Synthesis and GGCT Inhibitory Activity of N-Glutaryl-L-alanine Analogues.

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Ii, Hiromi; Yoshiki, Tatsuhiro; Hoshiya, Naoyuki; Uenishi, Jun'ichi

2016-01-01

γ-Glutamylcyclotransferase (GGCT) is an important enzyme that cleaves γ-glutamyl-amino acid in the γ-glutamyl cycle to release 5-oxoproline and amino acid. Eighteen N-acyl-L-alanine analogues including eleven new compounds have been synthesized and examined for their inhibitory activity against recombinant human GGCT protein. Simple N-glutaryl-L-alanine was found to be the most potent inhibitor for GGCT. Other N-glutaryl-L-alanine analogues having methyl and dimethyl substituents at the 2-position were moderately effective, while N-(3R-aminoglutary)-L-alanine, the substrate having an (R)-amino group at the 3-position or N-(N-methyl-3-azaglutaryl)-L-alanine, the substrate having an N-methyl substituent on the 3-azaglutaryl carbon, in constract, exhibited excellent inhibition properties.

The concentration of free amino acids in blood serum of dairy cows with primary ketosis.

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Marczuk, J; Brodzki, P; Brodzki, A; Kurek, Ł

2018-03-01

Ketosis is a common condition found in the initial stages of lactation in high-yielding dairy cows. The major cause of ketosis is a negative energy balance. During the energy deficiency, proteolysis processes develop parallel to lipolysis. During proteolysis, muscle tissue can be used as a source of amino acid. To date, the participation of amino acids in gluconeogenesis (glucogenic amino acids) and ketogenesis (ketogenic amino acids) has not been determined in detail. This paper presents the study on determination of the parameters of protein and free amino acid metabolism in blood serum of dairy cows with primary ketosis compared to healthy cows. This study contributes to better understanding of the role of amino acids in pathogenesis of ketosis. A total of 30 cows, divided into two groups: experimental (15 cows with ketosis) and control (15 healthy cows), were included in the study. The concentrations of glucose, β-hydroxybutyrate, total protein, albumin, urea, and free amino acids were determined in peripheral blood. Statistically significantly higher concentrations of glutamine, glutamic acid, isoleucine (p≤0.001), and tyrosine (p≤0.05) were found in cows with primary ketosis compared to healthy cows. Significant decrease in the concentrations of asparagine, histidine, methionine, and serine (p≤0.001), alanine, leucine, lysine and proline (p≤0.05) was observed. Significant increase of total ketogenic and glucogenic amino acids (p≤0.05), and an increased ratio of total ketogenic and glucogenic amino acids to total amino acids (p≤0.001) were noted in cows with ketosis. In our study, the changes, in particular observed in amino acid concentration in cows with primary ketosis, indicate its intensive use in both ketogenesis and gluconeogenesis processes. Therefore, a detailed understanding of the role that amino acids play in gluconeogenesis and ketogenesis will improve ketosis diagnostics and monitoring the course of a ketosis episode. Perhaps, the

Alterations in serum amino acid concentrations in dogs with protein-losing enteropathy.

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Kathrani, Aarti; Allenspach, Karin; Fascetti, Andrea J; Larsen, Jennifer A; Hall, Edward J

2018-03-31

Certain amino acids are decreased in humans with inflammatory bowel disease (IBD) and supplementation with the same amino acids has shown beneficial effects in animal models of IBD. Currently, the amino acid status of dogs with protein-losing enteropathy (PLE) is unknown. To determine if serum amino acid concentrations are abnormal in dogs with PLE and correlated with clinical and laboratory variables and outcome. Thirty client-owned dogs diagnosed with PLE and 12 apparently healthy dogs seen at Bristol Veterinary School. Retrospective study using stored residual serum from fasted dogs with PLE, collected at the time of diagnostic investigation and from apparently healthy dogs. Serum was analyzed for 30 amino acids using an automated high-performance liquid chromatography amino acid analyzer. Serum tryptophan concentrations were significantly decreased in dogs with PLE (median, 22 nmol/mL; range, 1-80 nmol/mL) compared with apparently healthy control dogs (median, 77.5 nmol/mL; range, 42-135 nmol/mL, P PLE and apparently healthy. Serum tryptophan concentrations were also significantly correlated with serum albumin concentrations in dogs with PLE (P = .001, R 2 = 0.506). Decreased serum tryptophan concentration might play a role in the pathogenesis of canine PLE or be a consequence of the disease. © 2018 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals, Inc. on behalf of the American College of Veterinary Internal Medicine.

Neurotoxic Non-proteinogenic Amino Acid β-N-Methylamino-L-alanine and Its Role in Biological Systems.

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Popova, A A; Koksharova, O A

2016-08-01

Secondary metabolites of photoautotrophic organisms have attracted considerable interest in recent years. In particular, molecules of non-proteinogenic amino acids participating in various physiological processes and capable of producing adverse ecological effects have been actively investigated. For example, the non-proteinogenic amino acid β-N-methylamino-L-alanine (BMAA) is neurotoxic to animals including humans. It is known that BMAA accumulation via the food chain can lead to development of neurodegenerative diseases in humans such as Alzheimer's and Parkinson's diseases as well as amyotrophic lateral sclerosis. Moreover, BMAA can be mistakenly incorporated into a protein molecule instead of serine. Natural sources of BMAA and methods for its detection are discussed in this review, as well as the role of BMAA in metabolism of its producers and possible mechanisms of toxicity of this amino acid in different living organisms.

Solved? The reductive radiation chemistry of alanine.

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Pauwels, Ewald; De Cooman, Hendrik; Waroquier, Michel; Hole, Eli O; Sagstuen, Einar

2014-02-14

The structural changes throughout the entire reductive radiation-induced pathway of l-α-alanine are solved on an atomistic level with the aid of periodic DFT and nudged elastic band (NEB) simulations. This yields unprecedented information on the conformational changes taking place, including the protonation state of the carboxyl group in the "unstable" and "stable" alanine radicals and the internal transformation converting these two radical variants at temperatures above 220 K. The structures of all stable radicals were verified by calculating EPR properties and comparing those with experimental data. The variation of the energy throughout the full radiochemical process provides crucial insight into the reason why these structural changes and rearrangements occur. Starting from electron capture, the excess electron quickly localizes on the carbon of a carboxyl group, which pyramidalizes and receives a proton from the amino group of a neighboring alanine molecule, forming a first stable radical species (up to 150 K). In the temperature interval 150-220 K, this radical deaminates and deprotonates at the carboxyl group, the detached amino group undergoes inversion and its methyl group sustains an internal rotation. This yields the so-called "unstable alanine radical". Above 220 K, triggered by the attachment of an additional proton on the detached amino group, the radical then undergoes an internal rotation in the reverse direction, giving rise to the "stable alanine radical", which is the final stage in the reductive radiation-induced decay of alanine.

Compound-specific nitrogen isotope analysis of D-alanine, L-alanine, and valine: application of diastereomer separation to delta15N and microbial peptidoglycan studies.

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Takano, Yoshinori; Chikaraishi, Yoshito; Ogawa, Nanako O; Kitazato, Hiroshi; Ohkouchi, Naohiko

2009-01-01

We have developed an analytical method to determine the compound-specific nitrogen isotope compositions of individual amino acid enantiomers using gas chromatography/combustion/isotope ratio mass spectrometry. A novel derivatization of amino acid diastereomers by optically active (R)-(-)-2-butanol or (S)-(+)-2-butanol offers two advantages for nitrogen isotope analysis. First, chromatographic chiral separation can be achieved without the use of chiral stationary-phase columns. Second, the elution order of these compounds on the chromatogram can be switched by a designated esterification reaction. We applied the method to the compound-specific nitrogen isotope analysis of D- and L-alanine in a peptidoglycan derived from the cell walls of cultured bacteria (Firmicutes and Actinobacteria; Enterococcus faecalis, Staphylococcus aureus, Staphylococcus staphylolyticus, Lactobacillus acidophilus, Bacillus subtilis, Micrococcus luteus, and Streptomyces sp.), natural whole bacterial cells (Bacillus subtilis var. natto), (pseudo)-peptidoglycan from archaea (Methanobacterium sp.), and cell wall from eukaryota (Saccharomyces cerevisiae). We observed statistically significant differences in nitrogen isotopic compositions; e.g., delta15N ( per thousand vs air) in Staphylococcus staphylolyticus for d-alanine (19.2 +/- 0.5 per thousand, n = 4) and L-alanine (21.3 +/- 0.8 per thousand, n = 4) and in Bacillus subtilis for D-alanine (6.2 +/- 0.2 per thousand, n = 3) and L-alanine (8.2 +/- 0.4 per thousand, n = 3). These results suggest that enzymatic reaction pathways, including the alanine racemase reaction, produce a nitrogen isotopic difference in amino acid enantiomers, resulting in 15N-depleted D-alanine. This method is expected to facilitate compound-specific nitrogen isotope studies of amino acid stereoisomers.

Excitatory amino acid b-N-methylamino-L-alanine is a putative environmental neurotoxin

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VLADIMIR NEDELJKOV

2011-04-01

Full Text Available The amino acid b-N-methylamino-L-alanine (L-BMAA has been associated with the amyotrophic lateral sclerosis/parkinsonism-dementia complex in three distinct western Pacific populations. The putative neurotoxin is produced by cyanobacteria, which live symbiotically in the roots of cycad trees. L-BMAA was thought to be a threat only to those few populations whose diet and medicines rely heavily on cycad seeds. However, the recent discovery that cyanobacteria from diverse terrestrial, freshwater, and saltwater ecosystems around the world produce the toxin requires a reassessment of whether it poses a larger health threat. Therefore, it is proposed that monitoring L-BMAA levels in cyanobacteria-contaminated water supplies might be prudent.

Excess of L-Alanine in Amino Acids Synthesized in a Plasma Torch Generated by a Hypervelocity Meteorite Impact Reproduced in the Laboratory

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Managadze, George G.; Engle, Michael H.; Getty, Stephanie A.; Wurz, Peter; Brinckerhoff, William B.; Shokolov, Anatoly; Sholin, Gennady; Terent'ev, Sergey A.; Chumikov, Alexander E.; Skalkin, Alexander S

2016-01-01

We present a laboratory reproduction of hypervelocity impacts of a carbon containing meteorite on a mineral substance representative of planetary surfaces. The physical conditions of the resulting impact plasma torch provide favorable conditions for abiogenic synthesis of protein amino acids: We identified glycine and alanine, and in smaller quantities serine, in the produced material. Moreover, we observe breaking of alanine mirror symmetry with L excess, which coincides with the bioorganic world. Therefore the selection of L-amino acids for the formation of proteins for living matter could have been the result from plasma processes occurring during the impact meteorites on the surface. This indicates that the plasma torch from meteorite impacts could play an important role in the formation of biomolecular homochirality. Thus, meteorite impacts possibly were the initial stage of this process and promoted conditions for the emergence of a living matter.

Metabolism of [15N]alanine in the ectomycorrhizal fungus Paxillus involutus

International Nuclear Information System (INIS)

Chalot, M.; Finlay, R.D.; Ek, H.; Söderström, B.

1995-01-01

Chalot, M., Finlay, R. D., Ek, H., and Söderström, B. 1995. Metabolism of [ 15 N]alanine in the ectomycorrhizal fungus Paxillus involutus. Experimental Mycology 19, 297-304. Alanine metabolism in the ectomycorrhizal fungus Paxillus involutus was investigated using [ 15 N]alanine. Short-term exposure of mycelial discs to [ 15 N]alanine showed that the greatest flow of 15 N was to glutamate and to aspartate. Levels of enrichment were as high as 15-20% for glutamate and 13-18% for aspartate, whereas that of alanine reached 30%. Label was also detected in the amino-N of glutamine and in serine and glycine, although at lower levels. Preincubation of mycelia with aminooxyacetate, an inhibitor of transamination reactions. resulted in complete inhibition of the flow of the label to glutamate, aspartate, and amino-N of glutamine, whereas [ 15 N]alanine rapidly accumulated. This evidence indicates the direct involvement of alanine aminotransferase for translocation of 15 N from alanine to glutamate. Alanine may be a convenient reservoir of both nitrogen and carbon. (author)

Free amino acids in fibromyalgia syndrome: relationship with clinical picture.

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Ruggiero, Valeria; Mura, Massimiliano; Cacace, Enrico; Era, Benedetta; Peri, Marcella; Sanna, Giuseppina; Fais, Antonella

2017-04-01

The objectives of our study were to evaluate free amino acid (FAA) concentrations in the serum of patients affected by fibromyalgia syndrome (FMS) and to determine the relationships between FAA levels and FMS clinical parameters. Thus, serum amino acid concentrations were quantified (HPLC analysis) in 23 females with fibromyalgia (according to the American College of Rheumatology classification criteria) and 20 healthy females. The results showed significantly higher serum concentrations of aspartate, cysteine, glutamate, glycine, isoleucine, leucine, methionine, ornithine, phenylalanine, sarcosine, serine, taurine, tyrosine and valine in FMS patients vs. healthy controls. Patients with higher Fibromyalgia Impact Questionnaire (FIQ) scores showed increased levels of alanine, glutamine, isoleucine, leucine, phenylalanine, proline and valine. In conclusion, our results indicate an imbalance in some FAAs in FMS patients. Increased Glu is particularly interesting, as it could explain the deficit in monoaminergic transmission involved in pain.

Sulfur amino acids and alanine on pyrite (100) by X-ray photoemission spectroscopy: Surface or molecular role?

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Sanchez-Arenillas, M.; Galvez-Martinez, S.; Mateo-Marti, E.

2017-08-01

This paper describes the first successful adsorption of the cysteine, cystine, methionine and alanine amino acids on the pyrite (100) surface under ultra-high vacuum conditions with crucial chemical adsorption parameters driving the process. We have demonstrated by X-ray photoemission spectroscopy (XPS) that the surface pretreatment annealing process on pyrite surfaces is a critical parameter driving surface reactivity. The presence of enriched monosulfide species on the pyrite (100) surface favours the amino acid NH2 chemical form, whereas a longer annealing surface pretreatment of over 3 h repairs the sulfur vacancies in the pyrite, enriching disulfide species on the pyrite surface, which promotes NH3+ adsorption due to the sulfur vacancies in the pyrite being replaced by sulfur atom dimers (S22-) on the surface. Furthermore, even if the surface chemistry (monosulfide or disulfide species enrichment) is the main factor promoting a partial conversion from NH2 to NH3+ species, the unique chemical structure of each amino acid provides a particular fingerprint in the process.

Quantum mechanical effects in zwitterionic amino acids: The case of proline, hydroxyproline, and alanine in water

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Ulman, Kanchan; Busch, Sebastian; Hassanali, Ali A.

2018-06-01

In this work, we use ab initio molecular dynamics simulations to elucidate the electronic properties of three hydrated zwitterionic amino acids, namely proline, hydroxyproline, and alanine, the former two forming an important constituent of collagen. In all three systems, we find a substantial amount of charge transfer between the amino acids and surrounding solvent, which, rather surprisingly, also involves the reorganization of electron density near the hydrophobic non-polar groups. Water around proline appears to be slightly more polarized, as reflected by the enhanced water dipole moment in its hydration shell. This observation is also complemented by an examination of the IR spectra of the three systems where there is a subtle red and blue shift in the O-H stretch and bend regions, respectively, for proline. We show that polarizability of these amino acids as revealed by a dipole moment analysis involves a significant enhancement from the solvent and that this also involves non-polar groups. Our results suggest that quantum mechanical effects are likely to be important in understanding the coupling between biomolecules and water in general and in hydrophobic interactions.

Potential Role of Amino Acid/Protein Nutrition and Exercise in Serum Albumin Redox State

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Yasuaki Wada

2017-12-01

Full Text Available Albumin is the major protein in the serum of mammals. It is synthesized exclusively in the liver, before being secreted into the circulation. Similar to skeletal muscle protein, albumin synthesis is stimulated by dietary amino acids and proteins as well as exercise. Albumin has three isoforms based on the redox states of the free cysteine residue at position 34. The redox state of serum albumin has long been extensively investigated in terms of oxidative stress-related chronic diseases, with the redox state of serum albumin having been regarded as a marker of systemic oxidative stress. However, according to recent animal studies, the redox state of serum albumin is modulated by albumin turnover and may also reflect amino acid/protein nutritional status. Furthermore, as the redox state of serum albumin is modulated by exercise training, measuring the pre- and post-exercise redox states of serum albumin in athletes may be useful in assessing amino acid/protein nutritional status and exercise-induced oxidative stress, which are closely associated with skeletal muscle adaptive responses. This article extensively reviews serum albumin and the redox state of albumin in the context of amino acid/protein nutritional status and exercise training.

Blood metabolomics analysis identifies abnormalities in the citric acid cycle, urea cycle, and amino acid metabolism in bipolar disorder.

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Yoshimi, Noriko; Futamura, Takashi; Kakumoto, Keiji; Salehi, Alireza M; Sellgren, Carl M; Holmén-Larsson, Jessica; Jakobsson, Joel; Pålsson, Erik; Landén, Mikael; Hashimoto, Kenji

2016-06-01

Bipolar disorder (BD) is a severe and debilitating psychiatric disorder. However, the precise biological basis remains unknown, hampering the search for novel biomarkers. We performed a metabolomics analysis to discover novel peripheral biomarkers for BD. We quantified serum levels of 116 metabolites in mood-stabilized male BD patients (n = 54) and age-matched male healthy controls (n = 39). After multivariate logistic regression, serum levels of pyruvate, N-acetylglutamic acid, α-ketoglutarate, and arginine were significantly higher in BD patients than in healthy controls. Conversely, serum levels of β-alanine, and serine were significantly lower in BD patients than in healthy controls. Chronic (4-weeks) administration of lithium or valproic acid to adult male rats did not alter serum levels of pyruvate, N-acetylglutamic acid, β-alanine, serine, or arginine, but lithium administration significantly increased serum levels of α-ketoglutarate. The metabolomics analysis demonstrated altered serum levels of pyruvate, N-acetylglutamic acid, β-alanine, serine, and arginine in BD patients. The present findings suggest that abnormalities in the citric acid cycle, urea cycle, and amino acid metabolism play a role in the pathogenesis of BD.

Alanine scan of the peptide antibiotic feglymycin: assessment of amino acid side chains contributing to antimicrobial activity.

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Hänchen, Anne; Rausch, Saskia; Landmann, Benjamin; Toti, Luigi; Nusser, Antje; Süssmuth, Roderich D

2013-03-18

The antibiotic feglymycin is a linear 13-mer peptide synthesized by the bacterium Streptomyces sp. DSM 11171. It mainly consists of the nonproteinogenic amino acids 4-hydroxyphenylglycine and 3,5-dihydroxyphenylglycine. An alanine scan of feglymycin was performed by solution-phase peptide synthesis in order to assess the significance of individual amino acid side chains for biological activity. Hence, 13 peptides were synthesized from di- and tripeptide building blocks, and subsequently tested for antibacterial activity against Staphylococcus aureus strains. Furthermore we tested the inhibition of peptidoglycan biosynthesis enzymes MurA and MurC, which are inhibited by feglymycin. Whereas the antibacterial activity is significantly based on the three amino acids D-Hpg1, L-Hpg5, and L-Phe12, the inhibitory activity against MurA and MurC depends mainly on L-Asp13. The difference in the position dependence for antibacterial activity and enzyme inhibition suggests multiple molecular targets in the modes of action of feglymycin. Copyright © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Trend analysis of the correlation of amino acid plasma profile with glycemic status in Saudi diabetic patients

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Fahad A. Al-Abbasi

2012-10-01

Full Text Available The role of amino acids in diabetes mellitus and its metabolic traits have been suggested previously; however, studied to a very limited scale in the Saudi patient population. Patients diagnosed with diabetes mellitus were included in the current clinical study. Sample was representative and in accordance with the national population distribution. Blood samples were drawn and assayed for glucose, total cholesterol, triglyceride, high density lipoprotein and low density lipoprotein. General biochemical markers, such as alkaline phosphatase (ALP, creatinine kinase (CK, aspartate transaminase (AST, alanine transaminase (ALT and blood urea nitrogen (BUN were assessed. Serum amino acids of different categories (essential, semi-essential and metabolic indicator amino acids were assessed. Correlation co-efficient between each amino acid and serum glucose level was calculated. The current study showed positive correlation between amino acid level and glucose serum concentration in male while it showed negative correlation in female Saudi diabetic patients. Male patients had significantly higher methionine concentration parallel to their glycemic status. Metabolic indicator amino acids significantly changed in concordance with the glycemic status of female patients more than in male patients. In conclusion, serum amino acid is positively correlated with glycemic status in Saudi male diabetic patients while negatively correlated in female patients. Yet, further study would be recommended to utilize serum amino acid profile as surrogate parameter for the metabolic complications of diabetes mellitus.

Purification and characterization of l,(l/d)-aminopeptidase from Guinea pig serum.

