[The decomposition of urea in Ferguson synthetic medium by Past. pseudotuberculosis, new reaction for differentiating the etiologic agents of plague and pseudotuberculosis].
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Fatty Acids↗
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Results for “FATTY ACIDS, RADIOACTIVE”
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Relative significance of exogenous and de novo synthesized fatty acids in the formation of rumen microbial lipids in vitro.
Mixed rumen microorganisms (MRM) or suspensions of rumen Holotrich protozoa obtained from a sheep were incubated anaerobically with [1-(14)C]linoleic acid, [U-(14)C]glucose, or [1-(14)C]acetate. With MRM, the total amount of fatty acids present did not change after incubation. An increase in fatty acids esterified into sterolesters (SE) and polar lipids at the expense of free fatty acids was observed. This effect was intensified by the addition of fermentable carbohydrate to the incubations. Radioactivity from [1-(14)C]linoleic acid was incorporated into SE and polar lipids with both MRM and Holotrich protozoa. With MRM the order of incorporation of radioactivity was as follows: SE > phosphatidylethanolamine > phosphatidylcholine. With Holotrich protozoa, the order of incorporation was phosphatidylcholine > phosphatidylethanolamine > SE. With MRM the radioactivity remaining in the free fatty acids and that incorporated into SE was mainly associated with saturated fatty acids, but a considerable part of the radioactivity in the polar lipids was associated with dienoic fatty acids. This effect of hydrogenation prior to incorporation was also noted with Holotrich protozoa but to a much lesser extent. Small amounts of radioactivity from [U-(14)C]glucose and [1-(14)C]acetate were incorporated into rumen microbial lipids. With protozoa incubated with [U-(14)C]glucose, the major part of incorporated radioactivity was present in the glycerol moiety of the lipids. From the amounts of lipid classes present, their radioactivity, and fatty acid composition, estimates were made of the amounts of higher fatty acids directly incorporated into microbial lipids and the amounts synthesized de novo from glucose or acetate. It is concluded that the amounts directly incorporated may be greater than the amounts synthesized de novo.
Acetates↗
Metabolism of liver triacylglycerols in rats tube-fed a threonine-devoid diet.
Rats tube-fed a diet devoid of threonine accumulated triacylglycerols in their livers, starting on the third day of the diet. The fatty acid composition of the accumulated lipid and the contribution of novo synthesized fatty acids to the lipid accumulation, as determined with tritiated water as a radioactive precursor for fatty acid synthesis, suggested that an increased hepatic de novo synthesis of fatty acids is not a major factor for the development of this liver lipid accumulation. The metabolism of intravenous injected 3H-oleic acid, the Triton-induced hyperlipemia and the activity of lipoprotein lipase in adipose tissue was also studied. None of these studies revealed any significant difference between the threonine-deficient and control rats. It is concluded that the hepatic triacylglycerol accumulation in the threonine-deficient rats does not result from any gross abnormality in the rate of liver triacylglycerol formation or secretion to the plasma. It is suggested that a possible causative mechanism is a derangement in the metabolism of the storage pool of liver triacylglycerols.
Adipose Tissue↗
Comparative metabolism of erucic and oleic acid in hepatocytes from rats fed partially hydrogenated marine oil or palm oil.
