N-(5′-Phosphopyridoxyl)-4-aminobutyric acid: a stable bisubstrate adduct inhibitor of rat brain 4-aminobutyric acid aminotransferase

Summary N-(5-Phosphopyridoxyl)-4-aminobutyric acid, a stable adduct of pyridoxal phosphate and 4-aminobutyric acid, has been shown to be a potent inhibitor of rat brain 4-aminobutyric acid aminotransferase (GABA-T) with a K_i of 1.4 M.Acknowledgments. This work was supported in part by the United Parkinson Foundation, l'Association Canadienne l'Ataxie de Friedreich, and the Medical Research Council of Canada.     

Metabolism of 3β-hydroxycholest-5-en-26-oic acid in hamsters

Summary Metabolism of 26-hydroxycholesterol to 3-hydroxychol-5-en-24-oic acid and other C24-bile acids has been expected to occur by way of 3-hydroxycholest-5-en-26-oic acid in studies in vitro. 3-Hydroxycholest-5-en-26-oic acid was infused intravenously into bile fistula hamsters and the following C24-bile acids were identified: 3-hydroxychol-5-en-24-oic acid, lithocholic acid, chenodeoxycholic acid, and a small amount of cholic acid.     

Eicosapentaenoic acid in tissue lipids ofPieris brassicae

Summary Larvae of the cabbage white butterfly,Pieris brassicae, have a dietary requirement for linolenic acid (C183n3) and were found to accumulate two other members of the n-3 family, C203n3 and C205n3 (eicosapentaenoic acid) especially in testicular phospholipids. Arachidonic acid was observed in trace amounts only. During diapause the relative titer of eicosapentaenoic acid increased in testicular phospholipids to about 4.2% of the fatty acids. Eicosapentaenoic acid is a possible precursor of prostaglandins, suggesting that prostaglandins of the 3-series predominate in this insect.     

Tannic acid inhibits insulin-stimulated lipogenesis in rat adipose tissue and insulin receptor function in vitro

Tannins occur naturally in relatively abundant amounts in fruits, herbal medicines and common beverages. Thus an understanding of how these polyphenols affect peptide hormone action is of importance. We report here that tannic acid (a hydrolysable tannin) inhibits insulin-stimulated lipogenesis in rat adipose tissue in vitro, with an IC₅₀ estimated to be about 350 M. However, its monomer, gallic acid, did not show a similar inhibitory effect at concentrations up to 1 mM. The inhibition by tannic acid was less evident with higher concentrations of bovine serum albumin in the incubation buffer. This was attributed to the formation of a tannin-protein complex between bovine serum albumin and tannic acid. In a binding assay, it was observed that the specific binding of insulin to its receptor was not inhibited by tannic acid in the concentration range 0–200 M. However, insulin-stimulated autophosphorylation of the insulin receptor, and receptor-associated tyrosine kinase phosphorylation of RR-SRC peptide, were inhibited by tannic acid at concentrations as low as 25 M. Our data do not support the current speculation that tannins affect the activity of peptide hormones by binding to them. Therefore, our finding opens up a new perspective in the understanding of the mode of action of tannins on such hormones.     

Preferential hydrophobic interactions are responsible for a preference of D-amino acids in the aminoacylation of 5′-AMP with hydrophobic amino acids

We have studied the chemistry of aminoacyl AMP to model reactions at the 3 terminus of aminoacyl tRNA for the purpose of understanding the origin of protein synthesis. The present studies relate to the D, L preference in the esterification of 5-AMP. All N-acetyl amino acids we studied showed faster reaction of the D-isomer, with a generally decreasing preference for D-isomer as the hydrophobicity of the amino acid decreased. The -branched amino acids, Ile and Val, showed an extreme preference for D-isomer. Ac-Leu, the -branched amino acid, showed a slightly low D/L ratio relative to its hydrophobicity. The molecular basis for these preferences for D-isomer is understandable in the light of our previous studies and seems to be due to preferential hydrophobic interaction of the D-isomer with ademine. The preference for hydrophobic D-amino acids can be decreased by addition of an organic solvent to the reaction medium. Conversely, peptidylation with Ac-PhePhe shows a preference for the LL isomer over the DD isomer.     