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Krstanović, Marina; Brgles, Marija; Halassy, Beata; Frkanec, Ruza; Vrdoljak, Anto; Branović, Karmen; Tomasić, Jelka; Benedetti, Fabio

2006-01-01

Mammalian sera contain enzymes that catalyze the hydrolytic degradation of peptidoglycans and molecules of related structure and are relevant for the metabolism of peptidoglycans. We now report on a novel L,(L/D)-aminopeptidase found in human and mammalian sera. The enzyme hydrolyses the pentapeptide L-Ala-D-iso-Gln-meso-DAP(omegaNH(2))-D-Ala-D-Ala yielding the free L-alanine and the respective tetrapeptide (K(M) 18 mM). L,(L/D)-aminopeptidase from guinea pig serum was highly purified in four chromatographic steps, up to 700-fold. Molecular weight of the enzyme was estimated by HPLC to be approximately 175,000. The configuration of alanine obtained by hydrolysis of the pentapeptide was determined by oxidation with L-amino acid oxidase. The amino acids sequence in the respective tetrapeptide was deduced from the results of mass spectrometry. The novel L,(L/D)-aminopeptidase also hydrolyzed alanine-4-nitroanilide (K(M)=0.6 mM) and several peptides comprising L-amino acids. Peptides containing D-amino acid at the amino end and L-Asp-L-Asp were not the substrates for this enzyme. The purified enzyme also exhibited enkephalin degrading activity, hydrolyzing enkephalins comprising L,L- and L,D-peptide bonds. The enzyme was inhibited strongly by metal chelating agents, bestatin and amastatin.

Effect of sodium caproate on the volumetric and viscometric properties of glycine, DL-α-alanine, and DL-α-amino-n-butyric acid in aqueous solutions

International Nuclear Information System (INIS)

Wang Jianji; Yan Zhenning; Lu Jinsuo

2004-01-01

The apparent molar volumes (V m,2 ) and relative viscosities (η r ) at T=(298.15 and 308.15) K have been obtained for glycine, DL-α-alanine, and DL-α-amino-butyric acid in aqueous sodium caproate solutions from measurements of density and the flow time. The standard partial molar volumes (V 0 m,2 ), standard volumes of transfer (Δ t V 0 ), the viscosity B-coefficients, and the activation thermodynamic quantities (Δμ 2 0≠ and ΔS 2 0≠ ) of viscous flow have been calculated for the amino acids. It is shown that the standard partial molar volumes, viscosity B-coefficients, and activation free energies for viscous flow increase with increasing number of carbon atoms in the alkyl chain of the amino acids. An increase in V 0 m,2 and Δ t V 0 with increasing electrolyte concentrations have been explained due to the interactions of sodium caproate with the charged center of zwitterions for the amino acids. A comparison of the V 0 m,2 values for glycine, DL-α-alanine, and DL-α-aminon-n-butyric acid in different aqueous salts solutions showed that carboxylate ions have stronger interactions with amino acid than chloride, thiocyanate, and nitrate ions. Results of viscosity are discussed in terms of changes in solvent structure

Imaging analysis of direct alanine uptake by rice seedlings

International Nuclear Information System (INIS)

Nihei, Naoto; Masuda, Sayaka; Rai, Hiroki; Nakanishi, Tomoko M.

2008-01-01

We presented alanine, a kind of amino acids, uptake by a rice seedling to study the basic mechanism of the organic fertilizer effectiveness in organic farming. The rice grown in the culture solution containing alanine as a nitrogen source absorbed alanine approximately two times faster than that grown with NH 4 + from analysis of 14 C-alanine images by Imaging Plate method. It was suggested that the active transport ability of the rice seeding was induced in roots by existence of alanine in the rhizosphere. The alanine uptake images of the rice roots were acquired every 5 minutes successively by the real-time autoradiography system we developed. The analysis of the successive images showed that alanine uptake was not uniform throughout the root but especially active at the root tip. (author)

Secretion of d-alanine by Escherichia coli.

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Katsube, Satoshi; Sato, Kazuki; Ando, Tasuke; Isogai, Emiko; Yoneyama, Hiroshi

2016-07-01

Escherichia coli has an l-alanine export system that protects the cells from toxic accumulation of intracellular l-alanine in the presence of l-alanyl-l-alanine (l-Ala-l-Ala). When a DadA-deficient strain was incubated with 6.0 mM l-Ala-l-Ala, we detected l-alanine and d-alanine using high-performance liquid chromatography (HPLC) analysis at a level of 7.0 mM and 3.0 mM, respectively, after 48 h incubation. Treatment of the culture supernatant with d-amino acid oxidase resulted in the disappearance of a signal corresponding to d-alanine. Additionally, the culture supernatant enabled a d-alanine auxotroph to grow without d-alanine supplementation, confirming that the signal detected by HPLC was authentic d-alanine. Upon introduction of an expression vector harbouring the alanine racemase genes, alr or dadX, the extracellular level of d-alanine increased to 11.5 mM and 8.5 mM, respectively, under similar conditions, suggesting that increased metabolic flow from l-alanine to d-alanine enhanced d-alanine secretion. When high-density DadA-deficient cells preloaded with l-Ala-l-Ala were treated with 20 µM carbonyl cyanide m-chlorophenyl hydrazone (CCCP), secretion of both l-alanine and d-alanine was enhanced ~twofold compared with that in cells without CCCP treatment. In contrast, the ATPase inhibitor dicyclohexylcarbodiimide did not exert such an effect on the l-alanine and d-alanine secretion. Furthermore, inverted membrane vesicles prepared from DadA-deficient cells lacking the l-alanine exporter AlaE accumulated [3H]D-alanine in an energy-dependent manner. This energy-dependent accumulation of [3H]D-alanine was strongly inhibited by CCCP. These results indicate that E. coli has a transport system(s) that exports d-alanine and that this function is most likely modulated by proton electrochemical potential.

The fluctuation of free amino acids in serum during acute ischemic stroke

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Szpetnar Maria

2016-12-01

Full Text Available Currently, little data exists regarding the involvement of free amino acids (AA in the pathogenesis of ischemic stroke (IS. Thus, our objective was to study the degree of the degree of fluctuation of free amino acids level in serum during the acute phase of IS. The study consisted of eighteen patients (female/male: 10/8; age: 73.1 ± 4.1 with acute IS that was confirmed by way of computed tomography, while twelve sex and age matched individuals were assigned as control group. During the study period, the patients did not receive any supplemental amino acids therapy that could affect the obtained results. The venous blood was obtained after >3 hours fasting at two time-points; time-point 1 – at admission to the hospital; time-point 2 – on day 5 from stroke onset. The blood for control purposes was collected only once, and the blood collection at time-point 1 was done before thrombolytic treatment (nine patients. The amino acids were identified using the Amino Acids Analyser (AAA 400 by INGOS Corp., Praha, Czech Republic. Our results revealed a statistically significant increase of glutamate, cystine and methionine on day 1 of stroke, in comparison to control, whereas, proline level was decreased on day 1 of stroke – in comparison to control serum. On comparing day 5 to the initial day of IS, elevation was observed of levels of asparagine, glycine, tyrosine, arginine, threonine, valine, leucine and phenylalanine. It can be said, then, that ischemic stroke induces both essential and nonessential amino acid fluctuations. Moreover, the decrease in proline and glutamine serum level with the simultaneous increase in the concentration of branch chain amino acids, Glu and Thr suggests a violent mobilization of the body’s proteins. Thus, a decrease of Pro and a simultaneous increase of Glu serum level could be considered as a marker of acute IS.

Bacillus anthracis Overcomes an Amino Acid Auxotrophy by Cleaving Host Serum Proteins

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Terwilliger, Austen; Swick, Michelle C.; Pflughoeft, Kathryn J.; Pomerantsev, Andrei; Lyons, C. Rick; Koehler, Theresa M.

2015-01-01

ABSTRACT Bacteria sustain an infection by acquiring nutrients from the host to support replication. The host sequesters these nutrients as a growth-restricting strategy, a concept termed “nutritional immunity.” Historically, the study of nutritional immunity has centered on iron uptake because many bacteria target hemoglobin, an abundant circulating protein, as an iron source. Left unresolved are the mechanisms that bacteria use to attain other nutrients from host sources, including amino acids. We employed a novel medium designed to mimic the chemical composition of human serum, and we show here that Bacillus anthracis, the causative agent of anthrax disease, proteolyzes human hemoglobin to liberate essential amino acids which enhance its growth. This property can be traced to the actions of InhA1, a secreted metalloprotease, and extends to at least three other serum proteins, including serum albumin. The results suggest that we must also consider proteolysis of key host proteins to be a way for bacterial pathogens to attain essential nutrients, and we provide an experimental framework to determine the host and bacterial factors involved in this process. IMPORTANCE The mechanisms by which bacterial pathogens acquire nutrients during infection are poorly understood. Here we used a novel defined medium that approximates the chemical composition of human blood serum, blood serum mimic (BSM), to better model the nutritional environment that pathogens encounter during bacteremia. Removing essential amino acids from BSM revealed that two of the most abundant proteins in blood—hemoglobin and serum albumin—can satiate the amino acid requirement for Bacillus anthracis, the causative agent of anthrax. We further demonstrate that hemoglobin is proteolyzed by the secreted protease InhA1. These studies highlight that common blood proteins can be a nutrient source for bacteria. They also challenge the historical view that hemoglobin is solely an iron source for

Low normal thyroid function attenuates serum alanine aminotransferase elevations in the context of metabolic syndrome and insulin resistance in white people

NARCIS (Netherlands)

Dullaart, Robin P. F.; van den Berg, Eline H.; van der Klauw, Melanie; Blokzijl, Hans

Objectives: Thyroid hormones play a key role in hepatic lipid metabolism. Although hypothyroidismis associated with increased prevalence of non-alcoholic fatty liver disease (NAFLD), the relationship of NAFLD with low normal thyroid function is unclear. We tested the association of serum alanine

Pancreatic stellate cells support tumour metabolism through autophagic alanine secretion.

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Sousa, Cristovão M; Biancur, Douglas E; Wang, Xiaoxu; Halbrook, Christopher J; Sherman, Mara H; Zhang, Li; Kremer, Daniel; Hwang, Rosa F; Witkiewicz, Agnes K; Ying, Haoqiang; Asara, John M; Evans, Ronald M; Cantley, Lewis C; Lyssiotis, Costas A; Kimmelman, Alec C

2016-08-25

Pancreatic ductal adenocarcinoma (PDAC) is an aggressive disease characterized by an intense fibrotic stromal response and deregulated metabolism. The role of the stroma in PDAC biology is complex and it has been shown to play critical roles that differ depending on the biological context. The stromal reaction also impairs the vasculature, leading to a highly hypoxic, nutrient-poor environment. As such, these tumours must alter how they capture and use nutrients to support their metabolic needs. Here we show that stroma-associated pancreatic stellate cells (PSCs) are critical for PDAC metabolism through the secretion of non-essential amino acids (NEAA). Specifically, we uncover a previously undescribed role for alanine, which outcompetes glucose and glutamine-derived carbon in PDAC to fuel the tricarboxylic acid (TCA) cycle, and thus NEAA and lipid biosynthesis. This shift in fuel source decreases the tumour’s dependence on glucose and serum-derived nutrients, which are limited in the pancreatic tumour microenvironment. Moreover, we demonstrate that alanine secretion by PSCs is dependent on PSC autophagy, a process that is stimulated by cancer cells. Thus, our results demonstrate a novel metabolic interaction between PSCs and cancer cells, in which PSC-derived alanine acts as an alternative carbon source. This finding highlights a previously unappreciated metabolic network within pancreatic tumours in which diverse fuel sources are used to promote growth in an austere tumour microenvironment.

Determination of Selected Amino Acids in Serum of Patients with Liver Disease.

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Kanďár, Roman; Drábková, Petra; Toiflová, Tereza; Čegan, Alexander

2016-01-01

The determination of amino acids can be a reliable approach for extended diagnosis of liver diseases. This is because liver disease can be a cause of impaired amino acid metabolism. Therefore, a method for the determination of serum amino acids, applicable for clinical purposes, is necessary. The aim of this study was to find differences in the levels of selected amino acids between patients with liver disease and a control group. Samples of peripheral venous blood were obtained from a group of patients with liver disease (n = 131, 59 women at an average age of 60 years and 72 men at an average age of 52 years) and a control group (n = 105, 47 women at an average age of 62 years and 58 men at an average age of 58 years). Before the separation, the amino acids were derivatized with naphthalene-2,3-dicarboxaldehyde. For the separation, reverse phase column was used. The effluent was monitored with a fluorescence detector. There were significant differences in the concentrations of some amino acids between the patients and the control group, but also between women and men. Correlations between some amino acids and markers of liver blood tests and lipid metabolism were observed. A simple, relatively rapid and selective HPLC method with fluorescence detection for the determination of selected amino acids in serum has been developed.

Role of stimulated amino acid transport in promoting glycogenesis in the irradiated rat

International Nuclear Information System (INIS)

Kilberg, M.S.; Neuhaus, O.W.

1976-01-01

Whole-body irradiation of rats stimulates an amino acid transport system in the liver. Another phenomenon observed after exposure to ionizing radiations is the accumulation of hepatic glycogen. The data presented here relate the increased hepatic uptake of amino acids to glycogenesis. Male rats were exposed to two doses of γ rays, 2500 and 1500 R. Following exposure to 2500 R, the hepatic free amino acids were elevated during the first 48 hr accompanied by a decline in serum levels. At 72 hr the hepatic amino acids diminished to the control levels while the serum increased abruptly. By contrast, 72 hr after exposure to 1500 R the serum amino acid levels increased only 27 percent and the hepatic amino acid values increased 52 percent. These results are explained on the basis of the changes in AIB transport previously reported. The incorporation of 14 C from labeled L-alanine into hepatic glycogen was maximal 48 hr postexposure to 2500 R but declined to below control values at 72 hr. On the other hand, exposure to 1500 R resulted in maximal incorporation of 14 C at both 48 and 72 hr. We propose that transport of amino acids into liver cells is stimulated by the elevated blood levels of amino acids released from the degradation of protein. The transport increases the levels of hepatic free amino acids, and therefore, is a key factor in regulating postirradiation glycogenesis

Involvement of tyrosine residues, N-terminal amino acids, and beta-alanine in insect cuticular sclerotization.

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Andersen, Svend Olav

2007-09-01

During sclerotization of insect cuticle the acyldopamines, N-acetyldopamine (NADA) and N-beta-alanyldopamine (NBAD), are oxidatively incorporated into the cuticular matrix, thereby hardening and stabilizing the material by forming crosslinks between the proteins in the cuticular matrix and by forming polymers filling the intermolecular spaces in the cuticle. Sclerotized cuticle from the locust, Schistocerca gregaria, and the beetle, Tenebrio molitor, was hydrolyzed in dilute hydrochloric acid, and from the hydrolysates some components presumably degradation products of cuticular crosslinks were isolated. In two of the components, the sidechain of 3,4-dihydroxyacetophenone was linked to the amino groups of glycine and beta-alanine, respectively, and in the third component to the phenolic group of tyrosine. These three compounds, glycino-dihydroxyacetophenone, beta-alanino-dihydroxyacetophenone, and O-tyrosino-dihydroxyacetophenone, as well as the previously reported compound, lysino-dihydroxyacetophenone [Andersen, S.O., Roepstorff, P., 2007. Aspects of cuticular sclerotization in the locust, Schistocerca gregaria, and the beetle, Tenebrio molitor. Insect Biochem. Mol. Biol. 37, 223-234], are suggested to be degradation products of cuticular crosslinks, in which amino acid residues formed linkages to both the alpha- and beta-positions of the sidechain of acyldopamines.

Investigation of Mixed Chiral Selectors of Different Metal Ion-L-Alanine Complex and β-Cyclodextrin on the Chiral Separation of Dansyl Amino Acids with Capillary Electrophoresis

Institute of Scientific and Technical Information of China (English)

郑志侠; 屈锋; 林金明

2003-01-01

Chiral separation of dausyl amino acids by capillary electrophoresis using mixed selectors of Mn(ll)-L-alanine complex and β-cyclodextrin (β-CD) was studied. Resolution was considerably superior to that obtained by using either Mn (Ⅱ)-L-alanine complex or β-CD alone. The effects of separation parameters, such as pH value of buffer solution, capillary temperature, the concentration of Mn (Ⅱ)-L-alanine complex, the types of CD and ligand on the migration times and resolutions were investigated. Six different transition metal complexes,Cu(Ⅱ), Zn(Ⅱ), Co(Ⅱ), Ni(Ⅱ), Hg(Ⅱ) and Cd(Ⅱ)-L-alanine complexes have been employed and compared with Mn(Ⅱ)complex. Differences in retention and selectivity were found.The substitution of Cu(Ⅱ), Zn(Ⅱ), Co(Ⅱ) and Ni(Ⅱ) for Mn(Ⅱ) resulted in a better chiral resolution while Hg(Ⅱ) and Cd(Ⅱ) showed poorer resolution abilities. The chiral separation mechanism was also discussed briefly.

L-alanine uptake in membrane vesicles from Mytilus edulis gills

International Nuclear Information System (INIS)

Pajor, A.M.; Wright, S.H.

1986-01-01

Previous studies have shown that gills from M. edulis can accumulate L-alanine from seawater by a saturable process specific for α-neutral amino acids. This uptake occurs against chemical gradients in excess of 10 6 to 1. To further characterize this uptake, membrane vesicles were prepared from M. edulis gill tissue by differential centrifugation. Enrichments of putative enzyme markers (relative to that in combined initial fractions) were as follows: γ-Glutamyltranspeptidase, 25-30x; Alkaline Phosphatase, 5-6x; K + -dependent para-Nitrophenyl Phosphatase, 3-5x; Succinate Dehydrogenase 0.1-0.2x. These results suggest that the preparation is enriched in plasma membranes, although histochemical studies will be needed to verify this. The time course of 14 C-L-alanine uptake in the presence of inwardly-directed Na + gradient showed a transient overshoot (3-5 fold) at 10 minutes which decreased to equilibrium after six hours. The size of the overshoot and early uptake rates depended on the size of the inwardly-directed Na + gradient. No overshoot was seen in the presence of inwardly-directed gradients of LiCl or choline-Cl, or with equilibrium concentrations NaCl or mannitol. A reduced overshoot was seen with a gradient of NaSCN. A small overshoot was seen with an inwardly-directed gradient of KCl. Transport of L-alanine included saturable and diffusive components. Uptake of 6 μM L-alanine was inhibited more than 80% by 100 μM α-zwitterionic amino acids (alanine, leucine, glycine); by 30 to 75% by proline, aspartate and lysine; and less than 20% by a β-amino acid, taurine. The results of these experiments agree with those from intact gill studies and support the hypothesis that L-alanine is transported into gill epithelial cells by a secondary active transport process involving Na +

High serum carotenoids are associated with lower risk for developing elevated serum alanine aminotransferase among Japanese subjects: the Mikkabi cohort study.

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Sugiura, Minoru; Nakamura, Mieko; Ogawa, Kazunori; Ikoma, Yoshinori; Yano, Masamichi

2016-04-01

Many recent studies have shown that antioxidant vitamins and/or carotenoids may reduce liver disease, but this association has not been well established with thorough longitudinal cohort studies. The objective of this study was to longitudinally investigate whether serum carotenoids at baseline are associated with the risk of developing elevated serum alanine aminotransferase (ALT) among Japanese subjects. We conducted a follow-up study of 1073 males and females aged between 30 and 79 years at baseline from the Mikkabi prospective cohort study. Those who participated in the baseline study and completed follow-up surveys were examined longitudinally. Exclusions included excessive alcohol consumption (≥60 g alcohol/d), hepatitis B and C and having a history of medication use for liver disease. A cohort of 213 males and 574 females free of elevated serum ALT (>30 IU/ml) at baseline was studied. Over a mean follow-up period of 7·4 (sd 3·1) years, thirty-one males and forty-nine females developed new elevated serum ALT. After adjustments for confounders, the hazard ratios for elevated serum ALT in the highest tertiles of basal serum β-carotene, β-cryptoxanthin and total provitamin A carotenoids against the lowest tertiles were 0·43 (95 % CI 0·22, 0·81), 0·51 (CI 0·27, 0·94) and 0·52 (CI 0·28, 0·97), respectively. For α-carotene and lycopene, borderline reduced risks were also observed; however, these were not significant. Our results further support the hypothesis that antioxidant carotenoids, especially provitamin A carotenoids, might help prevent earlier pathogenesis of non-alcoholic liver disease in Japanese subjects.

Changes in Serum Free Amino Acids and Muscle Fatigue Experienced during a Half-Ironman Triathlon.

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Francisco Areces

Full Text Available The aim of this study was to investigate the relationship between changes in serum free amino acids, muscle fatigue and exercise-induced muscle damage during a half-ironman triathlon. Twenty-six experienced triathletes (age = 37.0 ± 6.8 yr; experience = 7.4 ± 3.0 yr competed in a real half-ironman triathlon in which sector times and total race time were measured by means of chip timing. Before and after the race, a countermovement jump and a maximal isometric force test were performed, and blood samples were withdrawn to measure serum free amino acids concentrations, and serum creatine kinase levels as a blood marker of muscle damage. Total race time was 320 ± 37 min and jump height (-16.3 ± 15.2%, P 20%. However, neither the changes in serum free amino acids nor the tryptophan/BCAA ratio were related muscle fatigue or muscle damage during the race.

The application of Alanine/ESR dosimetry in nuclear power plants

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Choi, Hoon; Lee, Byung Il [Radiation Health Research Institute, Seoul (Korea, Republic of)

2014-05-15

Alanine/Electron spin resonance(ESR) has been proven very effective tool which dosimetric characteristics is better suitable than generally used personnel dosimeter for long term dose estimation. L-α-alanine has unusual stability of radiation induced radicals. The fading is known as about 1% a year by IAEA technical documentary. It also has linear signal response on gamma rays over the wide range of dose quantity. Alanine is a kind of unnecessary amino acid in 20 amino acids. Estimation of the accumulated gamma dose is important to predict the life expectancy of cables. However, exact estimation of gamma dose at containment building of NPP is very difficult, because the variability of estimation value is apparently depending on the each installation position in containment building. Especially, some installation positions near reactor change extremely. So, the data from ESR measurement should be checked to the details on referring installation map and pictures.

Lack of Effect of Sodium Benzoate at Reported Clinical Therapeutic Concentration on d-Alanine Metabolism in Dogs.

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Popiolek, Michael; Tierney, Brendan; Steyn, Stefanus J; DeVivo, Michael

2018-06-19

Cognitive decline and psychosis have been hypothesized to be mediated by N-methyl-d-aspartate receptor (NMDAR) hypofunction. Consistent with this hypothesis, chronic treatment with d-alanine, a coagonist at the glycine site of the NMDAR, leads to an improvement of positive and cognitive symptoms in schizophrenic patients. d-alanine is oxidized by d-amino acid oxidase (DAAO); thus, an inhibitor of DAAO would be expected to enhance d-alanine levels and likewise lead to desirable clinical outcomes. Sodium benzoate, on the basis of d-amino acid inhibition, was observed to display beneficial clinical effects in schizophrenic and Alzheimer's patients. However, in the clinical pilot studies using sodium benzoate, d-amino acids were not quantified to verify that sodium benzoate's efficacy was mediated through DAAO inhibition. In this study, d-alanine content was monitored in cerebral spinal fluid (CSF) of dogs treated with daily injections of d-alanine (30 mg/kg) alone and in combination with sodium benzoate (30 mg/kg) for seven consecutive days. We reasoned that the cerebral spinal fluid d-alanine quantity is reflective of the brain d-alanine levels and it would increase as a consequence of DAAO inhibition with sodium benzoate. We found that d-alanine treatment lead to maximal concentration of 7.51 μM CSF d-alanine level; however, coadministration of sodium benzoate and d-alanine did not change CSF d-alanine level beyond that of d-alanine treatment alone. As a consequence, we conclude that clinical efficacy associated with chronic administration of sodium benzoate in schizophrenic and Alzheimer's patients is likely not mediated through inhibition of DAAO.

Reproducibility and signal response linearity of Alanine gel dosimeter

International Nuclear Information System (INIS)

Silva, Cleber Feijo Silva; Campos, Leticia Lucente

2008-01-01

Gel Dosimetry has been studied mainly for medical applications, because it presents signal response in the dose range used in radiotherapy treatments and it can be applied for three dimensional dosimetry. Alanine gel dosimeter is a new gel material developed at IPEN that presents significant improvement on previous alanine systems developed by Costa (1994). The DL-Alanine (C 3 H 7 NO 2 ) is an amino acid tissue equivalent that improves the production of ferric ions in the solution. These ferric ions concentration can be measured by spectrophotometry technique. This work aims to study the reproducibility of the alanine gel solutions and the signal response as a function of gamma radiation dose, considering that these two properties are very important for characterizing and standardizing any dosimeter. (author)

Hydrogen bonds in crystalline d-alanine: diffraction and spectroscopic evidence for differences between enantiomers

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Ezequiel A. Belo

2018-01-01

Full Text Available Enantiomeric amino acids have specific physiological functions in complex biological systems. Systematic studies focusing on the solid-state properties of d-amino acids are, however, still limited. To shed light on this field, structural and spectroscopic studies of d-alanine using neutron powder diffraction, polarized Raman scattering and ab initio calculations of harmonic vibrational frequencies were carried out. Clear changes in the number of vibrational modes are observed as a function of temperature, which can be directly connected to variations of the N—D bond lengths. These results reveal dissimilarities in the structural properties of d-alanine compared with l-alanine.

Complexation of vanadium (v) with alanine in different ionic strength

International Nuclear Information System (INIS)

Garib, F.; Zare, K.; Fekri, H

2002-01-01

The formation constants of species formed in the system H ++ alanine and VO 2 + alanine have be determined in aqueous solution for 1.0 3 NaCIO 4 ,using a combination of pramiracetam and spectrophotometric techniques. The compositions of the formed complexes and their stability constants were determined ny curve fitting method and it was shown that di oxovanadium(V) forms two mononuclear 1:1 and 1:2 species with alanine of the type VO 2 L and VO 2 L 2 The protonation constant of the amino group of alanine has been determined using a computer program which employ a least-squares method. The defence of the protonation of alanine and the stability constant of the species on ionic strength are described by a Debby-huckel type equation

Physiological hypercortisolemia increases proteolysis, glutamine, and alanine production

International Nuclear Information System (INIS)

Darmaun, D.; Matthews, D.E.; Bier, D.M.

1988-01-01

Physiological elevations of plasma cortisol levels, as are encountered in stress and severe trauma, were produced in six normal subjects by infusing them with hydrocortisone for 64 h. Amino acid kinetics were measured in the postabsorptive state using three 4-h infusions of L-[1- 13 C]leucine, L-phenyl[ 2 H 5 ]phenylalanine, L-[2- 15 N]glutamine, and L-[1- 13 C]alanine tracers (1) before, (2) at 12 h, and (3) at 60 h of cortisol infusion. Before and throughout the study, the subjects ate a normal diet of adequate protein and energy intake. The cortisol infusion raised plasma cortisol levels significantly from 10 ± 1 to 32 ± 4 μg/dl, leucine flux from 83 ± 3 to 97 ± 3 μmol·kg -1 ·h -1 , and phenylalanine flux from 34 ± 1 to 39 ± 1 (SE) μmol·kg -1 ·h -1 after 12 h of cortisol infusion. These increases were maintained until the cortisol infusion was terminated. These nearly identical 15% increases in two different essential amino acid appearance rates are reflective of increased whole body protein breakdown. Glutamine flux rose by 12 h of cortisol infusion and remained elevated at the same level at 64 h. The increase in flux was primarily due to a 55% increase in glutamine de novo synthesis. Alanine flux increased with acute hypercortisolemia and increased further at 60 h of cortisol infusion, a result primarily of increased alanine de novo synthesis. Insulin, alanine, and lactate plasma levels responded similarly with significant rises between the acute and chronic periods of cortisol infusion. Thus hypercortisolemia increases both protein breakdown and the turnover of important nonessential amino acids for periods of up to 64 h

Effect of streptococcal preparation (picibanil) on the postoperative rise in serum alanine aminotransferase activity in patients with urogenital cancer.

OpenAIRE

Taketa, Kazuhisa; Ohmori, Hiroyuki; Matsumura, Yonesuke; Asahi, Toshihiko; Okimune, Masaaki

1980-01-01

The effect of Picibanil, a streptococcal agent, on the development of liver injury after operations for urogenital cancer was studied retrospectively in the light of serum alanine aminotransferase (ALT) activity. The series comprised 32 cases receiving Picibanil and 33 controls with otherwise comparable clinical backgrounds. Picibanil reduced the incidence of postoperative ALT rise over 50 U/l within 6 weeks but increased it thereafter. The increase in ALT activity after 6 weeks was relativel...

Conductivity of alanine solution for high level dosimetry

International Nuclear Information System (INIS)

Wieser, A.; Figel, M.; Regulla, D.F.

1993-01-01

The amino acid alanine is well known as a dosimetric detector material for high level dosimetry. Its application is based on the formation of radicals by ionising radiation. The free radicals are earlier detected by electron spin resonance (ESR) spectroscopy or chemically after dissolving the irradiated samples. Of all these methods the ESR/alanine system is the most advanced and is suggested for reference dosimetry. At present, however, the high cost of the system is a serious handicap for a large scale routine application in radiation plants. In this study the variation of electrical conductivity of L-alanine solution with applied dose is investigated in the range from 0.5-200 kGy. The conductivity was measured with a 50 MHz RF oscillator. This readout method is uncomplicated and may be suitable for routine application. The experiments were performed with L-alanine solution in glass ampoules. (Author)

Metabolite Profiling of Feces and Serum in Hemodialysis Patients and the Effect of Medicinal Charcoal Tablets.

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Liu, Sixiu; Liang, Shanshan; Liu, Hua; Chen, Lei; Sun, Lingshuang; Wei, Meng; Jiang, Hongli; Wang, Jing

2018-05-22

Recently, the colon has been recognized as an important source of various uremic toxins in patients with end stage renal disease. Medicinal charcoal tablets are an oral adsorbent that are widely used in patients with chronic kidney disease in China to remove creatinine and urea from the colon. A parallel fecal and serum metabolomics study was performed to determine comprehensive metabolic profiles of patients receiving hemodialysis (HD). The effects of medicinal charcoal tablets on the fecal and serum metabolomes of HD patients were also investigated. Ultra-performance liquid chromatography/mass spectrometry was used to investigate the fecal and serum metabolic profiles of 20 healthy controls and 31 HD patients before and after taking medicinal charcoal tablets for 3 months. There were distinct metabolic variations between the HD patients and healthy controls both in the feces and serum according to multivariate data analysis. Metabolic disturbances of alanine, aspartate and glutamate metabolism, arginine and proline metabolism figured prominently in the serum. However, in the feces, alterations of tryptophan metabolism, lysine degradation and beta-alanine metabolism were pronounced, and the levels of several amino acids (leucine, phenylalanine, lysine, histidine, methionine, tyrosine, and tryptophan) were increased dramatically. Nineteen fecal metabolites and 21 serum metabolites were also identified as biomarkers that contributed to the metabolic differences. Additionally, medicinal charcoal treatment generally enabled the serum and fecal metabolomes of the HD patients to draw close to those of the control subjects, especially the serum metabolic profile. Parallel fecal and serum metabolomics uncovered the systematic metabolic variations of HD patients, especially disturbances in amino acid metabolism in the colon. Medicinal charcoal tablets had an impact on the serum and fecal metabolomes of HD patients, but their exact effects still need to be studied further

Essential amino acids in the gluten-free diet and serum in relation to depression in patients with celiac disease.

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Nathalie J M van Hees

Full Text Available Celiac disease (CD is associated with an increased risk of major depressive disorder, possibly due to deficiencies in micronutrients in the gluten-free diet. We aimed to investigate whether essential amino acids (i.e., the precursors of serotonin, dopamine and other neurotransmitters are depleted in the diet and serum of CD patients with major depressive disorder.In a cross-sectional study we assessed dietary intake of amino acids and serum levels of amino acids, in 77 CD patients on a gluten-free diet and in 33 healthy controls. Major depressive disorder was assessed with structured interviews (using the Mini International Neuropsychiatric Interview Plus. Dietary intake was assessed using a 203-item food frequency questionnaire.Participants had a mean age of 55 years and 74% were women. The intake of vegetable protein was significantly lower in CD patients than in healthy controls (mean difference of 7.8 g/d; 95% CI: 4.7-10.8, as were serum concentrations of tyrosine, phenylalanine and tryptophan (all p < 0.005. However, within the CD patient group, the presence of major depressive disorder (n = 42 was not associated with intake or serum levels of essential amino acids.Patients with CD on a long-term gluten-free diet, with good adherence, consume significantly less vegetable protein than controls, and their serum levels of several essential amino acids were also lower. Despite its potential adverse effect, intake and serum levels of essential amino acids were not related to major depression.

A molecular receptor selective for zwitterionic alanine.

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Rubio, Omayra H; Taouil, Rachid; Muñiz, Francisco M; Monleón, Laura M; Simón, Luis; Sanz, Francisca; Morán, Joaquín R

2017-01-04

A molecular receptor has been synthesized joining an aza-crown ether with a chiral chromane which mimics the oxyanion hole of the enzymes. With this receptor an apolar host-guest complex with zwitterionic alanine has been achieved through the formation of up to seven H-bonds. This complex allows the extraction of aqueous alanine to a chloroform phase, while other natural amino acids are poorly extracted or are not extracted at all. Due to the chiral nature of the receptor, enantioselective extraction from the aqueous alanine solution to a chloroform phase takes place. X-Ray analysis combined with anisotropic effects, NOE and CD studies revealed the absolute configuration of both strong and weak complexes. Modelling studies also support the proposed structures. The presence of an oxyanion-hole motif in this structure was corroborated by X-ray diffraction studies.

High Energy Electron Dosimetry by Alanine/ESR Spectroscopy

International Nuclear Information System (INIS)

Chu, Sung Sil

1989-01-01

Dosimetry based on electron spin resonance(ESR) analysis of radiation induced free radicals in amino acids is relevant to biological dosimetry applications. Alanine detectors are without walls and are tissue equivalent. Therefore, alanine ESR dosimetry looks promising for use in the therapy level. The dose range of the alanine/ESR dosimetry system can be extended down to l Gy. In a water phantom the absorbed dose of electrons generated by a medical linear accelerator of different initial energies (6-21 MeV) and therapeutic dose levels(1-60 Gy) was measured. Furthermore, depth dose measurements carried out with alanine dosimeters were compared with ionization chamber measurements. As the results, the measured absorbed doses for shallow depth of initial electron energies above 15 MeV were higher by 2-5% than those calculated by nominal energy CE factors. This seems to be caused by low energy scattered beams generated from the scattering foil and electron cones of beam projecting device in medical linear accelerator

Sulfur amino acids and alanine on pyrite (100) by X-ray photoemission spectroscopy: Surface or molecular role?

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Sanchez-Arenillas, M.; Galvez-Martinez, S.; Mateo-Marti, E., E-mail: mateome@cab.inta-csic.es

2017-08-31

Highlights: • Surface annealing pretreatment on pyrite surfaces can select molecular adsorption. • Enriched monosulfide species on pyrite (100) surface favors NH{sub 2} adsorption form. • Enriching disulfide species on pyrite (100) surface promotes NH{sub 3}{sup +} adsorption form. • Unique structure of each aminoacid provides a particular fingerprint in the process. • Spectroscopy evidence, pretreatment surface processes drives molecular adsorption. - Abstract: This paper describes the first successful adsorption of the cysteine, cystine, methionine and alanine amino acids on the pyrite (100) surface under ultra-high vacuum conditions with crucial chemical adsorption parameters driving the process. We have demonstrated by X-ray photoemission spectroscopy (XPS) that the surface pretreatment annealing process on pyrite surfaces is a critical parameter driving surface reactivity. The presence of enriched monosulfide species on the pyrite (100) surface favours the amino acid NH{sub 2} chemical form, whereas a longer annealing surface pretreatment of over 3 h repairs the sulfur vacancies in the pyrite, enriching disulfide species on the pyrite surface, which promotes NH{sub 3}{sup +} adsorption due to the sulfur vacancies in the pyrite being replaced by sulfur atom dimers (S{sub 2}{sup 2−}) on the surface. Furthermore, even if the surface chemistry (monosulfide or disulfide species enrichment) is the main factor promoting a partial conversion from NH{sub 2} to NH{sub 3}{sup +} species, the unique chemical structure of each amino acid provides a particular fingerprint in the process.

Amino Acid Composition of Grape Cultivars ( Vitis Spp.) in Japan

OpenAIRE

Shiraishi, Shin-ichi; 白石, 眞一

1991-01-01

The concentration of free amino acids and other chemical constituents in 75 grape cultivars at maturity were examined. The amino acid concentrations were found to be in decreasing order of arginine, proline, threonine and alanine in Vitis vinifera cultivars and alanine, arginine, threonine and proline in Vitis labmsca cultivars. The concentration of total amino acid was 5,083 ,umol in ‘Campbell Early’ and 3,391 pmol in ‘Yates’, both Vitis labrusca and table grapes. The amounts of total amino ...

A new synthesis of [3-11C]pyruvic acid using alanine racemase

International Nuclear Information System (INIS)

Ikemoto, M.; Okamoto, E.; Sasaki, M.; Haradahira, T.; Omura, H.; Furuya, Y.; Suzuki, K.; Watanabe, Y.

1998-01-01

The synthesis of [3- 11 C]pyruvic acid was attempted by two reaction systems (A: alanine racemase and D-amino acid oxidase, B: alanine racemase and L-alanine dehydrogenase) utilizing a new thermostable enzyme, alanine racemase. Conversion rates from D,L-[3- 11 C]alanine to [3- 11 C]pyruvic acid were almost 100% in both methods. Similar results were obtained with immobilized enzymes packed in a single column. Furthermore, the same column could be used repeatedly without a remarkable decrease of the [3- 11 C]pyruvic acid yield. Various matrices were tested for the immobilizing enzyme, and Aminopropyl-CPG was concluded to be the most suitable since the loss of the enzyme activity was the least in the studied matrices

Mineralization of alanine enantiomers in soil treated with heavy metals and nutrients

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Pavel Formánek

2011-01-01

Full Text Available This work deals with the determination of the effect of heavy metals and nutrients applied to the soil on alanine enatiomers mineralization with the main focus on evaluating the effect on L/D alanine respiration rate ratio. This study was initiated because previous research works revealed a change in L/D amino acid respiration under acid- or heavy metal-stress in soil. Generally, D-amino acids artificially supplied to soil are less utilized by microorganisms compared with their L-enantiomers. Stress of soil microorganisms cause decreased discrimination of D-amino acids utilization. Also, previous research showed that an application of fertilizers or combinations of fertilizers may affect the mineralization rate of L-amino acids differently, compared with their D-enantiomers. The results of this study show, that the effect of both heavy metals and nutrients on the L/D ratio was not clear, increasing or decreasing this ratio. Further research is necessary to broaden this study.

Perturbations in amino acids and metabolic pathways in osteoarthritis patients determined by targeted metabolomics analysis.

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Chen, Rui; Han, Su; Liu, Xuefeng; Wang, Kunpeng; Zhou, Yong; Yang, Chundong; Zhang, Xi

2018-05-15

Osteoarthritis (OA) is a degenerative synovial joint disease affecting people worldwide. However, the exact pathogenesis of OA remains unclear. Metabolomics analysis was performed to obtain insight into possible pathogenic mechanisms and diagnostic biomarkers of OA. Ultra-high performance liquid chromatography-triple quadrupole mass spectrometry (UPLC-TQ-MS), followed by multivariate statistical analysis, was used to determine the serum amino acid profiles of 32 OA patients and 35 healthy controls. Variable importance for project values and Student's t-test were used to determine the metabolic abnormalities in OA. Another 30 OA patients were used as independent samples to validate the alterations in amino acids. MetaboAnalyst was used to identify the key amino acid pathways and construct metabolic networks describing their relationships. A total of 25 amino acids and four biogenic amines were detected by UPLC-TQ-MS. Differences in amino acid profiles were found between the healthy controls and OA patients. Alanine, γ-aminobutyric acid and 4-hydroxy-l-proline were important biomarkers distinguishing OA patients from healthy controls. The metabolic pathways with the most significant effects were involved in metabolism of alanine, aspartate, glutamate, arginine and proline. The results of this study improve understanding of the amino acid metabolic abnormalities and pathogenic mechanisms of OA at the molecular level. The metabolic perturbations may be important for the diagnosis and prevention of OA. Copyright © 2018 Elsevier B.V. All rights reserved.

Complex association of serum alanine aminotransferase with the risk of future cardiovascular disease in type 2 diabetes.

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Afarideh, Mohsen; Aryan, Zahra; Ghajar, Alireza; Noshad, Sina; Nakhjavani, Manouchehr; Baber, Usman; Mechanick, Jeffrey I; Esteghamati, Alireza

2016-11-01

We aimed to determine the prospective association between baseline serum levels of alanine aminotransferase (ALT) and the incident cardiovascular disease (CVD) in people with type 2 diabetes. In an open cohort setting, people with type 2 diabetes were followed for their first ever CVD presentation from 1995 to 2015. Statistical methods included Cox regression analysis for reporting of hazard ratios (HRs), artificial neural network modelings, and risk reclassification analyses. We found a nearly constant CVD hazard with baseline serum ALT levels below the 30 IU/L mark, whereas baseline serum ALT levels ≥ 30 IU/L remained an independent predictor of lower CVD rates in patients with type 2 diabetes in the final multivariate Cox proportional hazards regression model (HR: 0.204, 95%CI [0.060-0.689], p for trend value = 0.006). Age, male gender and fasting plasma insulin levels independently predicted baseline serum ALT ≥ 30 IU/L among the population cohort. Augmentation of serum ALT into the weighted Framingham risk score resulted in a considerable net reclassification improvement (NRI) of coronary heart disease (CHD) risk prediction in the study population (NRI = 9.05% (8.01%-10.22%), p value < 0.05). Serum ALT could successfully reclassify about 9% of the population with type 2 diabetes across the CHD-affected and CHD-free categories. Overall, our findings demonstrate a complex and nonlinear relationship for the risk of future CVD by baseline serum ALT levels in patients with type 2 diabetes. Further studies are warranted to confirm whether this complex association could be translated into a clearly visible U or J-shaped figure. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

Changes in the free amino acid composition with maturity of the noble cultivar of Vitis rotundifolia Michx. grape.

Science.gov (United States)

Lamikanra, O; Kassa, A K

1999-12-01

The changes in amino acid composition that occur with maturity of the Noble cultivar of the Vitis rotundifolia Michx. (muscadine) grape were determined by HPLC. Eighteen amino acids were identified. Histidine was the most prominent amino acid followed by alanine. The concentrations of most of the major amino acids (alanine, glycine, histidine, valine, isoleucine, aspartic acid, and serine) were highest at verasion. Glutamine and threonine contents dropped sharply after fruit set, while those of arginine and proline increased gradually with maturity and ripening. Tyrosine content increased gradually with maturity and ripening following a slight drop after fruit set. In ripe grapes, seeds contained most of the amino acids in mature grapes (50%) followed by the pulp (23%), the juice (15%), and the skin (11%). Alanine, histidine, and arginine were the principal amino acids identified in the juice. Alanine, histidine, arginine, valine, glutamine, aspartic acid, proline, serine, and threonine accounted for about 90% of the amino acids in the pulp. In seeds, alanine, proline, asparagine, and histidine accounted for over 55% of the amino acids, while alanine and histidine were found to be the predominant free amino acids in the skin. The profile indicates some differences in the changes in amino acid composition with berry maturity and relative amounts of amino acids present in muscadine compared to those in nonmuscadine grape species.

Rapid and precise measurement of serum branched-chain and aromatic amino acids by isotope dilution liquid chromatography tandem mass spectrometry.

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Ruiyue Yang

Full Text Available BACKGROUND: Serum branched-chain and aromatic amino acids (BCAAs and AAAs have emerged as predictors for the future development of diabetes and may aid in diabetes risk assessment. However, the current methods for the analysis of such amino acids in biological samples are time consuming. METHODS: An isotope dilution liquid chromatography tandem mass spectrometry (ID-LC/MS/MS method for serum BCAAs and AAAs was developed. The serum was mixed with isotope-labeled BCAA and AAA internal standards and the amino acids were extracted with acetonitrile, followed by analysis using LC/MS/MS. The LC separation was performed on a reversed-phase C18 column, and the MS/MS detection was performed via the positive electronic spray ionization in multiple reaction monitoring mode. RESULTS: Specific analysis of the amino acids was achieved within 2 min. Intra-run and total CVs for the amino acids were less than 2% and 4%, respectively, and the analytical recoveries ranged from 99.6 to 103.6%. CONCLUSION: A rapid and precise method for the measurement of serum BCAAs and AAAs was developed and may serve as a quick tool for screening serum BCAAs and AAAs in studies assessing diabetes risk.

Prebiotic Amino Acid Thioester Synthesis: Thiol-Dependent Amino Acid Synthesis from Formose substrates (Formaldehyde and Glycolaldehyde) and Ammonia

Science.gov (United States)

Weber, Arthur L.

1998-01-01

Formaldehyde and glycolaldehyde (substrates of the formose autocatalytic cycle) were shown to react with ammonia yielding alanine and homoserine under mild aqueous conditions in the presence of thiol catalysts. Since similar reactions carried out without ammonia yielded alpha-hydroxy acid thioesters, the thiol-dependent synthesis of alanine and homoserine is presumed to occur via amino acid thioesters-intermediates capable of forming peptides. A pH 5.2 solution of 20 mM formaldehyde, 20 mM glycolaldehyde, 20 mM ammonium chloride, 23 mM 3-mercaptopropionic acid, and 23 mM acetic acid that reacted for 35 days at 40 C yielded (based on initial formaldehyde) 1.8% alanine and 0.08% homoserine. In the absence of thiol catalyst, the synthesis of alanine and homoserine was negligible. Alanine synthesis required both formaldehyde and glycolaldehyde, but homoserine synthesis required only glycolaldehyde. At 25 days the efficiency of alanine synthesis calculated from the ratio of alanine synthesized to formaldehyde reacted was 2.1%, and the yield (based on initial formaldehyde) of triose and tetrose intermediates involved in alanine and homoserine synthesis was 0.3 and 2.1%, respectively. Alanine synthesis was also seen in similar reactions containing only 10 mM each of aldehyde substrates, ammonia, and thiol. The prebiotic significance of these reactions that use the formose reaction to generate sugar intermediates that are converted to reactive amino acid thioesters is discussed.

Changes in alanine turnover rate due to nutritional and genetic obesity in the rat.

Science.gov (United States)

Yebras, M; Salvadó, J; Arola, L; Remesar, X; Segués, T

1994-08-01

The changes in alanine turnover were determined in Zucker rats, which were either genetically obese (fa/fa) or rendered obese by dietary treatment (cafeteria fed). The whole body rate of alanine turnover was higher in genetically obese rats than in rats in which obesity was induced by diet (cafeteria). This is possibly due to variations in the rate of the amino acid incorporation into proteins, since the rate of whole body alanine degradation is the same for both groups. Thus, the different pattern followed by alanine turnover rate in these types of obese animals reflects the differences in the nitrogen economy of these animals, pointing to a higher alanine utilization in the genetically obese animals and a conservative management of alanine in the cafeteria-fed animals.

Effects of a honeybee sting on the serum free amino acid profile in humans.

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Jan Matysiak

Full Text Available The aim of this study was to assess the response to a honeybee venom by analyzing serum levels of 34 free amino acids. Another goal of this study was to apply complex analytic-bioinformatic-clinical strategy based on up-to-date achievements of mass spectrometry in metabolomic profiling. The amino acid profiles were determined using hybrid triple quadrupole/linear ion trap mass spectrometer coupled with a liquid chromatography instrument. Serum samples were collected from 27 beekeepers within 3 hours after they were stung and after a minimum of 6 weeks following the last sting. The differences in amino acid profiles were evaluated using MetaboAnalyst and ROCCET web portals. Chemometric tests showed statistically significant differences in the levels of L-glutamine (Gln, L-glutamic acid (Glu, L-methionine (Met and 3-methyl-L-histidine (3MHis between the two analyzed groups of serum samples. Gln and Glu appeared to be the most important metabolites for distinguishing the beekeepers tested shortly after a bee sting from those tested at least 6 weeks later. The role of some amino acids in the response of an organism to the honeybee sting was also discussed. This study indicated that proposed methodology may allow to identify the individuals just after the sting and those who were stung at least 6 weeks earlier. The results we obtained will contribute to better understanding of the human body response to the honeybee sting.

Neonatal taurine and alanine modulate anxiety-like behavior and decelerate cortical spreading depression in rats previously suckled under different litter sizes.

Science.gov (United States)

Francisco, Elian da Silva; Guedes, Rubem Carlos Araújo

2015-11-01

The amino acids taurine and alanine play a role in several physiological processes, including behavior and the electrical activity of the brain. In this study, we investigated the effect of treatment with taurine or alanine on anxiety-like behavior and the excitability-dependent phenomenon known as cortical spreading depression (CSD), using rats suckled in litters with 9 and 15 pups (groups L9 and L15). From postnatal days 7 to 27, the animals received per gavage 300 mg/kg/day of taurine or alanine or both. At 28 days, we tested the animals in the elevated plus maze, and at 33-35 days, we recorded CSD and analyzed its velocity of propagation, amplitude, and duration. Compared with water-treated controls, the L9 groups treated with taurine or alanine displayed anxiolytic behavior (higher number of entries in the open arms; p taurine, alanine, or both) treated at adulthood (90-110 days). The L15 condition resulted in smaller durations and higher CSD velocities compared with the L9 condition. Besides reinforcing previous evidence of behavioral modulation by taurine and alanine, our data are the first confirmation that treatment with these amino acids decelerates CSD regardless of lactation conditions (normal versus unfavorable lactation) or age at amino acid administration (young versus adult). The results suggest a modulating role for both amino acids on anxiety behavior and neuronal electrical activity.

The Origin of Amino Acids in Lunar Regolith Samples

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Cook, Jamie E.; Callahan, Michael P.; Dworkin, Jason P.; Glavin, Daniel P.; McLain, Hannah L.; Noble, Sarah K.; Gibson, Everett K., Jr.

2016-01-01

We analyzed the amino acid content of seven lunar regolith samples returned by the Apollo 16 and Apollo 17 missions and stored under NASA curation since collection using ultrahigh-performance liquid chromatography with fluorescence detection and time-of-flight mass spectrometry. Consistent with results from initial analyses shortly after collection in the 1970s, we observed amino acids at low concentrations in all of the curated samples, ranging from 0.2 parts-per-billion (ppb) to 42.7 ppb in hot-water extracts and 14.5 ppb to 651.1 ppb in 6M HCl acid-vapor-hydrolyzed, hot-water extracts. Amino acids identified in the Apollo soil extracts include glycine, D- and L-alanine, D- and L-aspartic acid, D- and L-glutamic acid, D- and L-serine, L-threonine, and L-valine, all of which had previously been detected in lunar samples, as well as several compounds not previously identified in lunar regoliths: -aminoisobutyric acid (AIB), D-and L-amino-n-butyric acid (-ABA), DL-amino-n-butyric acid, -amino-n-butyric acid, -alanine, and -amino-n-caproic acid. We observed an excess of the L enantiomer in most of the detected proteinogenic amino acids, but racemic alanine and racemic -ABA were present in some samples.

Effects of high grade bentonite on performance, organ weights and serum biochemistry during aflatoxicosis in broilers

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H. C. Indresh

2013-12-01

Full Text Available Aim: To evaluate the effect of different levels of High Bentonite on growth performance, organ weight and serum biochemistry in broiler fed on diets containing aflatoxin. Materials and Methods: A total of 360 day-old commercial broiler chicks were divided at random into 8 dietary treatment groups of 42 chicks each having 3 replicates. Dietary levels of aflatoxin (0.5 ppm and High-grade bentonite (0.5, 0.75 and 1.00% were tested in a completely randomized design manner, forming a total of 8 dietary treatments each with three replicates. Body weight and feed intake were recorded weekly. At 5 wk, six birds from each treatment were sacrificed and liver, kidney, gizzard, pancreas, spleen, bursa of Fabricius and thymus were extracted and weighed. The serum samples were analyzed for total proteins, uric acid, serum albumin, serum globulin and the activities of gamma glutamyl transferase (GGT, alanine amino transferase (ALT and for antibody titers against Newcastle disease (ND and infectious bursal disease (IBD using ELISA technique. Results: A significant (P<0.05 decrease in body weight, feed consumption, relative weights of bursa, thymus, serum protein, anti body titers against NDV and IBDV, and increase in FCR, mortality, relative weight of liver, kidney, and the activity of Gamma Glutamyl Transferase (GGT was observed. However, the relative weights of gizzard, pancreas, spleen, serum albumin, uric acid and the activity of Alanine Amino Transferase (ALT were not influenced by inclusion of AF or HGB. Conclusion: The addition of HGB restored the harmful effects of AF on body weight, feed consumption, FCR, mortality, relative weight of liver, kidney, serum protein, IBDV and NDV. Supplementation of high grade bentonite at 1.0 per cent level was found to be beneficial in ameliorating the adverse effects of aflatoxin (AF in broiler chickens. [Vet World 2013; 6(6.000: 313-317

LIMITED ELEVATIONS IN ANTITUBERCULOSIS DRUG-INDUCED SERUM ALANINE AMINOTRANSFERASE (ALT) LEVELS IN A COHORT OF NIGERIANS ON TREATMENT FOR PULMONARY TUBERCULOSIS AND HIV INFECTION IN YENAGOA

NARCIS (Netherlands)

Ikuabe, Peter Ogie; Ebuenyi, Ikenna Desmond; Harry, Tubonye Clement

2015-01-01

BACKGROUND: This study, undertaken in a major tertiary hospital in the Niger Delta region of Nigeria was designed to examine the incidence of elevation in serum alanine aminotransference (ALT) in our patients who were on treatment for HIV/AIDS with some of them on antituberculosis drugs. METHOD:

Dynamics of human whole body amino acid metabolism

International Nuclear Information System (INIS)

Young, V.R.

1981-01-01

The mechanism of regulation of the nitrogen metabolism in humans under various nutritional and physiological states was examined using stable isotopes. In the simultaneous continuous infusion of 1- [ 13 ] - leucine and α- [ 15 N]- lysine, their fluxed decreased when individuals received lower protein intake. The rates of oxidation and incorporation into body proteins of leucine changed in parallel with the protein intake. Such effects of diet on whole body leucine kinetics were modified by the energy state and dietary energy level. The nitrogen balance was also improved by an excess level of dietary energy. When the intake of dietary protein was lowered below the maintenance level, the whole body flux and de novo synthesis of glycine were lowered, but alanine synthesis was clearly increased. The intravenous infusion of glucose at 4 mg/kg.min, which causes increase in excess blood sugar and plasma insulin, increased the alanine flux, but had no effect on the glycine flux. The rate of albumin synthesis, determined by giving 15 N-glycine orally every 3 hr, decreased with the lowered intake of dietary protein in young men, but not in elderly men. This explains why the serum albumin synthesis increases with the increase in the intake of dietary protein in young men, but not in elderly men. The rate of whole body protein synthesis in young men receiving the L-amino acid diets providing with the required intake of specific amino acid was much lower than that in the men receiving the diets providing with generous intake of specific amino acid. Thus the control mechanism to maintain the homeostasis of body nitrogen and amino acids is related in some unknown way to the nutritional requirement of the hosts. (Kaihara, S.)

Postirradiation effects in alanine dosimeter probes of two different suppliers

International Nuclear Information System (INIS)

Anton, Mathias

2008-01-01

The measurand relevant for the dosimetry for radiation therapy is the absorbed dose to water, D W . The Physikalisch-Technische Bundesanstalt (PTB) is establishing a secondary standard for D W for high-energy photon and electron radiation based on electron spin resonance (ESR) of the amino acid alanine. For practical applications, like, for example, intercomparison measurements using the ESR/alanine dosimetry system, the temporal evolution of the ESR signal of irradiated probes is an important issue. This postirradiation behaviour is investigated for alanine pellets of two different suppliers for different storage conditions. The influence of the storage conditions on the temporal evolution may be dependent on the type of probes used. The measurement and analysis method developed at the PTB is able to circumvent the apparent difficulties in the case of alanine/paraffin probes. Care has to be taken in case this method cannot be applied

Survivability and reactivity of glycine and alanine in early oceans: effects of meteorite impacts.

Science.gov (United States)

Umeda, Yuhei; Fukunaga, Nao; Sekine, Toshimori; Furukawa, Yoshihiro; Kakegawa, Takeshi; Kobayashi, Takamichi; Nakazawa, Hiromoto

2016-01-01

Prebiotic oceans might have contained abundant amino acids, and were subjected to meteorite impacts, especially during the late heavy bombardment. It is so far unknown how meteorite impacts affected amino acids in the early oceans. Impact experiments were performed under the conditions where glycine was synthesized from carbon, ammonia, and water, using aqueous solutions containing (13)C-labeled glycine and alanine. Selected amino acids and amines in samples were analyzed with liquid chromatography-mass spectrometry (LC/MS). In particular, the (13)C-labeled reaction products were analyzed to distinguish between run products and contaminants. The results revealed that both amino acids survived partially in the early ocean through meteorite impacts, that part of glycine changed into alanine, and that large amounts of methylamine and ethylamine were formed. Fast decarboxylation was confirmed to occur during such impact processes. Furthermore, the formation of n-butylamine, detected only in the samples recovered from the solutions with additional nitrogen and carbon sources of ammonia and benzene, suggests that chemical reactions to form new biomolecules can proceed through marine impacts. Methylamine and ethylamine from glycine and alanine increased considerably in the presence of hematite rather than olivine under similar impact conditions. These results also suggest that amino acids present in early oceans can contribute further to impact-induced reactions, implying that impact energy plays a potential role in the prebiotic formation of various biomolecules, although the reactions are complicated and depend upon the chemical environments as well.

Hb and dyslipidaemia as predicting markers of serum alanine aminotransferase elevation in Chinese adolescents.

Science.gov (United States)

Chao, Kuo-Ching; Chang, Chun-Chao; Owaga, Eddy; Bai, Chyi-Huey; Huang, Tzu-chieh; Pan, Wen-Harn; Chang, Jung-Su

2016-04-01

Fe is an essential element for erythropoiesis and Hb synthesis. High Hb levels affect the blood's viscosity and are associated with cardiovascular dysfunction. The aim of the present study was to examine relationships of Hb and cardiometabolic abnormalities with the risk of alanine aminotransferase (ALT) elevation in adolescents. A population-based, cross-sectional study. National Nutrition and Health Survey in Taiwan (2010-2011, adolescents). Healthy adolescents aged 13-18 years. In total, 1941 adolescents (963 boys and 978 girls) were entered in the study. The mean age was 15·3 (sd 0·1) years (boys, 15·3 (sd 0·1) years; girls, 15·2 (sd 0·1) years). ALT tertile cut-off points for boys were 11 and 16 U/l, and for girls were 9 and 12 U/l. Girls without dyslipidaemia and presenting in the highest quartile (Q1) of Hb (>13·6 g/dl) were 1·89 and 3·76 times more likely to have raised serum ALT (9 and >12 U/l, respectively) than the reference (lowest quartile of Hb (Q1), 12 U/l) than the reference (Q1 of Hb, 15·4 g/dl), who were 7·40 times more likely to have elevated serum ALT of >16 U/l than the reference (Q1 of Hb, Hb level is a predictor of elevated serum ALT in adolescent girls with dyslipidaemia. Our study also highlights the importance of further research to establish cut-off points for Hb and its utility in diagnosing and preventing the onset of dyslipidaemia in adolescents.

Amino acid composition of cottage cheese and whey with bifidobacteria

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N. S. Rodionova

2017-01-01

Full Text Available Breaking condition intestinal flora, the simplest way to create sensitive parts of the body microbiocenosis caused by various factors. Reasons for people microecology imbalances – antibiotics, preservatives, stress. The result is an imbalance – the number of gastrointestinal disorders, immune deficiency disorders metabolic processes in the body. Therapeutic effect of probiotic microorganisms is the result of exogenous and endometabolitov synthesis the character of the protein. Acid activity information probiotic microorganisms and the distribution of amino acids between the products and intermediates in biopotential evaluation process foods produced with probiotic microflora by fermentation. Test results from the amino acid composition of whey and quark are obtained by fermenting raw milk probiotics bifidobacterias. It was found that during the fermentation of the quark consortium, Bifidobacterium bifidum, Bifidobacterium longum, Bifidobacterium adolescentis, Bifidobacterium bifidum Y enriched curd 4 of leucine and glutamine. Rate of transfer amino acids in serum to 2–6% of essential amino acids is irrelevant 3–7%. The highest value observed transient threonine, isoleucyl, lysine, valine, alanine, glycine, proline, serine. The mean value of the prototype amino acid protein curd and whey protein biological value was 71.89 and 74.58. Preservation of active forms of probiotic microorganisms after heating the bunch to 53–55 °C, lg concentration of not less than 7 (in 1 g in cottage cheese and serum. The received data are actual for formation of an information data bank, necessary for the development of prescription-component solutions of eubiotic products.

Experiences with alanine dosimetry in afterloading brachytherapy

International Nuclear Information System (INIS)

Eberhardt, H.-J.; Gohs, U.

1996-01-01

At the present, the most commonly used dosimetry for radiotherapy applications are ionisation chambers and thermoluminescent dosimeters (TLD). However, there are some undesirable characteristics of these dosimetry systems, such as large detection volume (ionisation chamber) as well as fading of the radiation induced signal with time and destructive readout (TLG). The present study is an investigation into the use of the alanine/ESR dosimetry in fractionated afterloading brachytherapy during the whole radiotherapy course. There are some qualities which make alanine dosimetry attractive. These are the linear energy response, low fading under standard conditions, and the nondestructive readout. Thus the alanine dosimetry makes possible cumulative dose measurements during the radiotherapy course and an archival storage. By ionizing radiation (gamma, e, n, p, charged particles) free radicals (unpaired electrons) are produced in the amino acid alanine. The continuous wave electron spin resonance (ESR) spectroscopy is used to determine the number of free radicals, which is proportional to the absorbed dose and the alanine content of the dosimeter. The ESR measurements were made at room temperature using a Bruker EPR analyzer EMS-104. The dosimeters used in the test are alanine pellets (23.72 mg weight, 4.9 mm diameter, 1 mm height) as well as flexible alanine film dosimeters (thickness about 500 μm). The dosimeters consist of a blend of L-alpha-alanine and a binder. The alanine content of the pellets and the film dosimeters is about 88 % and 50 % by weight, respectively. The dosimeters for the calculation of the dose-effect-relationship were irradiated at the Physical-Technical Bundesanstalt in Braunschweig by a standard 60Co source. The maximum deviation from the calculated linear function is about 0.12 Gy in the dose range up to 80 Gy. The goal of medical applications was the superficial dose measurement in afterloading brachytherapy during the radiotherapy course in

Effect of Amino Acid Substitutions in the GerAA Protein on the Function of the Alanine-Responsive Germinant Receptor of Bacillus subtilis Spores▿

Science.gov (United States)

Mongkolthanaruk, Wiyada; Cooper, Gareth R.; Mawer, Julia S. P.; Allan, Raymond N.; Moir, Anne

2011-01-01

Spores of Bacillus subtilis require the GerAA, GerAB, and GerAC receptor proteins for l-alanine-induced germination. Mutations in gerAA, both random and site directed, result in phenotypes that identify amino acid residues important for receptor function in broad terms. They highlight the functional importance of two regions in the central, integral membrane domain of GerAA. A P324S substitution in the first residue of a conserved PFPP motif results in a 10-fold increase in a spore's sensitivity to alanine; a P326S change results in the release of phase-dark spores, in which the receptor may be in an “activated” or “quasigerminated” state. Substitutions in residues 398 to 400, in a short loop between the last two likely membrane-spanning helices of this central domain, all affect the germination response, with the G398S substitution causing a temperature-sensitive defect. In others, there are wider effects on the receptor: if alanine is substituted for conserved residue N146, H304, or E330, a severe defect in l-alanine germination results. This correlates with the absence of GerAC, suggesting that the assembly or stability of the entire receptor complex has been compromised by the defect in GerAA. In contrast, severely germination-defective mutants such as E129K, L373F, S400F, and M409N mutants retain GerAC at normal levels, suggesting more local and specific effects on the function of GerAA itself. Further interpretation will depend on progress in structural analysis of the receptor proteins. PMID:21378197

Amino acid metabolism in plant leaf, 1

International Nuclear Information System (INIS)

Ito, Osamu; Kumazawa, Kikuo

1977-01-01

14C-labelled sodium bicarbonate and 15N-labelled ammonium sulfate were simultaneously vacuum-infiltrated into detached sunflower leaves, and the incorporation of 14C and 15N into free amino acids was chased during 60-min period in the light and in the dark. In the light, the 14C specific activity of aspartic acid, alanine, serine and glycine rapidly increased for 5 min and thereafter decreased. On the other hand, that of glutamic acid continued to increase slowly during the entire 60-min period. In the dark, aspartic acid most actively incorporated 14C. The difference of changes in 14C specific activity between glutamic acid and other amino acids was also observed in the dark as in the light. These results suggest that the carbon skeleton of glutamic acid is synthesized from aspartic acid, alanine, serine and glycine. 15N content of glutamine was the highest of all amino acids investigated in the light, and it was followed by glutamic acid, alanine, aspartic acid, serine and glycine, in this order. In the dark, 15N content of glutamic acid fell remarkably and was lower than that of alanine up to 5 min. From these 15N tracer experiments, it is suggested that the incorporation of ammonium into glutamic acid is strictly dependent on light and that alanine incorporates ammonium by the direct animation besides the transamination from glutamic acid. (auth.)

Elevated Serum Levels of Cysteine and Tyrosine: Early Biomarkers in Asymptomatic Adults at Increased Risk of Developing Metabolic Syndrome

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Nina Mohorko

2015-01-01

Full Text Available As there is effective intervention for delaying or preventing metabolic diseases, which are often present for years before becoming clinically apparent, novel biomarkers that would mark metabolic complications before the onset of metabolic disease should be identified. We investigated the role of fasting serum amino acids and their associations with inflammatory markers, adipokines, and metabolic syndrome (MetS components in subjects prior to the onset of insulin resistance (IR. Anthropometric measurements, food records, adipokines, biochemical markers, and serum levels of amino acids were determined in 96 asymptomatic subjects aged 25–49 years divided into three groups according to the number of MetS components present. Cysteine and tyrosine were significantly higher already in group with one component of MetS present compared to subjects without MetS components. Serum amino acid levels correlated with markers of inflammation and adipokines. Alanine and glycine explained 10% of insulin resistance variability. The role of tyrosine and cysteine, that were higher already with 1 component of MetS present, should be further investigated as they might point to future insulin disturbances.

Rapid and Precise Measurement of Serum Branched-Chain and Aromatic Amino Acids by Isotope Dilution Liquid Chromatography Tandem Mass Spectrometry

OpenAIRE

Yang, Ruiyue; Dong, Jun; Guo, Hanbang; Li, Hongxia; Wang, Shu; Zhao, Haijian; Zhou, Weiyan; Yu, Songlin; Wang, Mo; Chen, Wenxiang

2013-01-01

BACKGROUND: Serum branched-chain and aromatic amino acids (BCAAs and AAAs) have emerged as predictors for the future development of diabetes and may aid in diabetes risk assessment. However, the current methods for the analysis of such amino acids in biological samples are time consuming. METHODS: An isotope dilution liquid chromatography tandem mass spectrometry (ID-LC/MS/MS) method for serum BCAAs and AAAs was developed. The serum was mixed with isotope-labeled BCAA and AAA internal standar...

The Amino Acid Composition of the Sutter's Mill Carbonaceous Chondrite

Science.gov (United States)

Glavin, D. P.; Burton, A. S.; Elsila, J. E.; Dworkin, J. P.; Yin, Q. Z.; Cooper, G.; Jenniskens, P.

2012-01-01

In contrast to the Murchison meteorite which had a complex distribution of amino acids with a total C2 to Cs amino acid abundance of approx.14,000 parts-per-billion (ppb) [2], the Sutters Mill meteorite was found to be highly depleted in amino acids. Much lower abundances (approx.30 to 180 ppb) of glycine, beta-alanine, L-alanine and L-serine were detected in SM2 above procedural blank levels indicating that this meteorite sample experienced only minimal terrestrial amino acid contamination after its fall to Earth. Carbon isotope measurements will be necessary to establish the origin of glycine and beta-alanine in SM2. Other non-protein amino acids that are rare on Earth, yet commonly found in other CM meteorites such as aaminoisobutyric acid (alpha-AIB) and isovaline, were not identified in SM2. However, traces of beta-AIB (approx.1 ppb) were detected in SM2 and could be" extraterrestrial in origin. The low abundances of amino acids in the Sutter's Mill meteorite is consistent with mineralogical evidence that at least some parts of the Sutter's Mill meteorite parent body experienced extensive aqueous and/or thermal alteration.

Effect of the ionizing radiation on alanine solution for a dosimeter application

International Nuclear Information System (INIS)

Abdessamad, Nour El Houda

2007-01-01

The amino acid alanine is well known as a dosimetric detector material for high level dosimetry. Its application is based on the formation of radicals by ionising radiation. In this study the effect of several parameters such as: the ionising radiation, the concentration, the dose on the pH, conductivity and the oscillotitrometric answer of L a lanine solution was investigated. The results show that there is a significant production of new species. The formation of these species increases upon increasing dose. The comparison between the repeatability of the used techniques led us to choose of the system alanine/pH and the alanine/conductivity as the most adapted. (Author)

Alanine Radiation Detectors in Therapeutic Carbon Ion Beams

DEFF Research Database (Denmark)

Herrmann, Rochus; Jäkel, Oliver; Palmans, Hugo

of the depth dose curves. Solid state detectors, such as diamond detectors, radiochromic films, TLDs and the amino acid alanine are used due to there good spatial resolution. If used in particle beams their response often exhibits a dependence on particle energy and type, so the acquired signal is not always...... proportional to absorbed dose. A model by Hansen and Olsen, based on the Track Structure Theory is available, which can predict the relative efficiency of some detectors, when the particle spectrum is known. For alanine detectors the model was successfully validated by Hansen and Olsen for several ion species...... at energies below 20 MeV/u. We implemented this model in the Monte Carlo code FLUKA. At the GSI heavy ion facility in Darmstadt, Germany, alanine has been irradiated with carbon ions at energies between 88 an 400 MeV/u, which is the energy range used for therapy. The irradiation and the detector response have...

A Clostridium difficile alanine racemase affects spore germination and accommodates serine as a substrate.

Science.gov (United States)

Shrestha, Ritu; Lockless, Steve W; Sorg, Joseph A

2017-06-23

Clostridium difficile has become one of the most common bacterial pathogens in hospital-acquired infections in the United States. Although C. difficile is strictly anaerobic, it survives in aerobic environments and transmits between hosts via spores. C. difficile spore germination is triggered in response to certain bile acids and glycine. Although glycine is the most effective co-germinant, other amino acids can substitute with varying efficiencies. Of these, l-alanine is an effective co-germinant and is also a germinant for most bacterial spores. Many endospore-forming bacteria embed alanine racemases into their spore coats, and these enzymes are thought to convert the l-alanine germinant into d-alanine, a spore germination inhibitor. Although the C. difficile Alr2 racemase is the sixth most highly expressed gene during C. difficile spore formation, a previous study reported that Alr2 has little to no role in germination of C. difficile spores in rich medium. Here, we hypothesized that Alr2 could affect C. difficile l-alanine-induced spore germination in a defined medium. We found that alr2 mutant spores more readily germinate in response to l-alanine as a co-germinant. Surprisingly, d-alanine also functioned as a co-germinant. Moreover, we found that Alr2 could interconvert l- and d-serine and that Alr2 bound to l- and d-serine with ∼2-fold weaker affinity to that of l- and d-alanine. Finally, we demonstrate that l- and d-serine are also co-germinants for C. difficile spores. These results suggest that C. difficile spores can respond to a diverse set of amino acid co-germinants and reveal that Alr2 can accommodate serine as a substrate. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

Breed and species comparison of amino acid transport variation in equine erythrocytes.

Science.gov (United States)

Fincham, D A; Young, J D; Mason, D K; Collins, E A; Snow, D H

1985-05-01

The amino acid permeability of red blood cells from Equus caballus (thoroughbred, Arab, shire and pony), E przewalskii (Przewalski's horse), E asinus (donkey and mule) and E burchelli (common or plains zebra) was measured. Individual animals exhibited stable but widely differing rates of L-[U-14C]alanine uptake in the range 5 to 1554 mumol (litre cells)-1 h-1 (0.2 mM extracellular L-alanine, 37 degrees C). Of the thoroughbreds tested, 30 per cent had red blood cells which were essentially impermeable to L-alanine (5 to 10 mumol (litre cells)-1 h-1, giving transport rates similar to those found previously in amino acid transport-deficient sheep erythrocytes. In contrast, only 3 per cent of the ponies tested had red blood cells impermeable to L-alanine. No cases of erythrocyte amino acid transport deficiency were found in the other horse breeds and species tested.

Forced swimming and imipramine modify plasma and brain amino acid concentrations in mice.

Science.gov (United States)

Murakami, Tatsuro; Yamane, Haruka; Tomonaga, Shozo; Furuse, Mitsuhiro

2009-01-05

The relationships between monoamine metabolism and forced swimming or antidepressants have been well studied, however information is lacking regarding amino acid metabolism under these conditions. Therefore, the aim of the present study was to investigate the effects of forced swimming and imipramine on amino acid concentrations in plasma, the cerebral cortex and the hypothalamus in mice. Forced swimming caused cerebral cortex concentrations of L-glutamine, L-alanine, and taurine to be increased, while imipramine treatment caused decreased concentrations of L-glutamate, L-alanine, L-tyrosine, L-methionine, and L-ornithine. In the hypothalamus, forced swimming decreased the concentration of L-serine while imipramine treatment caused increased concentration of beta-alanine. Forced swimming caused increased plasma concentration of taurine, while concentrations of L-serine, L-asparagine, L-glutamine and beta-alanine were decreased. Imipramine treatment caused increased plasma concentration of all amino acid, except for L-aspartate and taurine. In conclusion, forced swimming and imipramine treatment modify central and peripheral amino acid metabolism. These results may aid in the identification of amino acids that have antidepressant-like effects, or may help to refine the dosages of antidepressant drugs.

Carbohydrate metabolism during prolonged exercise and recovery: interactions between pyruvate dehydrogenase, fatty acids, and amino acids

DEFF Research Database (Denmark)

Mourtzakis, Marina; Saltin, B.; Graham, T.

2006-01-01

During prolonged exercise, carbohydrate oxidation may result from decreased pyruvate production and increased fatty acid supply and ultimately lead to reduced pyruvate dehydrogenase (PDH) activity. Pyruvate also interacts with the amino acids alanine, glutamine, and glutamate, whereby the decline...... amino acid taken up during exercise and recovery. Alanine and glutamine were also associated...... with pyruvate metabolism, and they comprised 68% of total amino-acid release during exercise and recovery. Thus reduced pyruvate production was primarily associated with reduced carbohydrate oxidation, whereas the greatest production of pyruvate was related to glutamate, glutamine, and alanine metabolism...

Serum free amino acid concentration in hepatic lipidosis of dairy cows in the periparturient period.

Science.gov (United States)

Shibano, Ken-ichi; Kawamura, Seiichi

2006-04-01

Blood samples were taken from eight multiparous cows at a dairy farm on eight occasions between the prepartum period and peak lactation to study the serum concentrations of amino acids and biochemical constituents. The cows were classified as having either severe hepatic lipidosis (HL) or non-hepatic lipidosis (non-HL) according to their clinical condition after calving and changes in serum biochemical parameters. The serum concentrations of beta-hydroxybutyric acid were higher in the HL group than in the non-HL group (ANOVA: phepatic lipidosis.

Specific lysosomal transport of small neutral amino acids

International Nuclear Information System (INIS)

Pisoni, R.L.; Flickinger, K.S.; Thoene, J.G.; Christensen, H.N.

1986-01-01

Studies of amino acid exodus from lysosomes have allowed us previously to describe transport systems specific for cystine and another for cationic amino acids in fibroblast lysosomes. They are now able to study amino acid uptake into highly purified fibroblast lysosomes obtained by separating crude granular fraction on gradients formed by centrifugation in 35% isoosmotic Percoll solutions. Analog inhibition and saturation studies indicate that L-[ 14 C]proline (50 μM) uptake by fibroblast lysosomes at 37 0 C in 50 mM citrate/tris pH 7.0 buffer containing 0.25 M sucrose is mediated by two transport systems, one largely specific for L-proline and the other for which transport is shared with small neutral amino acids such as alanine, serine and threonine. At 7 mM, L-proline inhibits L-[ 14 C]proline uptake almost completely, whereas ala, ser, val, thr, gly, N-methylalanine and sarcosine inhibit proline uptake by 50-65%. The system shared by alanine, serine and threonine is further characterized by these amino acids strongly inhibiting the uptakes of each other. Lysosomal proline transport is selective for the L-isomer of the amino acid, and is scarcely inhibited by 7 mM arg, glu, asp, leu, phe, his, met, (methylamino) isobutyrate, betaine or N,N-dimethylglycine. Cis or trans-4-hydroxy-L-proline inhibit proline uptake only slightly. In sharp contrast to the fibroblast plasma membrane in which Na + is required for most proline and alanine transport, lysosomal uptake of these amino acids occurs independently of Na +

High Serum Essential Amino Acids as a Predictor of Skeletal Muscle Depletion in Patients With Cachexia and Advanced Gastrointestinal Cancers.

Science.gov (United States)

Kitagawa, Moeko; Haji, Seiji; Amagai, Teruyoshi

2017-10-01

In recent years, the number of patients with cancer has increased. These patients are prone to sarcopenia as a result of the decrease in muscle mass and muscle weakness that occur in cancer cachexia. Amino Index Cancer Screening is carried out to evaluate cancer cachexia risk by examining amino acid concentration and analyzing amino acid balance. We conducted a retrospective chart review of consecutive patients with unresectable advanced gastrointestinal cancer (stage IV) receiving chemotherapy treatment (December 2012-September 2015) in an outpatient or in-hospital setting at our institution (N = 46). Data included characteristics, psoas muscle area per computed tomography, and biochemical blood test and serum amino acid profiles. Method 1: Comparison of biomarkers between 2 groups: psoas muscle index change rate (ΔPMI) decrease vs increase. Method 2.1: Correlation between ΔPMI and biomarkers. Method 2.2: Multiple regression of ΔPMI and biomarkers. EAA/TAA ratio (essential amino acids/total amino acids) in the decrease group was significantly higher than that in the increase group. Among all parameters, serum C-reactive protein (CRP), leucine, and isoleucine were negatively related to ΔPMI (correlation coefficients = -0.604, -0.540, -0.518; P = .004, .011, .016, respectively). On multiple regression analysis, serum CRP value was strongly related to ΔPMI ( r 2 = 0.452, β = -0.672, P = .001). Higher serum EAA/TAA ratio and CRP were associated with depletion in psoas muscle area, which led to a diagnosis of sarcopenia, in patients with advanced gastrointestinal cancers. These parameters at baseline could be predictors of cancer cachexia.

Serum Amino Acid Profiles in Normal Subjects and in Patients with or at Risk of Alzheimer Dementia.

Science.gov (United States)

Corso, Gaetano; Cristofano, Adriana; Sapere, Nadia; la Marca, Giancarlo; Angiolillo, Antonella; Vitale, Michela; Fratangelo, Roberto; Lombardi, Teresa; Porcile, Carola; Intrieri, Mariano; Di Costanzo, Alfonso

2017-01-01

Abnormalities in the plasma amino acid profile have been reported in Alzheimer disease (AD), but no data exist for the prodromal phase characterized by subjective memory complaint (SMC). It was our aim to understand if serum amino acid levels change along the continuum from normal to AD, and to identify possible diagnostic biomarkers. Serum levels of 15 amino acids and 2 organic acids were determined in 4 groups of participants - 29 with probable AD, 18 with mild cognitive impairment (MCI), 24 with SMC, and 46 cognitively healthy subjects (HS) - by electrospray tandem mass spectrometry. Glutamate, aspartate, and phenylalanine progressively decreased, while citrulline, argi-ninosuccinate, and homocitrulline progressively increased, from HS over SMC and MCI to AD. The panel including these 6 amino acids and 4 ratios (glutamate/citrulline, citrulline/phenylalanine, leucine plus isoleucine/phenylalanine, and arginine/phenylalanine) discriminated AD from HS with about 96% accuracy. Other panels including 20 biomarkers discriminated SMC or MCI from AD or HS with an accuracy ranging from 88 to 75%. Amino acids contribute to a characteristic metabotype during the progression of AD along the continuum from health to frank dementia, and their monitoring in elderly individuals might help to detect at-risk subjects.

Sequential enzymatic synthesis and separation of 13N-L-glutamic acid and 13N-L-alanine

International Nuclear Information System (INIS)

Cohen, M.B.; Spolter, L.; MacDonald, M.; Chang, C.C.; Takahashi, J.

1975-01-01

The sequential enzymatic synthesis and separation of 13 N-L-glutamic acid and 13 N-L-alanine are described. Basically, that involves the synthesis of 13 N-L-glutamic acid by one enzyme, the transamination of the labeled glutamic acid to form 13 N-L-alanine by a second enzyme, and the separation of the two amino acids by rapid column chromatography. The 13 N-L-alanine was evaluated in animals by imaging and tissue distribution studies and showed good potential as a pancreatic imaging agent

The influence of parenteral nitrogen feeding on free amino acid composition of blood serum and hepatic tissue of irradiated rats

International Nuclear Information System (INIS)

Mil'ko, V.I.; Kirichenko, A.V.; Chalaya, L.A.

1985-01-01

A considerable change in the free am ino acid composition of blood serum and hepatic tissued was noted on the 7th and 14th days following total-body X-irradiation of rats with a dose of 2.9 Gy. The total free amino acid content of blood serum increased and that of hepatic tissue decreased by 85% (on an average) as compared to the intact controls. Quantitative changes in the content of individual amino acids were analysed. Polyamine injected enterally for 7 days and parenterally for 3 days after irradiation a the elimination of the postirradiation changes in the amino acid balance

In Quest of the Alanine R3 Radical: Multivariate EPR Spectral Analyses of X-Irradiated Alanine in the Solid State.

Science.gov (United States)

Jåstad, Eirik O; Torheim, Turid; Villeneuve, Kathleen M; Kvaal, Knut; Hole, Eli O; Sagstuen, Einar; Malinen, Eirik; Futsaether, Cecilia M

2017-09-28

The amino acid l-α-alanine is the most commonly used material for solid-state electron paramagnetic resonance (EPR) dosimetry, due to the formation of highly stable radicals upon irradiation, with yields proportional to the radiation dose. Two major alanine radical components designated R1 and R2 have previously been uniquely characterized from EPR and electron-nuclear double resonance (ENDOR) studies as well as from quantum chemical calculations. There is also convincing experimental evidence of a third minor radical component R3, and a tentative radical structure has been suggested, even though no well-defined spectral signature has been observed experimentally. In the present study, temperature dependent EPR spectra of X-ray irradiated polycrystalline alanine were analyzed using five multivariate methods in further attempts to understand the composite nature of the alanine dosimeter EPR spectrum. Principal component analysis (PCA), maximum likelihood common factor analysis (MLCFA), independent component analysis (ICA), self-modeling mixture analysis (SMA), and multivariate curve resolution (MCR) were used to extract pure radical spectra and their fractional contributions from the experimental EPR spectra. All methods yielded spectral estimates resembling the established R1 spectrum. Furthermore, SMA and MCR consistently predicted both the established R2 spectrum and the shape of the R3 spectrum. The predicted shape of the R3 spectrum corresponded well with the proposed tentative spectrum derived from spectrum simulations. Thus, results from two independent multivariate data analysis techniques strongly support the previous evidence that three radicals are indeed present in irradiated alanine samples.

Serum γ-Glutamyltransferase, Alanine Aminotransferase and Aspartate Aminotransferase Activity in Healthy Blood Donor of Different Ethnic Groups in Gorgan.

Science.gov (United States)

Marjani, Abdoljalal; Mehrpouya, Masoumeh; Pourhashem, Zeinab

2016-07-01

Measure of liver enzymes may help to increase safety of blood donation for both blood donor and recipient. Determination of liver enzymes may prepare valuable clinical information. To assess serum γ-Glutamyltransferase (GGT), Alanine Aminotransferase (ALT), and Aspartate Aminotransferase (AST) activities in healthy blood donors in different ethnic groups in Gorgan. This study was performed in 450 healthy male blood donors, in three ethnic groups (Fars, Sistanee and Turkman) who attended Gorgan blood transfusion center. Liver enzymes (GGT, ALT and AST) were determined. Serum AST and ALT in three ethnic groups were significant except for serum GGT levels. There was significant correlation between family histories of liver disease and systolic blood pressure and AST in Fars, and GGT in Sistanee ethnic groups. Several factors, such as age, family history of diabetes mellitus, family history of liver disease and smoking habit had no effect on some liver enzymes in different ethnic groups in this area. Variation of AST, ALT, and GGT enzyme activities in healthy subjects was associated with some subjects in our study groups. According to our study, it suggests that screening of AST and GGT enzymes in subjects with family history of liver disease is necessary in different ethnic groups.

Metabolism of 3H- and 14C-labeled glutamate, proline, and alanine in normal and adrenalectomized rats using different sites of tracer administration and sampling

International Nuclear Information System (INIS)

Said, H.M.; Chenoweth, M.; Dunn, A.

1989-01-01

Alanine, glutamate and proline labeled with 14C and 3H were infused into fasted normal and adrenalectomized rats. Alanine was administered by the A-V mode (arterial administration-venous sampling), and glutamate and proline by both the A-V and V-A (venous administration-arterial sampling) modes. The kinetics of 14C alanine and 14C glutamate differed markedly from those of the tritium-labeled compounds, but there was little difference in the kinetics of 3H and 14C proline. The replacement rate calculated from the A-V mode for glutamate was about half that obtained in the V-A mode, but there was little difference with proline. The masses of the amino acids (total content of amino acids in the body) were calculated from the washout curves of the tritium-labeled compounds after the infusion of tracer was terminated. The masses for the normal rats were 407 mumol/kg for alanine, 578 mumol/kg for glutamate and 296 mumol/kg for proline. The so-called distribution spaces calculated conventionally from total masses and the amino acid concentrations in plasma are much greater than the volume of the body, reflecting the fact that amino acid concentrations in tissues greatly exceed those in plasma. Adrenalectomy markedly affected the kinetics of the three amino acids, and their replacement rates were greatly reduced. The proline and glutamate masses were reduced by at least one half, while that of alanine was unchanged. Adrenalectomy markedly reduced the conversion of proline to glutamate. The hydrocortisone regimen used in this study restored the metabolism of alanine and glutamate to normal, but had no effect on that of proline

Serum Neuron-Specific Enolase, Biogenic Amino-Acids and Neurobehavioral Function in Lead-Exposed Workers from Lead-Acid Battery Manufacturing Process

OpenAIRE

K Ravibabu; T Barman; HR Rajmohan

2015-01-01

Background: The interaction between serum neuron-specific enolase (NSE), biogenic amino-acids and neurobehavioral function with blood lead levels in workers exposed to lead form lead-acid battery manufacturing process was not studied. Objective: To evaluate serum NSE and biogenic amino-acids (dopamine and serotonin) levels, and neurobehavioral performance among workers exposed to lead from lead-acid storage battery plant, and its relation with blood lead levels (BLLs). Methods: In a c...

Dependence of alanine gel dosimeter response as a function of photon clinical beams dose rate

International Nuclear Information System (INIS)

Silva, Cleber Feijo; Campos, Leticia Lucente

2013-01-01

Gel dosimetry is a new area developed by Gore, it is ery useful for application in radiotherapy because using NMR imaging as evaluation technique is possible to evaluate three dimensional absorbed dose distribution. The measure technique is based on difference of ferrous (Fe 2+ ) and ferric (Fe 3+ ) ) ions concentration that can be measured also by spectrophotometry technique. The Alanine gel dosimeter was developed at IPEN. The alanine is an amino acid and tissue equivalent material that presents significant improvement on previous alanine dosimetry systems. The addition of Alanine increases the production of ferric ions in the solution. This work aims to study the dose rate dependence of photon clinical beams radiation on the alanine gel dosimeter optical response, as well as the response repeatability and gel production reproducibility, since this property is very important for characterization and standardization of any dosimeter. (author)

β-N-Methylamino-L-alanine (BMAA) perturbs alanine, aspartate and glutamate metabolism pathways in human neuroblastoma cells as determined by metabolic profiling.

Science.gov (United States)

Engskog, Mikael K R; Ersson, Lisa; Haglöf, Jakob; Arvidsson, Torbjörn; Pettersson, Curt; Brittebo, Eva

2017-05-01

β-Methylamino-L-alanine (BMAA) is a non-proteinogenic amino acid that induces long-term cognitive deficits, as well as an increased neurodegeneration and intracellular fibril formation in the hippocampus of adult rodents following short-time neonatal exposure and in vervet monkey brain following long-term exposure. It has also been proposed to be involved in the etiology of neurodegenerative disease in humans. The aim of this study was to identify metabolic effects not related to excitotoxicity or oxidative stress in human neuroblastoma SH-SY5Y cells. The effects of BMAA (50, 250, 1000 µM) for 24 h on cells differentiated with retinoic acid were studied. Samples were analyzed using LC-MS and NMR spectroscopy to detect altered intracellular polar metabolites. The analysis performed, followed by multivariate pattern recognition techniques, revealed significant perturbations in protein biosynthesis, amino acid metabolism pathways and citrate cycle. Of specific interest were the BMAA-induced alterations in alanine, aspartate and glutamate metabolism and as well as alterations in various neurotransmitters/neuromodulators such as GABA and taurine. The results indicate that BMAA can interfere with metabolic pathways involved in neurotransmission in human neuroblastoma cells.

Accumulation of D- vs. L-isomers of alanine and leucine in rat prostatic adenocarcinoma

International Nuclear Information System (INIS)

Conti, P.S.; Schmall, B.; Bigler, R.E.; Zanzonico, P.B.; Kleinert, E.; Whitmore, W.F. Jr.

1985-01-01

It has been reported that tumor tissue may accumulate some D-amino acids preferentially over the L-isomers. In order to investigate the potential use of carbon-11 labeled amino acid isomers for in vivo tumor studies with positron emission tomography in patients, the tissue distributions of alanine and leucine, substrates for the A-type and L-type amino acid transport systems, respectively, were studied in Copenhagen rates bearing the Dunning R3327G prostatic adenocarcinoma. The authors have previously reported differences in the accumulation of A-type vs. L-type amino acids in rat prostatic adenocarcinoma and normal tissues. All compounds were labeled with C-14 in the carboxyl position with specific activities of 30.0-56.6 mCi/mmol. Higher levels of C-14 activity (Relative Concentration (RC)=dpm found per gm tissue + dpm inject per gm animal mass) were observed in tumor tissue using D-alanine (0.71) compared to L- (0.21) or DL-alanine (0.27) at 45 min post-injection. While tumor/prostate and tumor/liver ratios were above 2 for all three substrates, tumor/blood and tumor/muscle were above one for only the D-isomer. Comparisons made with D-, L-, and DL-leucine also demonstrated a higher level of RC in tumor tissue with the D-isomer (0.84) vs. the L-(0.66) and DL-leucine (0.63). In this case, however, tumor/blood, tumor/prostate, and tumor/muscle ratios were above one for all three substrates, while tumor/liver ratios were below one. These results support the observation of a preferential accumulation of D-amino acids in tumor tissue over the natural L-isomers. Observed differences in the accumulation of the isomers in normal tissues are discussed

An efficient protocol for the complete incorporation of methyl-protonated alanine in perdeuterated protein

International Nuclear Information System (INIS)

Ayala, Isabel; Sounier, Remy; Use, Nathalie; Gans, Pierre; Boisbouvier, Jerome

2009-01-01

A strategy for the introduction of ( 1 H, 13 C-methyl)-alanine into perdeuterated proteins is described. Specific protonation of alanine methyl groups to a level of 95% can be achieved by overexpressing proteins in M9/D 2 O based bacterial growth medium supplemented with 800 mg/l of 2-[ 2 H], 3-[ 13 C] l-alanine. However, though simple, this approach results in undesired, non-specific background labeling due to isotope scrambling via different amino acid metabolic pathways. Following a careful analysis of known metabolic pathways we found that co-addition of perdeuterated forms of α-ketoisovalerate-d 7 , succinate-d 4 and l-isoleucine-d 10 with labeled l-alanine, reduces undesired background labeling to <1%. When combined with recently developed methyl TROSY experiments, this methyl-specific labeling protocol permits the acquisition of excellent quality correlation spectra of alanine methyl groups in high molecular weight proteins. Our cost effective strategy offers a significant enhancement in the level of incorporation of methyl-labeled alanine in overexpressed proteins over previously reported methods

Importance of intrahepatic mechanisms to gluconeogenesis from alanine during exercise and recovery

International Nuclear Information System (INIS)

Wasserman, D.H.; Williams, P.E.; Lacy, D.B.; Green, D.R.; Cherrington, A.D.

1988-01-01

These studies were performed to assess the importance of intrahepatic mechanisms to gluconeogenesis in the dog during 150 min of treadmill exercise and 90 min of recovery. Sampling catheters were implanted in an artery and portal and hepatic veins 16 days before experimentation. Infusions of [U- 14 C]alanine, [3- 3 H]glucose, and indocyanine green were used to assess gluconeogenesis. During exercise, a decline in arterial and portal vein plasma alanine and in hepatic blood flow led to a decrease in hepatic alanine delivery. During recovery, hepatic blood flow was restored to basal, causing an increase in hepatic alanine delivery beyond exercise rates but still below resting rates. Hepatic fractional alanine extraction increased from 0.26 +/- 0.02 at rest to 0.64 +/- 0.03 during exercise and remained elevated during recovery. Net hepatic alanine uptake was 2.5 +/- 0.2 mumol.kg-1.min-1 at rest and remained unchanged during exercise but was increased during recovery. The conversion rate of [ 14 C]alanine to glucose had increased by 248 +/- 38% by 150 min of exercise and had increased further during recovery. The efficiency with which alanine was channeled into glucose in the liver was accelerated to a rate of 338 +/- 55% above basal by 150 min of exercise but declined slightly during recovery. In conclusion, 1) gluconeogenesis from alanine is accelerated during exercise, due to an increase in the hepatic fractional extraction of the amino acid and through intrahepatic mechanisms that more efficiently channel it into glucose

Role of amino acids and vitamins in nutrition of mesophilic Methanococcus spp

International Nuclear Information System (INIS)

Whitman, W.B.; Sohn, S.; Kuk, S.; Xing, R.

1987-01-01

In this study the authors found that autotrophic methanococci similar to Methanococcus maripaludis obtained up to 57% of their cellular carbon from exogeneous amino acids. About 85% of the incorporation was into protein. Primarily nonpolar and basic amino acids and glycine were incorporated; only small amounts of acidic and some polar amino acids were taken up. An additional 10% of the incorporation was into the nucleic acid fraction. Because little 14 CO 2 was formed from the 14 C-amino acids, little metabolism of the amino acids occurred. Therefore, the growth stimulation by amino acids was probably due to the sparing of anabolic energy requirements. Of the amino acids incorporated, only alanine was also a sole nitrogen source for these methanococci. In contrast, Methanococcus vannielii and Methanococcus aeolicus are autotrophic methanococci which did not incorporate amino acids and did not utilize alanine as a sole nitrogen source. Although glutamine served as a sole nitrogen source for the autotrophic methanococci and Methanococcus voltae, a heterotrophic methanococcus, growth was due to chemical deamination in the medium. M. voltae requires leucine and isoleucine for growth. However, these amino acids were not significant nitrogen sources, and alanine was not a sole nitrogen source for the growth of M. voltae. The branched-chain amino acids were not extensively metabolized by M. voltae. Pantoyl lactone and pantoic acid were readily incorporated by M. voltae. The intact vitamin pantothenate was neither stimulatory to growth nor incorporated. In conclusion, although amino acids and vitamins are nutritionally important to both autotrophic and heterotrophic methanococci, generally they are not subject to extensive catabolism

PGC-1α regulates alanine metabolism in muscle cells.

Science.gov (United States)

Hatazawa, Yukino; Qian, Kun; Gong, Da-Wei; Kamei, Yasutomi

2018-01-01

The skeletal muscle is the largest organ in the human body, depositing energy as protein/amino acids, which are degraded in catabolic conditions such as fasting. Alanine is synthesized and secreted from the skeletal muscle that is used as substrates of gluconeogenesis in the liver. During fasting, the expression of PGC-1α, a transcriptional coactivator of nuclear receptors, is increased in the liver and regulates gluconeogenesis. In the present study, we observed increased mRNA expression of PGC-1α and alanine aminotransferase 2 (ALT2) in the skeletal muscle during fasting. In C2C12 myoblast cells overexpressing PGC-1α, ALT2 expression was increased concomitant with an increased alanine level in the cells and medium. In addition, PGC-1α, along with nuclear receptor ERR, dose-dependently enhanced the ALT2 promoter activity in reporter assay using C2C12 cells. In the absence of glucose in the culture medium, mRNA levels of PGC-1α and ALT2 increased. Endogenous PGC-1α knockdown in C2C12 cells reduced ALT2 gene expression level, induced by the no-glucose medium. Taken together, in the skeletal muscle, PGC-1α activates ALT2 gene expression, and alanine production may play roles in adaptation to fasting.

Dextran Sulfate Sodium Inhibits Alanine Synthesis in Caco-2 Cells

Directory of Open Access Journals (Sweden)

Carolyn M. Slupsky

2011-04-01

Full Text Available To understand and characterize the pathogenic mechanisms of inflammatory bowel disease, dextran sulfate sodium (DSS has been used to induce acute and chronic colitis in animal models by causing intestinal epithelium damage. The mechanism of action of DSS in producing this outcome is not well understood. In an effort to understand how DSS might impact epithelial cell metabolism, we studied the intestinal epithelial cell line Caco-2 incubated with 1% DSS over 56 hours using 1H NMR spectroscopy. We observed no difference in cell viability as compared to control cultures, and an approximately 1.5-fold increase in IL-6 production upon incubation with 1% DSS. The effect on Caco-2 cell metabolism as measured through changes in the concentration of metabolites in the cell supernatant included a three-fold decrease in the concentration of alanine. Given that the concentrations of other amino acids in the cell culture supernatant were not different between treated and control cultures over 56 hours suggest that DSS inhibits alanine synthesis, specifically alanine aminotransferase, without affecting other key metabolic pathways. The importance of alanine aminotransferase in inflammatory bowel disease is discussed.

Structural analysis and mutant growth properties reveal distinctive enzymatic and cellular roles for the three major L-alanine transaminases of Escherichia coli.

Science.gov (United States)

Peña-Soler, Esther; Fernandez, Francisco J; López-Estepa, Miguel; Garces, Fernando; Richardson, Andrew J; Quintana, Juan F; Rudd, Kenneth E; Coll, Miquel; Vega, M Cristina

2014-01-01

In order to maintain proper cellular function, the metabolism of the bacterial microbiota presents several mechanisms oriented to keep a correctly balanced amino acid pool. Central components of these mechanisms are enzymes with alanine transaminase activity, pyridoxal 5'-phosphate-dependent enzymes that interconvert alanine and pyruvate, thereby allowing the precise control of alanine and glutamate concentrations, two of the most abundant amino acids in the cellular amino acid pool. Here we report the 2.11-Å crystal structure of full-length AlaA from the model organism Escherichia coli, a major bacterial alanine aminotransferase, and compare its overall structure and active site composition with detailed atomic models of two other bacterial enzymes capable of catalyzing this reaction in vivo, AlaC and valine-pyruvate aminotransferase (AvtA). Apart from a narrow entry channel to the active site, a feature of this new crystal structure is the role of an active site loop that closes in upon binding of substrate-mimicking molecules, and which has only been previously reported in a plant enzyme. Comparison of the available structures indicates that beyond superficial differences, alanine aminotransferases of diverse phylogenetic origins share a universal reaction mechanism that depends on an array of highly conserved amino acid residues and is similarly regulated by various unrelated motifs. Despite this unifying mechanism and regulation, growth competition experiments demonstrate that AlaA, AlaC and AvtA are not freely exchangeable in vivo, suggesting that their functional repertoire is not completely redundant thus providing an explanation for their independent evolutionary conservation.

Identification of the roles of individual amino acid residues of the helix E of the major antenna of photosystem II (LHCII) by alanine scanning mutagenesis.

Science.gov (United States)

Liu, Cheng; Rao, Yan; Zhang, Lei; Yang, Chunhong

2014-10-01

The functions of the helix E (W97-F105), an amphiphilic lumenal 310 helix of the major antenna of photosystem II (LHCII), are still unidentified. To elucidate the roles of individual amino acid residue of the helix E, alanine scanning mutagenesis has been performed to mutate every residue of this domain to alanine. The influence of every alanine substitution on the structure and function of LHCII has been investigated biochemically and spectroscopically. The results show that all mutations have little impact on the pigment binding and configuration. However, many mutants presented decreased thermo- or photo-stability compared with the wild type, highlighting the significance of this helix to the stability of LHCII. The most critical residue for stability is W97. The mutant W97A yielded very fragile trimeric pigment protein complexes. The structural analysis revealed that the hydrogen bonding and aromatic interactions between W97, F195, F194 and a water molecule contributed greatly to the stability of LHCII. Moreover, Q103A and F105A have been identified to be able to reinforce the tendency of aggregation in vitro. The structural analysis suggested that the enhancement in aggregation formation for Q103A and F105A might be attributed to the changing hydrophobicity of the region. © The Authors 2014. Published by Oxford University Press on behalf of the Japanese Biochemical Society. All rights reserved.

Earthworms accumulate alanine in response to drought.

Science.gov (United States)

Holmstrup, Martin; Slotsbo, Stine; Henriksen, Per G; Bayley, Mark

2016-09-01

Earthworms have ecologically significant functions in tropical and temperate ecosystems and it is therefore important to understand how these animals survive during drought. In order to explore the physiological responses to dry conditions, we simulated a natural drought incident in a laboratory trial exposing worms in slowly drying soil for about one month, and then analyzed the whole-body contents of free amino acids (FAAs). We investigated three species forming estivation chambers when soils dry out (Aporrectodea tuberculata, Aporrectodea icterica and Aporrectodea longa) and one species that does not estivate during drought (Lumbricus rubellus). Worms subjected to drought conditions (alanine that was significantly upregulated in all tested species. Alanine was the most important FAA reaching 250-650μmolg(-1) dry weight in dehydrated Aporrectodea species and 300μmolg(-1) dry weight in L. rubellus. Proline was only weakly upregulated in some species as were a few other FAAs. Species forming estivation chambers (Aporrectodea spp.) did not show a better ability to conserve body water than the non-estivating species (L. rubellus) at the same drought level. These results suggest that the accumulation of alanine is an important adaptive trait in drought tolerance of earthworms in general. Copyright © 2016 Elsevier Inc. All rights reserved.

Characteristics and application of alanine dosimeter 'Aminogray'

International Nuclear Information System (INIS)

Kashiwazaki, Shigeru; Matsuyama, Shigeki; Hatta, Toshimasa; Yagyu, Hideki; Kojima, Takuji; Tanaka, Ryuichi; Morita, Yohsuke.

1988-01-01

Recently, accompanying the progress of nuclear power generation and space development, the evaluation of reliability for the materials and parts used under irradiation has become important. For the evaluation of reliability, the accurate grasp of radiation dose is the prerequisite. In some case, the measurement of cumulative dose in a long period in an actual environment becomes necessary. In this paper, the characteristics and application of a new dosimeter element 'Aminogray' which is suitable to the above requirement are reported. Aminogray is rodshape element made by forming alanine, a kind of amino acid, using a binder polymer, and the alanine content is 70 wt.%, and the polymer is polystyrene. An element of 3 mm diameter and 30 mm length is enclosed in a polystyrene cylinder of 4 mm thickness. This thickness was determined by considering the electronic equilibrium condition in Co-60 gamma-ray irradiation. The principle of the measurement is to determine a dose by measuring the amount of free radicals produced in alanine by radiation using ESR method. The free radicals are extremely stable, and exist for a long period, and the amount of radical production is proportional to absorbed dose. The development, characteristics and application of Aminogray are reported. (K.I.)

Amino acid catabolism and generation of volatiles by lactic acid bacteria.

Science.gov (United States)

Tavaria, F K; Dahl, S; Carballo, F J; Malcata, F X

2002-10-01

Twelve isolates of lactic acid bacteria, belonging to the Lactobacillus, Lactococcus, Leuconostoc, and Enterococcus genera, were previously isolated from 180-d-old Serra da Estrela cheese, a traditional Portuguese cheese manufactured from raw milk and coagulated with a plant rennet. These isolates were subsequently tested for their ability to catabolize free amino acids, when incubated independently with each amino acid in free form or with a mixture thereof. Attempts were made in both situations to correlate the rates of free amino acid uptake with the numbers of viable cells. When incubated individually, leucine, valine, glycine, aspartic acid, serine, threonine, lysine, glutamic acid, and alanine were degraded by all strains considered; arginine tended to build up, probably because of transamination of other amino acids. When incubated together, the degradation of free amino acids by each strain was dependent on pH (with an optimum pH around 6.0). The volatiles detected in ripened Serra da Estrela cheese originated mainly from leucine, phenylalanine, alanine, and valine, whereas in vitro they originated mainly from valine, phenylalanine, serine, leucine, alanine, and threonine. The wild strains tested offer a great potential for flavor generation, which might justify their inclusion in a tentative starter/nonstarter culture for that and similar cheeses.

Effects of β-alanine administration on selected parameters of oxidative stress and phosphoryltransfer network in cerebral cortex and cerebellum of rats

NARCIS (Netherlands)

Gemelli, Tanise; de Andrade, Rodrigo Binkowski; Rojas, Denise Bertin; Bonorino, Nariélle Ferner; Mazzola, Priscila Nicolao; Tortorelli, Lucas Silva; Funchal, Cláudia; Filho, Carlos Severo Dutra; Wannmacher, Clovis Milton Duval

β-Alanine is a β-amino acid derivative of the degradation of pyrimidine uracil and precursor of the oxidative substrate acetyl-coenzyme A (acetyl-CoA). The accumulation of β-alanine occurs in β-alaninemia, an inborn error of metabolism. Patients with β-alaninemia may develop neurological

The distribution of tritium among the amino acids of proteins obtained from mice exposed to tritiated water

International Nuclear Information System (INIS)

Commerford, S.L.; Carsten, A.L.; Cronkite, E.P.

1983-01-01

The distribution of tritium among the amino acids of serum proteins in mice chronically exposed to tritiated water was determined by ion exchange chromatography of the protein hydrolysate. The specific activity of nonexchangeable tritium in these amino acids relative to the specific activity of tritium in the tissue water of mice ranged from 0.04 for phenylalanine and threonine to 1.0 for glycine and alanine. Since tritium from tissue water can enter the nonexchangeable positions of amino acids only as the result of metabolic processing, the relative specific activity of tritium in each amino acid is an indicator of the extent of such processing. The tritium content of tyrosine and all the amino acids required in the diet for survival is quite low, except for histidine, and can be entirely accounted for by transamination or, in the case of methionine, by transmethylation. The tritium content of the other amino acids is too high to result from such minor processing and must reflect primarily the fraction synthesized de novo. The implications of these findings with respect to the radiobiological consequences of a diet containing tritiated proteins are discussed

Acquisition and Assimilation of Nitrogen as Peptide-Bound and D-Enantiomers of Amino Acids by Wheat

Science.gov (United States)

Hill, Paul W.; Quilliam, Richard S.; DeLuca, Thomas H.; Farrar, John; Farrell, Mark; Roberts, Paula; Newsham, Kevin K.; Hopkins, David W.; Bardgett, Richard D.; Jones, David L.

2011-01-01

Nitrogen is a key regulator of primary productivity in many terrestrial ecosystems. Historically, only inorganic N (NH4 + and NO3 -) and L-amino acids have been considered to be important to the N nutrition of terrestrial plants. However, amino acids are also present in soil as small peptides and in D-enantiomeric form. We compared the uptake and assimilation of N as free amino acid and short homopeptide in both L- and D-enantiomeric forms. Sterile roots of wheat (Triticum aestivum L.) plants were exposed to solutions containing either 14C-labelled L-alanine, D-alanine, L-trialanine or D-trialanine at a concentration likely to be found in soil solution (10 µM). Over 5 h, plants took up L-alanine, D-alanine and L-trialanine at rates of 0.9±0.3, 0.3±0.06 and 0.3±0.04 µmol g−1 root DW h−1, respectively. The rate of N uptake as L-trialanine was the same as that as L-alanine. Plants lost ca.60% of amino acid C taken up in respiration, regardless of the enantiomeric form, but more (ca.80%) of the L-trialanine C than amino acid C was respired. When supplied in solutions of mixed N form, N uptake as D-alanine was ca.5-fold faster than as NO3 -, but slower than as L-alanine, L-trialanine and NH4 +. Plants showed a limited capacity to take up D-trialanine (0.04±0.03 µmol g−1 root DW h−1), but did not appear to be able to metabolise it. We conclude that wheat is able to utilise L-peptide and D-amino acid N at rates comparable to those of N forms of acknowledged importance, namely L-amino acids and inorganic N. This is true even when solutes are supplied at realistic soil concentrations and when other forms of N are available. We suggest that it may be necessary to reconsider which forms of soil N are important in the terrestrial N cycle. PMID:21541281

Amino Acid Content of the Gamma Irradiated Cotton Leaf-Worm, Spodoptera Littoralis (Boisd.)

International Nuclear Information System (INIS)

Sobeiha, A.K.; Sallam, H.A.; El-Shall, S.S.A.

2000-01-01

The effects of gamma irradiation on amino acid content of the cotton leaf worm Spodoptera Littoralis was studied.The identified amino acids in the total body tissue of male moths were Theronine, Serine, Glutamic, Glycine, Alanine, Valine, Cystine, Methionine, Isoleucine, Leucine, Tyrosine, Phenylalanine, Lysine, Histidine and Arginine. The irradiation of full grown male pupae with doses 100,200 and 300 Gy decreased the total quantity of amino acids and the amount of most individual amino acids in male moths of P 1 or F 1 generations with some exceptions for Threonine, Alanine, Glycine, Serine, Valine, Cystine and Methionine which were increased.The effect of irradiation on amino acid content of the reproductive system tissues for each male or female were also studied.The results indicated that irradiation decreased the total quantity of amino acid content of both sexes by increasing the dose and males were more radiosensitive than females. Also, irradiation decreased the amount of individual amino acids in both sexes with certain exceptions, e.g. Alanine, Methionine and Tyrosine which increased in the reproductive system of male, and Methionine which increased by more than four times as control.The amino acid content was determined as well in F 1 egg progeny, which was produced from irradiated males Irradiation doses (100, 200 and 300 Gy) decreased the total quantity of amino acids, and all individual ones except Cystine.The greatest reduction (54.9% was observed with Lysine at 300 Gy as compared to control

Branched-chain amino acid supplementation during bed rest: effect on recovery

Science.gov (United States)

Stein, T. P.; Donaldson, M. R.; Leskiw, M. J.; Schluter, M. D.; Baggett, D. W.; Boden, G.

2003-01-01

Bed rest is associated with a loss of protein from the weight-bearing muscle. The objectives of this study are to determine whether increasing dietary branched-chain amino acids (BCAAs) during bed rest improves the anabolic response after bed rest. The study consisted of a 1-day ambulatory period, 14 days of bed rest, and a 4-day recovery period. During bed rest, dietary intake was supplemented with either 30 mmol/day each of glycine, serine, and alanine (group 1) or with 30 mmol/day each of the three BCAAs (group 2). Whole body protein synthesis was determined with U-(15)N-labeled amino acids, muscle, and selected plasma protein synthesis with l-[(2)H(5)]phenylalanine. Total glucose production and gluconeogenesis from alanine were determined with l-[U-(13)C(3)]alanine and [6,6-(2)H(2)]glucose. During bed rest, nitrogen (N) retention was greater with BCAA feeding (56 +/- 6 vs. 26 +/- 12 mg N. kg(-1). day(-1), P < 0.05). There was no effect of BCAA supplementation on either whole body, muscle, or plasma protein synthesis or the rate of 3-MeH excretion. Muscle tissue free amino acid concentrations were increased during bed rest with BCAA (0.214 +/- 0.066 vs. 0.088 +/- 0.12 nmol/mg protein, P < 0.05). Total glucose production and gluconeogenesis from alanine were unchanged with bed rest but were significantly reduced (P < 0.05) with the BCAA group in the recovery phase. In conclusion, the improved N retention during bed rest is due, at least in part, to accretion of amino acids in the tissue free amino acid pools. The amount accreted is not enough to impact protein kinetics in the recovery phase but does improve N retention by providing additional essential amino acids in the early recovery phase.

Effect of Hydroxylamine Sulfate on Volumetric Behavior of Glycine, L-Alanine, and L-Arginine in Aqueous Solution

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Jie Chen

2013-01-01

Full Text Available The apparent molar volumes of glycine, L-alanine, and L-arginine in aqueous hydroxylamine sulfate solutions have been determined at T=298.15 K and atmospheric pressure. The standard partial molar volumes, V20, corresponding partial molar volumes of transfer, ΔtrV20, and hydration numbers, NH, have been calculated for these α-amino acids from the experimental data. The ΔtrV20 values are positive for glycine, L-alanine, and L-arginine and are all increased with the increase in the concentration of hydroxylamine ions. These parameters obtained from the volumetric data are interpreted in terms of various mixing effects between amino acids and hydroxylamine sulfate in aqueous solutions.

A defined medium for Leishmania culture allows definition of essential amino acids.

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Nayak, Archana; Akpunarlieva, Snezhana; Barrett, Michael; Burchmore, Richard

2018-02-01

Axenic culture of Leishmania is generally performed in rich, serum-supplemented media which sustain robust growth over multiple passages. The use of such undefined media, however, obscures proteomic analyses and confounds the study of metabolism. We have established a simple, defined culture medium that supports the sustained growth of promastigotes over multiple passages and which yields parasites that have similar infectivity to macrophages to parasites grown in a conventional semi-defined medium. We have exploited this medium to investigate the amino acid requirements of promastigotes in culture and have found that phenylalanine, tryptophan, arginine, leucine, lysine and valine are essential for viability in culture. Most of the 20 proteogenic amino acids promote growth of Leishmania promastigotes, with the exception of alanine, asparagine, and glycine. This defined medium will be useful for further studies of promastigote substrate requirements, and will facilitate future proteomic and metabolomic analyses. Copyright © 2018 Elsevier Inc. All rights reserved.

On the fragmentation of biomolecules: fragmentation of alanine dipeptide along the polypeptide chain

DEFF Research Database (Denmark)

Solov'yov, Ilia; Yakubovich, Alexander; Solov'yov, Andrey

2006-01-01

The interaction potential between amino acids in alanine dipeptide has been studied for the first time taking into account exact molecular geometry. Ab initio calculation has been performed in the framework of density functional theory taking into account all electrons in the system. The fragment...

Substrate specificity of the aspartate:alanine antiporter (AspT) of Tetragenococcus halophilus in reconstituted liposomes.

Science.gov (United States)

Sasahara, Ayako; Nanatani, Kei; Enomoto, Masaru; Kuwahara, Shigefumi; Abe, Keietsu

2011-08-19

The aspartate:alanine antiporter (AspT) of the lactic acid bacterium Tetragenococcus halophilus is a member of the aspartate:alanine exchanger (AAEx) transporter family. T. halophilus AspT catalyzes the electrogenic exchange of L-aspartate(1-) with L-alanine(0). Although physiological functions of AspT were well studied, L-aspartate(1-):L-alanine(0) antiport mechanisms are still unsolved. Here we report that the binding sites of L-aspartate and L-alanine are independently present in AspT by means of the kinetic studies. We purified His(6)-tagged T. halophilus AspT and characterized its kinetic properties when reconstituted in liposomes (K(m) = 0.35 ± 0.03 mm for L-aspartate, K(m) = 0.098 ± 0 mm for D-aspartate, K(m) = 26 ± 2 mm for L-alanine, K(m) = 3.3 ± 0.2 mm for D-alanine). Competitive inhibition by various amino acids of L-aspartate or L-alanine in self-exchange reactions revealed that L-cysteine selectively inhibited L-aspartate self-exchange but only weakly inhibited L-alanine self-exchange. Additionally, L-serine selectively inhibited L-alanine self-exchange but barely inhibited L-aspartate self-exchange. The aspartate analogs L-cysteine sulfinic acid, L-cysteic acid, and D-cysteic acid competitively and strongly inhibited L-aspartate self-exchange compared with L-alanine self-exchange. Taken together, these kinetic data suggest that the putative binding sites of L-aspartate and L-alanine are independently located in the substrate translocation pathway of AspT.

Substrate Specificity of the Aspartate:Alanine Antiporter (AspT) of Tetragenococcus halophilus in Reconstituted Liposomes*

Science.gov (United States)

Sasahara, Ayako; Nanatani, Kei; Enomoto, Masaru; Kuwahara, Shigefumi; Abe, Keietsu

2011-01-01

The aspartate:alanine antiporter (AspT) of the lactic acid bacterium Tetragenococcus halophilus is a member of the aspartate:alanine exchanger (AAEx) transporter family. T. halophilus AspT catalyzes the electrogenic exchange of l-aspartate1− with l-alanine0. Although physiological functions of AspT were well studied, l-aspartate1−:l-alanine0 antiport mechanisms are still unsolved. Here we report that the binding sites of l-aspartate and l-alanine are independently present in AspT by means of the kinetic studies. We purified His6-tagged T. halophilus AspT and characterized its kinetic properties when reconstituted in liposomes (Km = 0.35 ± 0.03 mm for l-aspartate, Km = 0.098 ± 0 mm for d-aspartate, Km = 26 ± 2 mm for l-alanine, Km = 3.3 ± 0.2 mm for d-alanine). Competitive inhibition by various amino acids of l-aspartate or l-alanine in self-exchange reactions revealed that l-cysteine selectively inhibited l-aspartate self-exchange but only weakly inhibited l-alanine self-exchange. Additionally, l-serine selectively inhibited l-alanine self-exchange but barely inhibited l-aspartate self-exchange. The aspartate analogs l-cysteine sulfinic acid, l-cysteic acid, and d-cysteic acid competitively and strongly inhibited l-aspartate self-exchange compared with l-alanine self-exchange. Taken together, these kinetic data suggest that the putative binding sites of l-aspartate and l-alanine are independently located in the substrate translocation pathway of AspT. PMID:21719707

Taurine and β-alanine intraperitoneal injection in lactating mice modifies the growth and behavior of offspring.

Science.gov (United States)

Nishigawa, Takuma; Nagamachi, Satsuki; Chowdhury, Vishwajit S; Yasuo, Shinobu; Furuse, Mitsuhiro

2018-01-08

Taurine, one of the sulfur-containing amino acids, has several functions in vivo. It has been reported that taurine acts on γ-aminobutyric acid receptors as an agonist and to promote inhibitory neurotransmission. Milk, especially colostrum, contains taurine and it is known that milk taurine is essential for the normal development of offspring. β-Alanine is transported via a taurine transporter and a protein-assisted amino acid transporter, the same ones that transport taurine. The present study aimed to investigate whether the growth and behavior of offspring could be altered by modification of the taurine concentration in milk. Pregnant ICR mice were separated into 3 groups: 1) a control group, 2) a taurine group, and 3) a β-alanine group. During the lactation periods, dams were administered, respectively, with 0.9% saline (10 ml/kg, i.p.), taurine dissolved in 0.9% saline (43 mg/10 ml/kg, i.p.), or β-alanine dissolved in 0.9% saline (31 mg/10 ml/kg, i.p.). Interestingly, the taurine concentration in milk was significantly decreased by the administration of β-alanine, but not altered by the taurine treatment. The body weight of offspring was significantly lower in the β-alanine group. β-Alanine treatment caused a significant decline in taurine concentration in the brains of offspring, and it was negatively correlated with total distance traveled in the open field test at postnatal day 15. Thus, decreased taurine concentration in the brain induced hyperactivity in offspring. These results suggested that milk taurine may have important role of regulating the growth and behavior of offspring. Copyright © 2017 Elsevier Inc. All rights reserved.

Influence of the collection tube on metabolomic changes in serum and plasma.

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López-Bascón, M A; Priego-Capote, F; Peralbo-Molina, A; Calderón-Santiago, M; Luque de Castro, M D

2016-04-01

Major threats in metabolomics clinical research are biases in sampling and preparation of biological samples. Bias in sample collection is a frequently forgotten aspect responsible for uncontrolled errors in metabolomics analysis. There is a great diversity of blood collection tubes for sampling serum or plasma, which are widely used in metabolomics analysis. Most of the existing studies dealing with the influence of blood collection on metabolomics analysis have been restricted to comparison between plasma and serum. However, polymeric gel tubes, which are frequently proposed to accelerate the separation of serum and plasma, have not been studied. In the present research, samples of serum or plasma collected in polymeric gel tubes were compared with those taken in conventional tubes from a metabolomics perspective using an untargeted GC-TOF/MS approach. The main differences between serum and plasma collected in conventional tubes affected to critical pathways such as the citric acid cycle, metabolism of amino acids, fructose and mannose metabolism and that of glycerolipids, and pentose and glucuronate interconversion. On the other hand, the polymeric gel only promoted differences at the metabolite level in serum since no critical differences were observed between plasma collected with EDTA tubes and polymeric gel tubes. Thus, the main changes were attributable to serum collected in gel and affected to the metabolism of amino acids such as alanine, proline and threonine, the glycerolipids metabolism, and two primary metabolites such as aconitic acid and lactic acid. Therefore, these metabolite changes should be taken into account in planning an experimental protocol for metabolomics analysis. Copyright © 2016 Elsevier B.V. All rights reserved.

Inhibition of glutamine synthesis induces glutamate dehydrogenase-dependent ammonia fixation into alanine in co-cultures of astrocytes and neurons.

Science.gov (United States)

Dadsetan, Sherry; Bak, Lasse K; Sørensen, Michael; Keiding, Susanne; Vilstrup, Hendrik; Ott, Peter; Leke, Renata; Schousboe, Arne; Waagepetersen, Helle S

2011-09-01

It has been previously demonstrated that ammonia exposure of neurons and astrocytes in co-culture leads to net synthesis not only of glutamine but also of alanine. The latter process involves the concerted action of glutamate dehydrogenase (GDH) and alanine aminotransferase (ALAT). In the present study it was investigated if the glutamine synthetase (GS) inhibitor methionine sulfoximine (MSO) would enhance alanine synthesis by blocking the GS-dependent ammonia scavenging process. Hence, co-cultures of neurons and astrocytes were incubated for 2.5h with [U-(13)C]glucose to monitor de novo synthesis of alanine and glutamine in the absence and presence of 5.0 mM NH(4)Cl and 10 mM MSO. Ammonia exposure led to increased incorporation of label but not to a significant increase in the amount of these amino acids. However, in the presence of MSO, glutamine synthesis was blocked and synthesis of alanine increased leading to an elevated content intra- as well as extracellularly of this amino acid. Treatment with MSO led to a dramatic decrease in glutamine content and increased the intracellular contents of glutamate and aspartate. The large increase in alanine during exposure to MSO underlines the importance of the GDH and ALAT biosynthetic pathway for ammonia fixation, and it points to the use of a GS inhibitor to ameliorate the brain toxicity and edema induced by hyperammonemia, events likely related to glutamine synthesis. Copyright © 2011 Elsevier Ltd. All rights reserved.

Sorption and Microbial Uptake of Alanine, Glucose and Acetate in Soil

Science.gov (United States)

Fischer, H.; Ingwersen, J.; Kuzyakov, Y.

2009-04-01

Low molecular weight organic substances (LMWOS), e. g. amino acids, sugars, and carboxylic acids, are C compounds that are most rapidly turned-over in the C cycle of soil. Despite of their importance it is still unknown how sorption to the soil matrix affects their turnover in soil solution. The goals of this study were (1) to describe the dynamics of the fluxes of LMWOS (10 µmol l-1) in various pools (dissolved, adsorbed, decomposed to CO2, incorporated into microbial biomass) and (2) to assess the LMWOS distribution in these pools in dependence of very wide range of concentration (0.01 to 1000 µmol l-1). Representatives of each LMWOS group (glucose for sugars, alanine for amino acids, Na-acetate for carboxylic acids) uniformly labeled with 14C were added to sterilized or non-sterilized soil and analyzed in dif-ferent compartments between 1 min and 5.6 hours after addition. LMWOS were almost completely taken up by microorganisms within the first 30 min. Microbial uptake was much faster than the physicochemical sorption (estimated in sterilized soil), which needed to reach quasi-equilibrium 60 min for alanine and about 400 min for glucose. Only sorption of acetate was instantaneous (>1 min). While for acetate the maximum sorption capacity was reached at 100 µmol l-1 no such maximum was found for glucose and alanine in the studied concentra-tion range. At the concentration of 100 µmol l-1, microbial decomposition after 4.5 h hours was higher for alanine (76.7±1.1%) than acetate (55.2±0.9%) and glucose (28.5±1.5%). On the contrary, incorporation into microbial biomass was higher for glucose (59.8±1.2%) than for acetate (23.4±5.9%) and alanine (5.2±2.8%). Within 10 to 500 µmol l-1 the pathways of the three LMWOS transformation changed: at 500 µmol l-1 alanine and acetate were less mineralized and more incorporated into microbial biomass than at 10 µmol l-1, while glucose incorporation decreased. Consequently, the concentrations of alanine, glucose, and

Amino Acid Signatures to Evaluate the Beneficial Effects of Weight Loss

DEFF Research Database (Denmark)

Geidenstam, Nina; Magnusson, Martin; Danielsson, Anders P H

2017-01-01

-associated amino acids from the MDC-CC, alanine, isoleucine, tyrosine, phenylalanine, and glutamate decreased after weight loss, while asparagine increased after weight maintenance. These changes were validated in the replication cohort. Scores that were constructed based on obesity-associated amino acids...

Post-translational Introduction of D-Alanine into Ribosomally Synthesized Peptides by the Dehydroalanine Reductase NpnJ.

Science.gov (United States)

Yang, Xiao; van der Donk, Wilfred A

2015-10-07

Ribosomally synthesized peptides are generally limited to L-amino acid building blocks. Given the advantageous properties of peptides containing D-amino acids such as stabilization of certain turns and against proteolytic degradation, methods to introduce D-stereocenters are valuable. Here we report the first in vitro reconstitution and characterization of a dehydrogenase that carries out the asymmetric reduction of dehydroalanine. NpnJA reduces dehydroalanine to D-Ala using NAPDH as cosubstrate. The enzyme displays high substrate tolerance allowing introduction of D-Ala into a range of non-native substrates. In addition to the in vitro reactions, we describe five examples of using Escherichia coli as biosynthetic host for D-alanine introduction into ribosomal peptides. A deuterium-label-based coupled-enzyme assay was used to rapidly determine the stereochemistry of the newly installed alanine.

High serum alanine aminotransferase is associated with the risk of colorectal adenoma in Korean men.

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Moon, Chang Mo; Yun, Kyung Eun; Ryu, Seungho; Chang, Yoosoo; Park, Dong Il

2017-07-01

An elevated alanine aminotransferase (ALT) is frequently observed in subjects with metabolic syndrome, which is associated with the risk of colorectal adenoma (CRA). However, the relationship between ALT and CRA remains unclear. Therefore, we aimed to investigate whether high serum ALT is associated with the risk of CRA in a metabolically healthy population. We conducted this cross-sectional study in 27,717 asymptomatic Korean adults who underwent a health checkup. Subjects were categorized as adenoma-free, hyperplastic polyp, low-risk adenoma, or high-risk adenoma. High-risk adenoma was defined as three or more adenomas, any adenoma ≥ 10 mm, or adenoma with high-grade dysplasia or villous features. Among all participants, 10.3% and 1.5% of cases were categorized as low-risk and high-risk adenoma, respectively. In multivariate analyses adjusting for age, sex, body mass index, smoking habits, alcohol intake, regular exercise, aspirin and analgesics use, family history of colon cancer, education level, fatty liver, high-sensitivity C reactive protein, homeostasis model assessment of insulin resistance, total cholesterol, and triglyceride, an increase in ALT was positively associated with the prevalence of low-risk and high-risk adenoma (P for trend = 0.029 and 0.027, respectively). The highest quartile group of ALT level showed a significantly increased prevalence in low-risk (odds ratio, 1.17) and high-risk adenoma (odds ratio, 1.48) groups compared with the lowest quartile group. This phenomenon persisted in the subgroup analysis in men, but not in women. In the asymptomatic healthy population, high serum ALT is significantly associated with the risk of CRA. © 2016 Journal of Gastroenterology and Hepatology Foundation and John Wiley & Sons Australia, Ltd.

Effects of Glutamine and Alanine Supplementation on Central Fatigue Markers in Rats Submitted to Resistance Training

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Audrey Yule Coqueiro

2018-01-01

Full Text Available Recent evidence suggests that increased brain serotonin synthesis impairs performance in high-intensity intermittent exercise and specific amino acids may modulate this condition, delaying fatigue. This study investigated the effects of glutamine and alanine supplementation on central fatigue markers in rats submitted to resistance training (RT. Wistar rats were distributed in: sedentary (SED, trained (CON, trained and supplemented with alanine (ALA, glutamine and alanine in their free form (G + A, or as dipeptide (DIP. Trained groups underwent a ladder-climbing exercise for eight weeks, with progressive loads. In the last 21 days, supplementations were offered in water with a 4% concentration. Albeit without statistically significance difference, RT decreased liver glycogen, and enhanced the concentrations of plasma glucose, free fatty acids (FFA, hypothalamic serotonin, and ammonia in muscle and the liver. Amino acids affected fatigue parameters depending on the supplementation form. G + A prevented the muscle ammonia increase by RT, whereas ALA and DIP augmented ammonia and glycogen concentrations in muscle. DIP also increased liver ammonia. ALA and G + A reduced plasma FFA, whereas DIP increased this parameter, free tryptophan/total tryptophan ratio, hypothalamic serotonin, and the serotonin/dopamine ratio. The supplementations did not affect physical performance. In conclusion, glutamine and alanine may improve or impair central fatigue markers depending on their supplementation form.

Persistent GABAA/C responses to gabazine, taurine and beta-alanine in rat hypoglossal motoneurons.

Science.gov (United States)

Chesnoy-Marchais, D

2016-08-25

In hypoglossal motoneurons, a sustained anionic current, sensitive to a blocker of ρ-containing GABA receptors, (1,2,5,6-tetrahydropyridin-4-yl)methylphosphinic acid (TPMPA) and insensitive to bicuculline, was previously shown to be activated by gabazine. In order to better characterize the receptors involved, the sensitivity of this atypical response to pentobarbital (30μM), allopregnanolone (0.3μM) and midazolam (0.5μM) was first investigated. Pentobarbital potentiated the response, whereas the steroid and the benzodiazepine were ineffective. The results indicate the involvement of hybrid heteromeric receptors, including at least a GABA receptor ρ subunit and a γ subunit, accounting for the pentobarbital-sensitivity. The effects of the endogenous β amino acids, taurine and β-alanine, which are released under various pathological conditions and show neuroprotective properties, were then studied. In the presence of the glycine receptor blocker strychnine (1μM), both taurine (0.3-1mM) and β-alanine (0.3mM) activated sustained anionic currents, which were partly blocked by TPMPA (100μM). Thus, both β amino acids activated ρ-containing GABA receptors in hypoglossal motoneurons. Bicuculline (20μM) reduced responses to taurine and β-alanine, but small sustained responses persisted in the presence of both strychnine and bicuculline. Responses to β-alanine were slightly increased by allopregnanolone, indicating a contribution of the bicuculline- and neurosteroid-sensitive GABAA receptors underlying tonic inhibition in these motoneurons. Since sustained activation of anionic channels inhibits most mature principal neurons, the ρ-containing GABA receptors permanently activated by taurine and β-alanine might contribute to some of their neuroprotective properties under damaging overexcitatory situations. Copyright © 2016 IBRO. Published by Elsevier Ltd. All rights reserved.

Chiral separation of amino acids in biological fluids by micellar electrokinetic chromatography with laser-induced fluorescence detection.

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Thorsén, G; Bergquist, J

2000-08-18

A method is presented for the chiral analysis of amino acids in biological fluids using micellar electrokinetic chromatography (MEKC) and laser-induced fluorescence (LIF). The amino acids are derivatized with the chiral reagent (+/-)-1-(9-anthryl)-2-propyl chloroformate (APOC) and separated using a mixed micellar separation system. No tedious pre-purification of samples is required. The excellent separation efficiency and good detection capabilities of the MEKC-LIF system are exemplified in the analysis of urine and cerebrospinal fluid. This is the first time MEKC has been reported for chiral analysis of amino acids in biological fluids. The amino acids D-alanine, D-glutamine, and D-aspartic acid have been observed in cerebrospinal fluid, and D-alanine and D-glutamic acid in urine. To the best of our knowledge no measurements of either D-alanine in cerebrospinal fluid or D-glutamic acid in urine have been presented in the literature before.

Influence of different yeasts on the amino acid pattern of rosé wine

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Mandl Karin

2017-01-01

Full Text Available In an experiment with Rosé wine, 27 different commercial yeasts were tested for their influence on the amino acid pattern of the wine. Amino acids are precursors for aromatic substances; therefore a large variation of the amino acid values in the wine was expected. Blaufränkisch grapes with 20° KMW were matured in the cellar with 27 different commercial yeasts. The fermentation was carried out in 34l vessels. The wines were measured for amino acids using an HP 1200 liquid chromatograph and HP-FLD1100 according to Umagat. The wines showed 13.5% alcohol and little residual sugar. The measurement results of the amino acids of the different wines showed large variations. For example, the amount of the amino acid alanine in wine varied from 17 to 138 mg. In particular, the wines of the yeast Pino Type showed the highest amounts of alanine in comparison to the other fermented wines.

Serum Alanine Aminotransferase Levels within Normal Range Have Different Associations with Augmentation Index and Other Cardiometabolic Risk Factors in Nondrinkers and Drinkers: A Chinese Community-Based Analysis

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Shihui Fu

2017-01-01

Full Text Available Background. To investigate whether serum alanine aminotransferase (ALT levels within normal range were associated with augmentation index (AIx and cardiometabolic risk factors in nondrinkers and drinkers in Chinese community-dwelling population. Methods. There were 4165 participants with serum ALT levels within normal range. Results. Alcohol drinking was observed in 1173 participants (28.2%. In multivariate analysis, serum ALT levels of nondrinkers were independently associated with age, sex, body mass index (BMI, hypertension, diabetes mellitus, diastolic blood pressure, triglyceride, low-density lipoprotein-cholesterol (LDL-c, and AIx, while serum ALT levels of drinkers were independently associated with age, sex, BMI, triglyceride, and LDL-c (p<0.05 for all. Conclusions. Associations of serum ALT levels within normal range with age, sex, body height and weight, and blood lipid were simultaneously present in participants with and without alcohol drinking, while associations of serum ALT levels within normal range with AIx, blood pressure, and glucose were seen in nondrinkers rather than in drinkers. These findings not only provide the evidence that serum ALT levels, even within the normal range, have different associations with arteriosclerosis and cardiometabolic risk factors in nondrinkers and drinkers but also are helpful in understanding the underlying pathophysiologic mechanisms linking the hepatic function to arteriosclerosis and cardiometabolic risk factors.

Dose response of alanine and methyl alanine towards gamma and in-situ alpha irradiation

International Nuclear Information System (INIS)

Mohapatra, M.; Rajeswari, B.; Bhide, M.K.; Rane, Vinayak; Kadam, R.M.

2017-01-01

In situ alpha and external gamma dose response of two ESR (electron spin resonance) dosimetric materials namely alanine and methyl alanine were investigated. It was observed that alanine dosimeter had a better dose response in comparison to methyl alanine for the in-situ alpha irradiation by using 239 Pu powder. On the other hand, in case of gamma radiation, methyl alanine was found to have the sensitivity as twice that of alanine. (author)

Synthesis of 15N isotope labeled alanine

International Nuclear Information System (INIS)

Oliveira, Claudineia R. de; Bendassolli, Jose Albertino; Sant'Ana, Carlos Roberto; Tagliassachi, Romulo Barbieri; Maximo, Everaldo; Prestes, Clelber Vieira

2005-01-01

The application of light chemical elements and their stable isotopes in biological studies have been increased over the last years. The use of 15 N labeled amino acids is an important tool for elucidation of peptides structures. This paper describe a method for the synthesis of 15 N isotope labeled alanine at lower costs than international ones, as well as the details of the recovery system of the nitrogen residues. In the present work an amination of α-haloacids, with the bromopropionic carboxylic acid and labeled aqua ammonia ( 15 NH 3 aq) was carried out. In order to avoid eventually losses of 15 NH 3 , special cares were adopted, since the production cost is high. Although the acquisition cost of the 13 N (radioactive) labeled compounds is lower, the obtained stable tracer will allow the accomplishment of important studies of the nitrogen cycling in living things, less occupational and environment hazards, and the time limitation problems in field studies. The tests took place in triplicates with NH 3 (aq) being employed. With the establishment of the system for 15 NH 3 recovery, an average of 94 % of the ammonia employed in the synthesis process was recovered. The purity of the amino acid was state determined by TLC (Thin Layer Chromatography) and HPLC (High-Performance Liquid Chromatography) with a fluorescence detector. The Rf and the retention time of the synthesized sample were similar the sigma standard. Finally, regarding the established conditions, it was possible to obtain the alanine with a production cost about 40 % lower than the international price. (author)

Effects of high-salinity seawater acclimation on the levels of D-alanine in the muscle and hepatopancreas of kuruma prawn, Marsupenaeus japonicus.

Science.gov (United States)

Yoshikawa, Naoko; Yokoyama, Masahumi

2015-12-10

Changes in D- and L-alanine contents were determined in the muscle and hepatopancreas of kuruma prawn Marsupenaeus japonicus, during acclimation from seawater containing 100% salinity to artificial seawater containing 150% salinity. In the hepatopancreas, contents of both amino acids increased by approximately threefold. The activity of alanine racemase, which catalyzes the interconversion of D- and L-alanine, also increased in the high-salinity seawater. In addition, the expression of the gene encoding alanine racemase increased in the hepatopancreas with an increase in the alanine racemase activity. These data indicate that the biosynthesis of D- and L-alanine is controlled by the gene expression level of alanine racemase, and D-alanine in the hepatopancreas functions as a major osmolyte for isosmotic regulation. In contrast, the content of D-alanine and alanine racemase activity did not change in the muscle during hyper-osmotic acclimation. Therefore, we suggest that D-alanine, which exists in the several tissues of M. japonicus, is considered to be utilized in some different physiological phenomena in different tissues. Copyright © 2015 Elsevier B.V. All rights reserved.

DEUTERIUM FRACTIONATION DURING AMINO ACID FORMATION BY PHOTOLYSIS OF INTERSTELLAR ICE ANALOGS CONTAINING DEUTERATED METHANOL

Energy Technology Data Exchange (ETDEWEB)

Oba, Yasuhiro; Watanabe, Naoki; Kouchi, Akira [Institute of Low Temperature Science, Hokkaido University, N19W8, Kita-ku, Sapporo, Hokkaido 060-0819 (Japan); Takano, Yoshinori, E-mail: oba@lowtem.hokudai.ac.jp [Department of Biogeochemistry, Japan Agency for Marine-Earth Science and Technology (JAMSTEC), 2-15 Natsushima, Yokosuka, Kanagawa 237-0061 (Japan)

2016-08-10

Deuterium (D) atoms in interstellar deuterated methanol might be distributed into complex organic molecules through molecular evolution by photochemical reactions in interstellar grains. In this study, we use a state-of-the-art high-resolution mass spectrometer coupled with a high-performance liquid chromatography system to quantitatively analyze amino acids and their deuterated isotopologues formed by the photolysis of interstellar ice analogs containing singly deuterated methanol CH{sub 2}DOH at 10 K. Five amino acids (glycine, α -alanine, β -alanine, sarcosine, and serine) and their deuterated isotopologues whose D atoms are bound to carbon atoms are detected in organic residues formed by photolysis followed by warming up to room temperature. The abundances of singly deuterated amino acids are in the range of 0.3–1.1 relative to each nondeuterated counterpart, and the relative abundances of doubly and triply deuterated species decrease with an increasing number of D atoms in a molecule. The abundances of amino acids increase by a factor of more than five upon the hydrolysis of the organic residues, leading to decreases in the relative abundances of deuterated species for α -alanine and β -alanine. On the other hand, the relative abundances of the deuterated isotopologues of the other three amino acids did not decrease upon hydrolysis, indicating different formation mechanisms of these two groups upon hydrolysis. The present study facilitates both qualitative and quantitative evaluations of D fractionation during molecular evolution in the interstellar medium.

DEUTERIUM FRACTIONATION DURING AMINO ACID FORMATION BY PHOTOLYSIS OF INTERSTELLAR ICE ANALOGS CONTAINING DEUTERATED METHANOL

International Nuclear Information System (INIS)

Oba, Yasuhiro; Watanabe, Naoki; Kouchi, Akira; Takano, Yoshinori

2016-01-01

Deuterium (D) atoms in interstellar deuterated methanol might be distributed into complex organic molecules through molecular evolution by photochemical reactions in interstellar grains. In this study, we use a state-of-the-art high-resolution mass spectrometer coupled with a high-performance liquid chromatography system to quantitatively analyze amino acids and their deuterated isotopologues formed by the photolysis of interstellar ice analogs containing singly deuterated methanol CH 2 DOH at 10 K. Five amino acids (glycine, α -alanine, β -alanine, sarcosine, and serine) and their deuterated isotopologues whose D atoms are bound to carbon atoms are detected in organic residues formed by photolysis followed by warming up to room temperature. The abundances of singly deuterated amino acids are in the range of 0.3–1.1 relative to each nondeuterated counterpart, and the relative abundances of doubly and triply deuterated species decrease with an increasing number of D atoms in a molecule. The abundances of amino acids increase by a factor of more than five upon the hydrolysis of the organic residues, leading to decreases in the relative abundances of deuterated species for α -alanine and β -alanine. On the other hand, the relative abundances of the deuterated isotopologues of the other three amino acids did not decrease upon hydrolysis, indicating different formation mechanisms of these two groups upon hydrolysis. The present study facilitates both qualitative and quantitative evaluations of D fractionation during molecular evolution in the interstellar medium.

Production of a new D-amino acid oxidase from the fungus Fusarium oxysporum.

Science.gov (United States)

Gabler, M; Fischer, L

1999-08-01

The fungus Fusarium oxysporum produced a D-amino acid oxidase (EC 1. 4.3.3) in a medium containing glucose as the carbon and energy source and ammonium sulfate as the nitrogen source. The specific D-amino acid oxidase activity was increased up to 12.5-fold with various D-amino acids or their corresponding derivatives as inducers. The best inducers were D-alanine (2.7 microkat/g of dry biomass) and D-3-aminobutyric acid (2.6 microkat/g of dry biomass). The addition of zinc ions was necessary to permit the induction of peroxisomal D-amino acid oxidase. Bioreactor cultivations were performed on a 50-liter scale, yielding a volumetric D-amino acid oxidase activity of 17 microkat liter(-1) with D-alanine as an inducer. Under oxygen limitation, the volumetric activity was increased threefold to 54 microkat liter(-1) (3,240 U liter(-1)).

Radiation-induced reactions in D, L-α-alanine adsorbed in solid surfaces

International Nuclear Information System (INIS)

Aguilar, E; Negrón-Mendoza, A.; Camargo, C.

2013-01-01

The aim of this work is to study the behavior under irradiation of D, L and D-L α-alanine adsorbed in solid surfaces, as possible phase in the chemical evolution that may have occurred on the primitive Earth or in extraterrestrial environments and to evaluate the contribution of solids (a clay mineral) as shields for the adsorbed amino acids against a external energy source. The results show that α-alanine is adsorbed in the surfaces as function of pH and its yield of decomposition in mineral suspension is lower than the system without the solid surface. These results show the importance of nuclear techniques in these types of studies. (author)

Kinetic enantioselectivity of a protonated bis(diamido)-bridged basket resorcin[4]arene towards alanine peptides.

Science.gov (United States)

Fraschetti, C; Montagna, M; Crestoni, M E; Calcaterra, A; Aiello, F; Santi, L; Filippi, A

2017-02-01

Efficient enantiodiscrimination of some alanine-containing di- and tri-peptides by using chiral protonated bis(diamido)-bridged basket resorcin[4]arenes depends on several factors, including the basicity of the amino acid residues at the C- and N-termini of the peptide.

Protein Homeostasis Defects of Alanine-Glyoxylate Aminotransferase: New Therapeutic Strategies in Primary Hyperoxaluria Type I

Directory of Open Access Journals (Sweden)

Angel L. Pey

2013-01-01

Full Text Available Alanine-glyoxylate aminotransferase catalyzes the transamination between L-alanine and glyoxylate to produce pyruvate and glycine using pyridoxal 5′-phosphate (PLP as cofactor. Human alanine-glyoxylate aminotransferase is a peroxisomal enzyme expressed in the hepatocytes, the main site of glyoxylate detoxification. Its deficit causes primary hyperoxaluria type I, a rare but severe inborn error of metabolism. Single amino acid changes are the main type of mutation causing this disease, and considerable effort has been dedicated to the understanding of the molecular consequences of such missense mutations. In this review, we summarize the role of protein homeostasis in the basic mechanisms of primary hyperoxaluria. Intrinsic physicochemical properties of polypeptide chains such as thermodynamic stability, folding, unfolding, and misfolding rates as well as the interaction of different folding states with protein homeostasis networks are essential to understand this disease. The view presented has important implications for the development of new therapeutic strategies based on targeting specific elements of alanine-glyoxylate aminotransferase homeostasis.

Markerless deletion of putative alanine dehydrogenase genes in Bacillus licheniformis using a codBA-based counterselection technique.

Science.gov (United States)

Kostner, David; Rachinger, Michael; Liebl, Wolfgang; Ehrenreich, Armin

2017-11-01

Bacillus licheniformis strains are used for the large-scale production of industrial exoenzymes from proteinaceous substrates, but details of the amino acid metabolism involved are largely unknown. In this study, two chromosomal genes putatively involved in amino acid metabolism of B. licheniformis were deleted to clarify their role. For this, a convenient counterselection system for markerless in-frame deletions was developed for B. licheniformis. A deletion plasmid containing up- and downstream DNA segments of the chromosomal deletion target was conjugated to B. licheniformis and integrated into the genome by homologous recombination. Thereafter, the counterselection was done by using a codBA cassette. The presence of cytosine deaminase and cytosine permease exerted a conditionally lethal phenotype on B. licheniformis cells in the presence of the cytosine analogue 5-fluorocytosine. Thereby clones were selected that lost the integrated vector sequence and the anticipated deletion target after a second recombination step. This method allows the construction of markerless mutants in Bacillus strains in iterative cycles. B. licheniformis MW3 derivatives lacking either one of the ORFs BL03009 or BL00190, encoding a putative alanine dehydrogenase and a similar putative enzyme, respectively, retained the ability to grow in minimal medium supplemented with alanine as the carbon source. In the double deletion mutant MW3 ΔBL03009 ΔBL00190, however, growth on alanine was completely abolished. These data indicate that the two encoded enzymes are paralogues fulfilling mutually replaceable functions in alanine utilization, and suggest that in B. licheniformis MW3 alanine utilization is initiated by direct oxidative transamination to pyruvate and ammonium.

Uptake of neutral alpha- and beta-amino acids by human proximal tubular cells

DEFF Research Database (Denmark)

Jessen, H; Røigaard, H; Jacobsen, Christian

1996-01-01

experiments revealed that all the neutral amino acids tested reduced the uptake of AIB, whereas there was no effect of taurine, L-aspartic acid, and L-arginine. By contrast, the influx of beta-alanine was only drastically reduced by beta-amino acids, whereas the inhibition by neutral alpha-amino acids...

Radiolysis of alanine adsorbed in a clay mineral

Energy Technology Data Exchange (ETDEWEB)

Aguilar-Ovando, Ellen Y.; Negron-Mendoza, Alicia [Instituto de Ciencias Nucleares, Universidad Nacional Autonoma de Mexico (UNAM), Circuito Exterior s/n, