria ambigua),
• 2.2. A compounded stress response was seen after repetition of the procedure, in which the plasma glucose rose within 3 hr and amino acid concentrations rose and the serum free fatty acids concentration fell after 24 hr.
• 3.3. Alanine was identified as an important circulating energy store in the stress response of golden perch.
• 4.4. No change was noted in the serum protein, plasma lactate or β-hydroxybutyrate concentrations, indicating that tissue damage and hypoxia were absent, and that degradation of free fatty acids did not produce metabolites excess to the requirements of gluconeogenesis and the tricarboxylic acid cycle.

     

A survey of the non-esterified fatty acids and binding proteins in the plasmas of selected animals

• 1.1. The levels of non-esterified fatty acids (NEFA) in the plasma of a variety of animals have been estimated.
• 2.2. Only one of seven elasmobranchs contained detectable levels of NEFA.
• 3.3. The two crustaceans examined contained very low levels.
• 4.4. All the other animals contained circulating levels of a variety of NEFA ranging from 14 to 24 carbon atoms.
• 5.5. The elasmobranchs are unique in that they also do not possess proteins in the serum which bind fatty acids.

     

Changes in the phospholipid composition and in fatty acids of phosphatidylcholine and phosphatidylethanolamine of various tissues of the developing tadpole of Agalychnis dacnicolor

• 1.1. Lipid changes occur in the developing tadpole of A. dacnicolor. The phosphatidylcholine content of liver and tail decrease during metamorphosis.
• 2.2. In liver, the fatty acids of phosphatidylcholine and phosphatidylethanolamine become more unsaturated.
• 3.3. In skin, phosphatidylcholine becomes more unsaturated and phosphatidylethanolamine becomes more saturated.
• 4.4. In tail, phosphatidylcholine becomes more saturated and phosphatidylethanolamine shows no change.
• 5.5. Triglycerides become more unsaturated in skin but become more saturated in tail.

     

Lipid digestion in cod (gadus morhua)

• 1.1. Cod, 2.6–3.4 kg. were fed a mixed diet of sprat, capelin oil and wheat flour.
• 2.2. Lipids from the feed, stomach and four intestinal segments were separated into tri-, di- and monoglycerides and free fatty acids and analysed by GLC.
• 3.3. All lipolytic products were concentrated in 14:0, 16:0 and 18:0, up to 60% and extremely low in the ω-3 fatty acids.
• 4.4. Residual triglycerides contained 80% of saturated and monoenoic fatty acids.
• 5.5. Linoleic acid increased from 2% in feed TG to 10% in TG of the rectum.

     

A new glutamate dehydrogenase from Halobacterium halobium with different coenzyme specificity

• 1.1. Halobacterium halobium has two chromatographically distinct forms of glutamate dehydrogenase which differ in their thermolability and other properties. One glutamate dehydrogenase utilizes NAD, the other NADP as a coenzyme.
• 2.2. The NADP-specific glutamate dehydrogenase (EC 1.4.1.4) was purified 65-fold from crude extracts of H. halobium.
• 3.3. The Michaelis constants for 2-oxoglutarate (13.3 mM), ammonium (3.1 mM) and NADPH (0.077 mM) indicate that the enzyme catalyzes in vivo the formation of glutamate from ammonium and 2-oxoglutarate.
• 4.4. The amination of 2-oxoglutarate by NADP-specific glutamate dehydrogenase is optimal at the pH value of 8.0–8.5. The optimal NaCl or KCl concentration for the reaction is 1.6 M.
• 5.5. None of the several metabolites tested for a possible role in the regulation of glutamate dehydrogenase activity appeared to exert an appreciable influence on the enzyme.
• 6.6. NAD- and NADP-dependent glutamate dehydrogenases from H. halobium showed apparent molecular weights of 148,000 and 215,000 respectively.

     

Relations between fatty acid synthesis,pyruvate concentration and cell concentration of suspensions of isolated rat hepatocytes

• 1.1. The cell concentration of suspensions of isolated rat hepatocytes affects both the rate of pyruvate accumulation in the incubation medium and the rate of fatty acid synthesis.
• 2.2. At low cell concentrations pyruvate accumulation is directly related to the cell concentration but levels off at higher concentrations even when maximum pyruvate concentrations in the medium are not yet reached.
• 3.3. The rate of fatty acid synthesis in the 30–60-min incubation interval is proportional to the cell concentration. In contrast, the rate of fatty acid synthesis during the 0–30-min incubation period decreases with increasing cell concentrations and subsequently becomes independent of the cell concentration.

     

Effect of different levels of dietary fats upon the positional distribution of fatty acids in rat body triacylglycerols

• 1.1. The main purpose of the study was to describe and to compare the effect of different dietary fatty acids introduced at different levels into the diet on the positional distribution of fatty acids in rat body triacylglycerols. Only distribution between sn-2 (internal) and sn-1 + sn-3 (external) positions was considered in this study.
• 2.2. The positional distribution of fatty acids was first determined for controls fed on a low fat diet (1% D.M.). The same study was then carried out with 11 different dietary treatments and results were systematically compared with controls. The effects of four main molecules were tested: linoleic acid, oleic acid, lauric acid and capric acid.
• 3.3. Different alterations of the positional distribution of fatty acids in body triacylglycerols were obtained. They are graphically described and discussed.

     

The intracellular compartmentation of metabolites in isolated kidney cortex tubules

• 1.1. A method is described to measure cytosolic and mitochondrial metabolites in isolated kidney tubule suspensions.
• 2.2. With the use of digitonin, 95% of cytosolic metabolites are released into the supernatant fraction whereas mitochondrial matrix enzymes are retained in the pellet fraction.
• 3.3. In a study of adaptation to acidosis, different α-oxoglutarate gradients between the tubular cell compartments are obtained, when medium pH is changed from 7.0 to 7.8.

     

The influence of seasonal changes on energy metabolism in Mytilus edulise (L.).—III. Anaerobic energy metabolism

• 1.1. Seasonal changes in the accumulation of end products after 48 hr of exposure to air and in the composition of the free amino acid pool were studied in Mytilus edulis.
• 2.2. The accumulation levels of succinate and acetate showed only weak seasonal changes.
• 3.3. Conversion of succinate to propionate was high in summer and virtually zero in winter
• 4.4. Alanine and most other free amino acids were present in relatively high concentrations in summer and early autumn and reached minimal values in winter and early spring.
• 5.5. Exceptions were glutamate, aspartate and taurine, which showed hardly an season related changes and glycine, which changed inversely to the majority of the free amino acids.
• 6.6. The anaerobic formation of alanine was inversely proportional to the endogenous concentration.
• 7.7. The only other free amino acids affected by anaerobiosis were glutamate and aspartate, which respectively increased and decreased under these conditions.

     

Phospholipid and fatty acid composition of the freshwater sponge Euspongilla lacustris from the volga river estuary

• 1.1. Phospholipids of the freshwater sponge Euspongilla lacustris from the Volga river estuary were examined.
• 2.2. The freshwater sponges belonging to the family Spongillidae were shown to contain demospongic fatty acids.
• 3.3. Composition of fatty acids in phospho-, glyco- and neutral lipid fractions was studied.