Tetrapyrrole biosynthesis from 4,5-dioxovaleric acid inRhodopseudomonas spheroides

Summary Porphyrin biosynthesis from 4,5-dioxovaleric acid was studied in cell suspensions ofR. spheroides. The experiments show that 4,5-dioxovaleric acid is a far precursor of porphyrins through amino laevulinic acid formation in a transamination reaction involving also 1-alanine. It differs radically from the classical aminolaevulinic acid synthesis using glycine and succinyl CoA as substrates.Acknowledgments: The authors are indebted to Dr Rodolfo García for helpful discussions and criticisms. Special thanks are given to Dr Aldo Mitta and other members of the Comisión Nacional de la Energía Atómica for their continuous help and support and for using their facilities in order to synthetise DVA.     

The biochemistry of ancient DNA in bone

The amount of DNA in ancient bone was determined by ethidium bromide staining after the removal of the potent Taq inhibitor, fulvic acid. A complete decalcification and a perfusion protocol were used to recover DNA from bone. A variety of purification techniques including molecular sieve, hydroxyapatite binding and Magic preparations yielded DNA that spanned from 3.4g/g of bone to below detectable limits. Fulvic acid was shown to interfere with the quantification of DNA derived from ancient human skeletal material one hundred to over seven thousand years old. Scanning UV in the 300 to 230 nm range is a simple and sensitive technique for documenting fulvic acid contamination in ancient bone extracts.     

Failure of ascorbic acid to influence brain catecholamines in the guinea-pig

Summary Lack of dietary ascorbic acid lowered plasma levels of ascorbic acid but failed to change levels of brain norepinephrine or dopamine.Investigators adhered during the research described in this report to the policy set forth in the Guide for the Care and Use of Laboratory Animals by the Committee on Revision of the Guide for Laboratory Animal Resources, National Research Council.The opinions or assertions contained herein are the private views of the authors and are not to be construed as official or as reflecting the views of the Department of the Army or the Department of Defense.     

Metabolic pathways of δ-aminolaevulinic acid inRhodopseudomonas spheroides

Summary Incorporation of -aminolaevulinic acid 5¹⁴C and 4¹⁴C into the inosine monophosphate pool and into porphyrins, was studied in cell suspensions ofR. spheroides. The results contradict a direct incorporation of -aminolaevulinic acid into the purine ring of nucleotides through -dioxovaleric acid. It would suggest a nonspecific incorporation after degradation of -aminolaevulinic acid without a transamination as a first reaction.     

Mechanisms of lipid peroxidation: Iron catalyzed decomposition of fatty acid hydroperoxides as the basis of hydrocarbon evolution in vivo

Summary Ethane and pentane are evolved during peroxidation of tissue lipids in vivo and are believed to be derived from 3 and 6 fatty acids respectively. We present evidence supporting a scission mechanism as the means of forming these hydrocarbons during hydroperoxide decomposition. A fatty acid 6 hydroperoxide (methyl 13-hydroperoxy-6,9,11-octatrienoate), when incubated with a ferrous ion, yielded pentane as the major hydrocarbon (98%). Reaction with ferrous ion results in an alkoxy radical capable of undergoing scission to an aldehydic fatty acid and a hydrocarbon.We wish to thank Dr Ned Porter for both his generous donation of a supply of hydroperoxide and for his suggestions regarding possible reaction mechanisms. This research was supported by U.S.P.H.S. N.I.H. Grants ES00798, HL16264 and T32 GM07105.     

Changes in the sialic acid content of chick thymus and bursa of Fabricius during age-involution

Summary The sialic acid content of both thymus and bursa of Fabricius during their growth and involution phases in chick has been reported in this study. It is observed that the sialic acid concentration is very high in 1-week-old chickens. The concentration subsequently decreases to a significant level and rises again prior to the onset of involution. In the postinvolution period, a more or less minimal and constant level is maintained. The role of sialic acid in cellular activities of thymo-bursal system has been discussed.Supported by the University Grants Commission, Govt of India under the scheme Support to Research.     

Wound healing properties of naphthaquinone pigments fromAlkanna tinctoria

Summary From the roots ofAlkanna tinctoria Tausch, the alkannin esters of the following acids were isolated and identified: ,-dimethylacrylic acid, -acetoxy-isovaleric acid, isovaleric acid, and angelic acid. These esteric pigments showed excellent wound healing properties in a clinical study on 72 patients with ulcus cruris.I thank Dr A. Winkler, Heidberg Hospital and Prof. K. Kanitakis, Aristotle University, for the clinical studies. Prof. A. Sagredos, Aristotle University, for his help and Dr C. Gairola, Tobacco and Health Research Institute, for the mutagenicity tests. I also thank Dr G. A. Digenis and Dr L. Hurley, University of Kentucky, College of Pharmacy, for their critical reviews and comments.     