Rats were fed for 8 weeks high fat diets containing 20% of either palm oil (PO) or linoleic-supplemented partially hydrogenated marine oil (PHMO). Hepatocytes were isolated from rats fasted overnight. Incubations were performed with albumin bound [10-14C] oleic acid and [14-14C] erucic acid, respectively, for 15 or 60 min. The cell viability was better than 90% after 60 min of incubation. The incorporation of radioactive metabolites into lipids of the cells and of the secreted products was determined. The uptake and channelling of 18:1 and 22:1 into various lipid classes in the hepatocytes were different, but were little influenced by the diets. Erucic acid disappeared more rapidly from the incubation medium than 18:1 at both incubation times and for both dietary groups, and was mainly incorporated into the hepatocyte triglycerides and found in the free fatty acid pool, whereas only small amounts of labelling were detected in the phospholipids. Oleic acid, however, was readily incorporated into both phospholipids and triglycerides, and little was found in the free fatty acid pool. For both the triglycerides and the free fatty acids more than 90% of the radioactivity was represented by the incubation fatty acid. Chain-shortened products were found in the phospholipids, especially after incubation with 22:1. The chain-shortening capacity was somewhat higher in the PHMO group, relative to the PO group. At both incubation times, the major part (80-90%) of the total radioactivity was found in the lipid-soluble fraction of the incubation medium, and for both groups 60-80% still as the original incubation fatty acid. 20-30% of the radioactivity was found in the triglycerides secreted from the cells. Phospholipids were secreted in minor amounts (1-5%). Water soluble products constituted 3-7% at 15 min of incubation, increasing to 10-12% at 60 min, highest when incubating with 22:1 and similar for both dietary groups.
Animals↗
A micro quantitative determination of total fatty acids in biological samples by esterification with radioactive methanol.
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Cholesterol↗
Myelin proteolipid protein contains thioester-linked fatty acids.
Myelin proteolipid protein (PLP) is known to contain long-chain, covalently bound fatty acids. Previous studies, including our own, have suggested the occurrence of an oxyester type of linkage between fatty acids and PLP. However, we found that protein-SH groups are required in the acylation reaction, suggesting the possible presence of thioesters. In the present study, we have examined the nature of the acyl-PLP linkages by determining whether free thiol groups are generated on removal of fatty acids. Incubation of reduced and carboxyamidomethylated proteolipid apoprotein (RCM-APL) with 0.2 M hydroxylamine and [14C]iodoacetamide at pH 7.5 and 37 degrees C resulted in the release of fatty acids and the concomitant labeling of newly formed thiol groups. Incubation with Tris or methylamine at pH 7.5 failed to remove fatty acids and generate free -SH groups. The possibility that on treatment buried thiol groups became exposed was essentially excluded because (1) similar results were obtained in 2-chloroethanol, a solvent in which acylated and deacylated PLP have the same conformation, and (2) small PLP peptides were labeled only in the presence of hydroxylamine. On incubation with [14C]methylamine at pH 9.0, RCM-APL was not labeled, thus excluding the occurrence of intramolecular thiol esters. On the other hand, fatty acids were released as radioactive N-methyl fatty acylamide, indicating the presence of intermolecular thioesters between fatty acids and protein. These results demonstrate that a large proportion of fatty acids covalently bound to PLP are liked to -SH groups.
Acylation↗
Anti-hypertriglyceridaemic activity of some phenylethylamine anorectic compounds.
In rats allowed access to food, and in food-deprived rats, fenfluramine (20 and 100 mg kg-1) and amphetamine (10 and 20 mg kg-1) provoked a hypotriglyceridaemic effect. No changes in plasma cholesterol concentration were observed. The time course of the absorption of a lipid load differed according to the nutritional status of the animals; being bellshaped under fed, and curvilinear under fasted, conditions. However, absorption under both nutritional conditions was inhibited by amphetmine and fenfluramine. When rats which had received the test compounds were administered glycerol trioleate containing a tracer dose of glycerol [1-14C]-trioleate or [2-3H]-glycerol trioleate, there was an inhibition in the increase of plasma radioactivity only in the case when the fatty acid contained the radioactive label. The net effect of lipid absorption was a transfer of dietary lipid from the gut to adipose tissue stores. There was never more than 5 per cent of the administered load in the liver. These observations indicate that amphetamine and fenfluramine may have acute effects in reducing circulating triglycerides, separate from the effects on lipid absorption from the gut. In this latter, the palmitoyl-CoA monooleinacyltransferase enzyme probalby plays a key role and appears a major target of the overall anti-obesity of fenfluramine.
Animals↗
Incorporation of polyunsaturated fatty acids into lipids of rainbow trout hepatocytes.
Hepatocytes from 5- or 20 degrees C-acclimated rainbow trout (Salmo gairdneri) were incubated with [1-14C]oleic, -linoleic, or -linolenic acid. Both acclimation groups demonstrated greater incorporation of derivatives from linolenic and linoleic acids into phospholipids when assayed at 5 and 20 degrees C; few derivatives of oleic acid were formed. Cells from cold-acclimated trout, when assayed at 5 degrees C with linolenic acid, incorporated a large proportion of radioactivity into free fatty acids. Analysis of each lipid fraction revealed a relatively specific incorporation of certain fatty acids. For example, "dead end" elongation products of the three substrates were preferentially incorporated into neutral lipids, while delta 6 desaturation products of the three acids were retained in the free fatty acid fraction. Twenty-carbon acid derivatives of linoleic and linolenic acids were directed into the phospholipid fraction. Incorporation of the delta 5 desaturation products was temperature sensitive in cells from cold-acclimated but not warm-acclimated trout. The results suggest that selectivity of incorporation of specific fatty acids into phospholipids may be of importance in restructuring membranes of poikilotherms during thermal adaptation.
Animals↗
Incorporation of 14C acetate into the phospholipids and fatty acids of rabbit lens in organ culture.
Lenses were cultured in Medium 199 containing 14C acetate for 3, 6, 12 and 20 hours. There was a linear increase in incorporation into the total lipids with time. The total lipid extract was subjected to two-dimensional chromatography and incorporation into the individual classes of lipids was measured. For all time periods approximately 70% of all incorporated radioactivity was seen in the sterol and phosphatidyl choline (PC) fractions. Incorporation was observed in phosphatidyl ethanolamine (PE), aldehydes of PC and PE plasmalogens, phosphatidyl inositol (PI), PC plasmalogens, PE plasmalogens, sphingomyelin (SM), phosphatidic acid (PA) and phosphatidyl serine (PS). The fatty acids of the total lipids were methylated and subjected to gas chromatography, and individual fatty acids were collected. The greatest incorporation occurred in palmitic acid (16:0) and myristic acid (14:0) with minor amounts in lauric acid (12:0). These are the major products of de novo synthesis. The remainder of the radioactivity appeared in fatty acids 18 carbons or longer, which are the products of an elongation mechanism.
Acetates↗
Myocardial metabolism of radioiodinated methyl-branched fatty acids.
Methylated fatty acids labeled with radioactive iodine have been proposed as a means of studying regional myocardial uptake of fatty acids in man. To investigate the methylated fatty acid that is best adapted for an assessment of uptake, we have studied the influence of the number and the position of the methyl groups of IFA intracellular metabolism; 16-iodo-2-methyl-hexadecanoic (mono-alpha), 16-iodo-2,2-methyl hexadecanoic (di-alpha), 16-iodo-3-methyl-hexadecanoic (mono-beta), and 16-iodo-3,3-methyl-hexadecanoic (di-beta) acids were injected into the coronary arteries of isolated rat hearts. Intracellular analysis shows that the degradation of mono-alpha was always lower than that of IHA and the storage was always much higher. The differences between mono-beta and IHA were similar to those observed with mono-alpha, but were much more pronounced. With the two dimethylated IFAs there was an inhibition of both oxidation and esterification which led to an accumulation of free FAs in myocardial cells. In conclusion, mono-beta, di-alpha, and di-beta are potentially suitable for studying the cellular uptake of IFA since all of them, and particularly the dimethylated IFAs, have a low oxidation rate.
Animals↗
Preliminary investigations of carbon tetrafluoride as an inert diluent gas to prevent explosions of mixtures of cyclopropane and oxygen.
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Cyclopropanes↗
[Considerations on 25 cases of postabortal hepatonephritis].
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Abortion, Induced↗
Rates and products of long-chain Fatty Acid synthesis from [1-C]acetate in chloroplasts isolated from leaves of 16:3 and 18:3 plants.
Chloroplasts highly active in the synthesis of long-chain fatty acids from [1-(14)C]acetate were prepared from leaves of Solanum nodiflorum, Chenopodium quinoa, Carthamus tinctorius, and Pisum sativum. These preparations were used to test whether the various additions to incubation media found to stimulate the synthesis of particular lipid classes in vitro by Spinacia oleracea chloroplasts were applicable generally. Chloroplasts from 18:3 plants incorporated a greater proportion of radioactivity into unesterified fatty acids under control conditions than did those from 16:3 plants. Supplying exogenous sn-glycerol 3-phosphate or Triton X-100 to chloroplasts increased the synthesis of glycerolipids in all cases and accentuated the capacity of chloroplasts from 18:3 plants to accumulate phosphatidic acid rather than the diacylglycerol accumulated by chloroplasts from 16:3 plants. The UDP-galactose-dependent synthesis of labeled diacylgalactosylglycerol was much less active in incubations of chloroplasts from 18:3 plants also containing sn-glycerol 3-phosphate and Triton X-100 compared with similar incubations from 16:3 plants. Exogenous CoA stimulated total fatty acid synthesis in all chloroplast preparations and the further addition of ATP diverted radioactivity from the unesterified fatty acid to acyl-CoA. The results have been discussed in terms of the two pathway hypothesis for lipid synthesis in leaves.
Journal Article↗
The measurement of production rates of volatile fatty acids in the caecum of the conscious rabbit.
1. The decrease in specific radioactivity of individual volatile fatty acids (VFA) after a single injection of tracer was monitored. The results obtained indicated the occurrence of a first-order process. 2. Regression analysis indicated the high flux of VFA through the caecal pool, equivalent to 30% of the maintenance energy requirement of the animal. 3. Interconversion of VFA was monitored, and results indicated substantial synthesis of butyric acid from acetic acid. 4. Results were obtained from animals on two dietary regimens, and these were compared with results reported for the other species.
Acetates↗
Effect of nerve growth factor on the synthesis of amino acids in PC12 cells.
Radioactive short-chain fatty acids preferentially label glutamine relative to glutamate in brain due to compartmentation of glutamine and glutamate. To determine whether this phenomenon occurs in a single cell culture model, we examined the effect of fatty acid chain length on the synthesis as well as pool size of selected amino acids in rat pheochromocytoma PC12 cells, a cell culture model of the large glutamate compartment in neurons. Intracellular 14C-amino acids were quantitated by HPLC, and the incorporation of [U-14C]-glucose, [1-14C]-butyrate, [1-14C]-octanoate, and [1-14C]-palmitate into five amino acids was measured in native and NGF-treated PC12 cells. NGF pretreatment decreased the intracellular concentration of amino acids as did addition of fatty acids but these effects were not additive. Specific activities of amino acids in native cells labelled by 14C-octanoate were 1,300 DPM/nmol, 490 DPM/nmol, 200 DPM/nmol, and 110 DPM/nmol for glutamate, aspartate, glutamine, and serine, respectively. No radioactivity was detected in alanine. Similar specific activities were noted when 14C-butyrate was the precursor; however, there was at least 5-fold less if 14C-palmitate was the precursor. Pretreatment of cells with NGF decreased the specific activity of amino acids by 25-65%. Specific activities of amino acids synthesized from 14C-glucose decreased in the following order: glutamate, 1,640 DPM/nmol; aspartate, 1,210 DPM/nmol; alanine, 580 DPM/nmol; glutamine, 275 DPM/nmol; and serine, 80 DPM/nmol for native cells. NGF pretreatment decreased the specific activities of glutamate and glutamine, but not of the other 3 amino acids. The preferred precursor for glutamate synthesis in native PC12 cells was glucose followed by octanoate, butyrate and palmitate (16:6:3:1).
Adrenal Gland Neoplasms↗
Mechanism of biosynthesis of unsaturated fatty acids in Pseudomonas sp. strain E-3, a psychrotrophic bacterium.
Biosynthesis of palmitic, palmitoleic, and cis-vaccenic acids in Pseudomonas sp. strain E-3 was investigated with in vitro and in vivo systems. [1-14C]palmitic acid was aerobically converted to palmitoleate and cis-vaccenate, and the radioactivities on their carboxyl carbons were 100 and 43%, respectively, of the total radioactivity in the fatty acids. Palmitoyl coenzyme A desaturase activity was found in the membrane fraction. [1-14C]stearic acid was converted to octadecenoate and C16 fatty acids. The octadecenoate contained oleate and cis-vaccenate, but only oleate was produced in the presence of cerulenin. [1-14C]lauric acid was aerobically converted to palmitate, palmitoleate, and cis-vaccenate. Under anaerobic conditions, palmitate (62%), palmitoleate (4%), and cis-vaccenate (34%) were produced from [1-14C]acetic acid, while they amounted to 48, 39, and 14%, respectively, under aerobic conditions. In these incorporation experiments, 3 to 19% of the added radioactivity was detected in released 14CO2, indicating that part of the added fatty acids were oxidatively decomposed. Partially purified fatty acid synthetase produced saturated and unsaturated fatty acids with chain lengths of C10 to C18. These results indicated that both aerobic and anaerobic mechanisms for the synthesis of unsaturated fatty acid are operating in this bacterium.
Acetates↗
Effects of CDPcholine and CDPethanolamine on the alterations in rat brain lipid metabolism induced by global ischemia.
The fast turnover pool of rat brain lipids was labeled by intracerebral injection of [3H]acetate. Cerebral ischemia for a duration of 5 min after decapitation caused a 2.2-fold increase in radioactivity in the free fatty acids and loss of more than 20% of the radioactivity from choline and ethanolamine glycerophospholipids. An intracerebral injection of 0.6 mumol each of cytidine diphosphocholine (CDPcholine) and cytidine diphosphoethanolamine (CDPethanolamine) prevented the loss of radioactivity from the glycerophospholipids and decreased the amount of radioactivity in the free fatty acids by 59% as compared with control values and 82% as compared with ischemia values. By GLC assays of the mass of the free fatty acids there was a threefold increase of free fatty acids in ischemic brains. Pretreatment of ischemic brains with CDPcholine and CDPethanolamine reduced the levels of unesterified fatty acids to 60% of the control values. Thus, a prior injection of cytidine nucleotides prevented the release of free fatty acids observed in ischemic brains.
Acetates↗
Galactolipid Synthesis in Vicia faba Leaves: IV. Site(s) of Fatty Acid Incorporation into the Major Glycerolipids.
The fatty acids of the major glycerolipids of Vicia faba leaves were analyzed immediately following (14)CO(2) feeding. The leaves were fractionated into chloroplast and cytoplasmic fractions and the location of radioactivity in the fatty acids determined. The results indicate that the major site of incorporation of fatty acids is in the phospholipids. Phosphatidylcholine contained the highest level of radioactivity in the cytoplasmic fraction, whereas phosphatidylglycerol contained radioactivity in both the chloroplast and cytoplasmic fractions. The galactolipids contained very little radioactivity in comparison, this radioactivity being confined to high speed centrifugal fractions believed to contain the envelopes of the chloroplast. Our results suggest that phosphatidylcholine is a major site of incorporation of fatty acids (mainly in oleic acid) in the cytoplasm, whereas phosphatidylglycerol is also a site of incorporation involving both oleic and palmitic acids, inside and outside the chloroplast.
Journal Article↗