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Plasma amino acids in reptiles after feeding protein or amino acids and after injecting amino acids.

Turtles (Pseudemys scripta elegans) and alligators (Alligator mississipiensis) were fed fish, casein, or free amino acid mixtures, or were injected with free amino acid mixtures, quantities in each case being equivalent to about 42 mmoles of amino acids/kg body weight. The experiments were designed to determine whether protein synthesis normally precedes catabolism of the amino acids absorbed from digesting food. Both animals disposed of free or food-derived amino acids more rapidly than could be accounted for by catabolism alone, but the transient increases in turtle plasma concentrations consisted mostly of essential amino acids, whereas the alligator plasma showed little increase in essential amino acids and considerable rises in four nonessential amino acids, glutamine, glutamic acid, glycine and alanine. Despite some species variation, these two species with low metabolic rates apparently utilized protein synthesis to dispose of amino acid mixtures prior to the necessarily slow process of catabolism for energy.

Administration, Oral

[On the phenolic acids of vegetables. II. Hydroxycinnamic acids and hydroxybenzoic acids of fruit and seed vegetables (author's transl)].

Fruits of solanaceae (tomatoes, eggplant, and sweet peppers) almost exclusively contain hydroxycinnamic acid derivatives with caffeic acid dominating. Fruits of cucurbitaceae (cucumbers, melons, pumpkins, and zucchini) are extraordinary, because they show very low concentrations of phenolic acids (up to 10 mg/kg) accumulated in the peels. Peas and broad beans have relatively small contents of phenolic acids too. Their husks show like beans considerable concentrations of hydroxycinnamic acid derivatives with dominating p-coumaric acid. In the group of hydroxybenzoic acids derivatives of salicylic, gentisic and vanillic acid could be determined frequently, but mostly as traces.

Caffeic Acids

Micelle and acid-soap formation of linoleic acid and 13-L-hydroperoxylinoleic acid being substrates of lipoxygenase-1.

Surface tension measurements of linoleic acid solutions in 0.1 M sodiumborate buffer pH 10 at 23 degrees C showed that at increasing the linoleic acid concentration a sharp transition from monomers to micelles occurs at 167 micrometer. At pH 9 and 8 formation of acid-soap dimers from monomers starts at 60 micrometer and 21 micrometer respectively. The concentration range at which only monomers exist is therefore markedly reduced. For 13-L-hydroperoxylinoleic acid at pH 10 acid-soap formation still takes place, starting at approx. 220 micrometer. The total lipid concentration at which acid-soap or micelle formation starts in mixtures of linoleic acid and 13-L-hydroperoxylinoleic acid has been determined in relation to the molar ratio of both acids.

Kinetics

Triple-helical polynucleotides. Mixed triplexes of the poly(uridylic acid)-poly(adenylic acid)-poly(uridylic acid) class.

By the techniques of interferon induction in primary rabbit kidney cells "superinduced" with metabolic inhibitors, ultraviolet absorbance-temperature profiles, sensitivity to pancreatic ribonuclease A, and sucrose velocity gradient ultracentrifugation, a number of reactions between double-helical RNA and single-stranded RNA or DNA homopolymers were investigated. The polymers involved in these studies were poly(adenylic acid), poly(uridylic acid), poly(ribothymidylic acid), poly(5-bromouridylic acid), poly(deoxythymidylic acid), poly(deoxyuridylic acid), poly(3-methyluridylic acid), poly(2'-O-methyluridylic acid), and poly(2'-azido-2'-deoxyuridylic acid). Two different reaction courses, both leading to the formation of triple helices, were noted: (1) poly(Ux)-poly(A) + poly(Uy) leads to poly(Ux)-poly(A)-poly(Uy) if the Tm of poly(Ux)-poly(A) was higher than the Tm of poly(Uy)-poly(A); (2) poly(Ux)-poly(A) + poly(Uy) leads to poly(Uy)-poly(A)-poly(Ux) if the Tm of poly(Ux)-poly(A) was lower than the Tm of poly(Uy)-poly(A). In these equations, the homopolymer written to the left of poly(A) implies Watson-Crick hydrogen bonding whereas the polymer to the right of poly(A) is involved in Hoogsteen hydrogen bonding.

Animals

Comparison of the conversion rates of alpha-linolenic acid (18:3(n - 3)) and stearidonic acid (18:4(n - 3)) to longer polyunsaturated fatty acids in rats.

The delta 6-desaturase reaction is regarded to be the rate-limiting step in the conversion of linoleic acid (18:2(n - 6)) to arachidonic acid (20:4(n - 6)). The same is probably also the case with the conversion of alpha-linolenic acid (18:3(n - 3)) to eicosapentaenoic acid (20:5(n - 3)). However, there are very few in vivo studies that directly compared the conversion rate between 18:3(n - 3) and stearidonic acid (18:4(n - 3)), which is the delta 6-desaturated product of 18:3(n - 3). We compared this rate by feeding rats on a lipid-free diet supplemented with lard (9%, w/w) and 18:3(n - 3) ethyl ester (1%) diet or on a diet containing lard (9%) and 18:4(n - 3) ethyl ester (1%). A lard (10%)-supplemented diet was used as the control diet. The fatty acid compositions of total phospholipids, triglycerides and free fatty acids of both liver and plasma were measured after 1 or 3 weeks on different diets. The molar ratio of 20:5(n - 3) of most lipid fractions was about 2-fold higher in rats fed the 18:4(n - 3)-supplemented diet than in rats fed the 18:3(n - 3)-supplemented diet. 18:4(n - 3) was found in the liver lipid fraction in only a very small amount, even in the 18:4(n - 3)-supplemented groups. Thus, desaturation at C-6 is suggested to be the rate-limiting step in the conversion of 18:3(n - 3) to 20:5(n - 3).

Animals

Accumulation of 3-hydroxyisobutyric acid, 2-methyl-3-hydroxybutyric acid and 3-hydroxyisovaleric acid in ketoacidosis.

1. Urine and serum samples from patients with ketoacidosis of varying degree and etiology have been examined by gas chromatography and mass spectrometry. 2. In addition to 3-hydroxyisovaleric acid, relatively high concentrations of two analogous hydroxy acids, 3-hydroxyisobutyric acid and 2-methyl-3-hydroxybutyric acid, were found in the urine. 3. There were highly significant positive correlations between the excreted amounts of the three acids. 4. Experiments on rats with isotope-labelled compounds revealed that the acids were formed by the degradation of leucine, isoleucine and valine. 5. The accumulation of the hydroxy acids during ketoacidosis is probably caused by a similar derangement of the metabolism of all three branched-chain amino acids.

Acetates

[Investigations on the utilisation of parenterally administered amino acids by premature and small-for-dates neonates. II. Investigations of elimination half life time, elimination constants, transfer and clearances of amino acids after short-time infusion of L-amino acid mixtures (author's transl)].

10 ml/kg body weight of a 5% standardized L-amino-acid-mixture was given within the first 25 days to 25 prematurely born infants of 32--37 weeks gestationale and 5 small-for-dates newborns. We studied the metabolic kinetics of these amino acids in the serum and calculated elimination half life time, elimination constants, clearance and transfer rates of the single amino acids. The results were correlated with sex, nutritional state and age. 5 additional experiments were performed with 3 prematurely born infants and 2 small-for-dates newborns. The following results were of special interests: 1. Nearly all amino-acids had half life times between 30 and 50 min. Ornithine and tryptophane were more (half life time: 70 and about 50 min, respectively) slowly eliminated, while arginine and glutamic acid had half life times of between 5 and 15 min. 2. There was no statistically significant differences between sexes. 3. Eutrophic prematures eliminated the amino acids much faster then the hypotrophic ones. 4. Hypotrophic premature infants showed slow elimination of nearly all amino acids compared with hypotrophic newborns. 5. The elimination of nearly all amino acids studied grew faster with increasing age. The most significant differences were registered between the 1st and 3rd weeks of life.

Age Factors

Bile acid metabolism in mammals: IX. Conversion of chenodeoxycholic acid to cholic acid by isolated perfused rat liver.

Current dogma of bile acid synthesis in mammals insists that hydroxylation of the ring structure at C-12 precedes side chain oxidation, and that chenodeoxycholic acid is not converted to cholic acid under normal conditions. This report concerns the conversion of chenodeoxycholic acid to cholic acid by isolated, perfused rat liver. Results indicate that isolated perfused rat liver has a definite, but limited, capacity for synthesis of cholic acid from chenodeoxycholic acid.

Animals

Lipid compositional manipulation in Acholeplasma laidlawii B. Effect of exogenous fatty acids on fatty acid composition and cell growth when endogenous fatty acid production is inhibited.

A variety of potential inhibitors of de novo fatty acid biosynthesis have been tested for activity in Acholeplasma laidlawii B. Two compounds, avidin and N,N-dimethyl-4-oxo-2trans-dodecenamide (CM-55), an antimicrobial fatty amide, strongly inhibit de novo biosynthesis without nonspecific toxic effects at moderate dosages. Avidin is the more potent inhibitor, abolishing de novo fatty acid synthesis and greatly reducing the chain elongation of exogenous fatty acids at level of 25 U/l. CM-55 gives complete inhibition of de novo biosynthesis only at low temperatures and inhibits exogenous fatty acid elongation to a variable extent. However, CM-55 is still a more potent antilipogenic agent in this organism than is the fungal antibiotic cerulenin. Cells cultured with avidin grow only when one or more exogenous medium- or long-chain fatty acids are added to the growth medium. The extent of cell growth under these conditions depends primarily on the physical properties of the exogenous fatty acid(s). In general, fatty acids giving diacylglycerolipids of very high or very low fluidity are unsuitable growth substrates, while those whose diacylglycerol derivatives are of intermediate fluidity support fair to good cell growth.

Acholeplasma laidlawii

Reinitiation of deoxyribonucleic acid synthesis by deoxyribonucleic acid initiation mutants of Escherichia coli: role of ribonucleic acid synthesis, protein synthesis, and cell division.

The dnaA and dnaC genes are thought to code for two proteins required for the initiation of chromosomal deoxyribonucleic acid replication in Escherichia coli. When a strain carrying a mutation in either of these genes is shifted from a permissive to a restrictive temperature, chromosome replication ceases after a period of residual synthesis. When the strains are reincubated at the permissive temperature, replication again resumes after a short lag. This reinitiation does not require either protein synthesis (as measured by resistance to chloramphenicol) or ribonucleic acid synthesis (as measured by resistance to rifampin). Thus, if there is a requirement for the synthesis of a specific ribonucleic acid to initiate deoxyribonucleic acid replication, this ribonucleic acid can be synthesized prior to the time of initiation and is relatively stable. Furthermore, the synthesis of this hypothetical ribonucleic acid does not require either the dnaA of dnaC gene products. The buildup at the restrictive temperature of the potential to reinitiate deoxyribonucleic acid synthesis at the permissive temperature shows rather complex kinetics the buildup roughly parallels the rate of mass increase of the culture for at least the first mass doubling at the restrictive temperature. At later times there appears to be a gradual loss of initiation potential despite a continued increase in mass. Under optimal conditions the increase in initiation potential can equal, but not exceed, the increase in cell division at the restrictive temperature. These results are most easily interpreted according to models that postulate a relationship between the initiation of deoxyribonucleic acid synthesis and the processes leading to cell division.

Bacterial Proteins

Faecal bile acid loss and bile acid pool size during short-term treatment with ursodeoxycholic and chenodeoxycholic acid in patients with radiolucent gallstones.

Twelve non-obese patients with radiolucent gallstones were fed on a standard diet. After 10 days (period A), six patients received 15 mg/kg/day of ursodeoxycholic acid (UDCA) (group I) and the other six (group II) the same dose of chenodeoxycholic acid (CDCA) for 15 days (period B). An intravenous injection of 20 micro Ci of 14C-UDCA and of 14C-CDCA was given on the 11th day of period B to the patients of group I and II respectively. Stools were collected at the end of period A and B and one bile sample was collected on the 12th day of period B. The faecal bile acid loss was higher during chenotherapy (36.12 mumol/kg/day) than during ursotherapy (23.94 mumol/kg/day), as was the proportion of lithocholic acid (73% vs 43%) in the faeces. Decay constant rate of faecal radioactivity was 0.365 day-1 in group I and 0.642 in group II. The results indicate that faecal bile acid excretion and turnover rate are greater during CDCA than UDCA, while UDCA increases the bile acid pool size to an even greater extent than does CDCA (150.2 vs 94.9 mumol/kg). This is probably because the former is more slowly degraded to poorly reabsorbable compounds. In fact, the bile saturation index was 0.66 in group I and 1.05 in group II, even though biliary CDCA in the latter had risen to 69.6%.

Bile Acids and Salts

Simultaneous determination by GC-MS-SIM of o-, m-, p-hydroxyphenylacetic acid, 3:4-dihydroxyphenylacetic acid and homovanillic acid in biological samples using a common selected ion.

A GC-MS-SIM method is described for the simultaneous determination of five acid metabolites, o-, m-, and p-hydroxyphenylacetic acid, 3:4-dihydroxyphenylacetic acid and homovanillic acid in biological samples using a common selected ion for all the acids. The TMS ethers of methyl or ethyl esters of these compounds have a common ion at m/e 179 which is used for their quantitation by selected ion monitoring. The molecular ion at m/e 238 of the three monohydroxy phenylacetic acids, which is also the M-30 ion for homovanillic acid, is also used for their quantitation. The GC conditions for their separation, the relative sensitivities and some advantages of TMS ethers over other derivatives are described. The application of this method to biological samples is illustrated by data on human urine and plasma.

3,4-Dihydroxyphenylacetic Acid

The estimation of 3,4-dihydroxyphenylacetic acid, homovanillic acid and homo-isovanillic acid in nervous tissue by gas-liquid chromatography and electron capture detection.

1 A gas chromatographic method using electron capture detection is described for the estimation of three acidic metabolites of dopamine, 4-hydroxy-3-methoxyphenylacetic acid (homovanillic acid, HVA), 3,4-dihydroxyphenylacetic acid (DOPAC) and 3-hydroxy-4-methoxyphenylacetic acid (homo-isovanillic acid, iso-HVA). The method is based on the formation of the trifluoroacetyl-hexafluoroisopropyl derivatives of the three acids. 2 The method has been applied to the estimation of DOPAC, HVA and iso-HVA in tissues from the central and peripheral nervous systems.

3,4-Dihydroxyphenylacetic Acid

Downregulation of saturated fatty acid and upregulation of unsaturated fatty acid by 13-cis-retinoic acid in human prostate cancer cells.

Retinoids play a major role in regulation of epithelial cell growth and cellular differentiation, but their mechanism(s) of action are still unclear. In the present study, we examined the effects of 13-cis-retinoic acid (13-cis-RA) on cytotoxicity, growth properties, morphology, neutral lipids, phospholipids and fatty acids in cultured human prostate cancer cell lines. The results of these experiments suggest that 13-cis-RA (10 microM) inhibits the DNA synthesis and nude mice tumorigenicity by 2- to 3-fold, compared to control. Electron microscopy revealed more differentiated phenotypes after 13-cis-RA treatment. There was a significant increase in phosphatidylcholine and decrease in sphingomyelin in 13-cis-RA treated cells compared to control. The saturated fatty acids significantly decreased whereas unsaturated fatty acids were increased after 13-cis-RA treatment in prostate cancer cells. This study demonstrates for the first time that retinoic acid mediated downregulation of saturated fatty acids and upregulation of unsaturated fatty acid in human prostate cancer cells.

Animals

[A possible essential metabolite of linoleic acid: lipoic acid, the universal coenzyme of alpha-keto acid oxidation].

The metabolism of [14C-U] linoleci acid (LI) and [14C-U] oleic acid were compared by injecting these fatty acids into growing rats and then homogenizing livers into a three phase system (chloroform/methanol/water). The radioactivities of these phases were equilibrated by extracting them X times with the same solvents. The lipid lower phase was discarded and the analysis was carried out on the evaporated hydroalcoholic upper phase. The residue was extracted again with methanol and hydrolyzed (HCl 6N). The acidic solution was evaporated, treated with HCl/methanol, extracted with chloroform and analysed by thin layer chromatography. One of the most radioactive intermediates detected after injecting LI was purified again and identified as lipoic acid, on the basis of: a. retention time in gas-liquid chromatography; b. Rf in thin layer chromatography; c. molecular weight as determined by mass spectrometry. Thus, the most important fate of essential fatty acids (except for their part in the prostaglandin synthesis and membrane formation) seems to be that of a precursor for this covalently bound alpha keto-acid dehydrogenation coenzyme--the link between lipid and carbohydrate metabolisms.

Animals

Regulation of the biosynthesis of aminoacyl-transfer ribonucleic acid synthetases and of transfer ribonucleic acid in Escherichia coli. VI. Mutants with increased levels of glutaminyl-transfer ribonucleic acid synthetase and of glutamine transfer ribonucleic acid.

Spontaneous revertants of a temperature-sensitive Escherichia coli strain bearing a thermolabile glutaminyl-transfer ribonucleic acid (tRNA) synthetase have been selected for growth at 45 degrees C. Among 10 revertants still containing the thermolabile enzyme, 2 interesting strains were found. One strain has a fivefold elevated level of the thermolabile glutaminyl-tRNA synthetase; the genetic locus, glnR, responsible for this effect maps at min 24, far from glnS, the structural gene of the enzyme. In the other strain the levels of tRNA Gln and several other tRNAs are twice as high as in the parental strain; the locus responsible, glnU, maps at min 59.5 on the E. coli map.

Amino Acyl-tRNA Synthetases