Chelate aus Diarylborinsäuren undβ-Ketosäureestern

Summary Diarylborinic acids form stable chelates with-keto acid esters. Borinic acids, when being synthetized, can be stabilized in compounds resulting from the combination with-keto acid esters. The chelates so formed are suitable—like the free borinic acids or their anhydrides—for further transformation. These chelates have several advantages in comparison with those formed with-amino ethanol.     

Variation in the fatty acid composition of developing seeds of rapeseed

Summary A study was made of the variation in the fatty acid composition of the oil in the developing seed of the Rafal cultivar of theBrassica napus L. cultivated in central Italy. The oil content reaches its maximum level 60 days after the petals fall. The increase in the percentage of oleic acid is negatively correlated with the palmitic, stearic and linoleic acid content.     

Chemical characterization of human urine albumin in proteinuria

Summary From the urine of a patient with proteinuria, the albumin protein component was isolated and compared with human serum albumin. By comparing the amino acid composition of the original proteins and their large cyanogen bromide fragments, peptide maps and N-terminal sequences of 33 amino acid residues, the identity of both proteins was shown.We wish to express our thanks to Mr L. Slepika and Dr A. Koent for the protein fractionation. We are indebted to Mr K. Grüner for the Edman degradation experiments, to Mrs E. Drková for amino acid analyses and Mrs A. Kulhánková for technical assistance.     

Über einen möglichen Weg der Biosynthese der Carotinoide beiMucor hiemalis

Summary Our experiments up to now lead us to the hypothesis, that the biosynthesis of carotenoids may be started on acetyl-coenzyme A. The condensation of two molecules of acetyl-coenzyme A leads to acetoacetyl-coenzyme A which can be regarded as the precursor of-methylcrotonic acid. For a long time the latter compound was postulated as the precursor of the natural isoprene derivatives. The participation of the citric acid cycle in the biosynthesis of carotenoids is discussed.     

Inhibition of rat liver glutamine synthetase by phosphonic analogues of glutamic acid

Summary Analogues of glutamic acid, -methylglutamic acid and glutamine in which the -or -COOH groups are replaced by PO₃H₂ or P(O)(OH₃)OH functions competitively inhibit rat liver glutamine synthetase. The K₁ values are comparable to or lower than K_M for L-glutamate.This study was supported by grant R.1.9.     

Gamma-aminobutyric acid uptake by rat kidney brush-border membrane vesicles

Summary Brush-border membrane vesicles (BBMV) from rat kidney cortex possessed two uptake systems for -aminobutyric acid (GABA), a high affinity system (Km=10.9 M) and a low affinity system (Km=1203 M). Both uptake systems were inhibited by p-hydroxymercuribenzoic acid and ouabain, and by the action of neuraminidase, whereas the GABA analogs nipecotic acid, -alanine, 2,4-diaminobutyric acid and 4,5,6,7-tetrahydroisoxazolo-[4,5c]-pyridin-3-ol had no effect on the GABA uptake activity. The BBMW uptake systems were clearly different from the GABA transport systems present in brain tissue.     

Asscorbic acid portects against male infertility in a teleost fish

An animal unable to synthesize ascorbic acid uniquely minicks human and non-human primates. Therefore, in this study we used the rainbow trout, a teleost fish, as the model animal to study the importance of dietary ascorbic acid on the fertilizing ability of sperm. A high concentration of ascorbic acid in semen plays a key role in maintaining the genetic integrity of sperm cells, by preventing oxidative damage to sperm DNA. This study will show that the concentration of asorbic acid in seminal plasma refelcts the dietary fed either an ascorbate-free diet (from 4.74±0.9 to 0.16±0.08 g ml^–1) or an ascorbate-rich diet (from 37.9±4.7 to 17.7± 3.2 g ml^–1) during the sperimnation season. The relationship between ascrobate status and fertility was studied in six groups of fish fed graded levels of ascorbic acid, which sperimated over a 150-day-period. Sperm from individual males was used to fertilize several batches of eggs. When the seminal plasma ascorbate concentration decreased to 7.3 g ml^–1 a significant decrease of fertilization rate and the hatching rate of embryos resulted. This is the first evidence that dietary ascorbate level directly affected sperm quality and influenced male fertility in a scruvy-prone vertebrate.     

The oxidative metabolism ofα-chlorohydrin and the chemical induction of spermatocoeles

Summary -Chlorohydrin (I) is oxidatively metabolized to -chlorolactic acid (III) and oxalic acid (IV). Deposition of calcium oxalate within the renal tubules is responsible for the toxic effects of-chlorohydrin and a similar action on the epididymis or epididymal blood vessels could initiate the formation of spermatocoeles from this and other male antifertility agents.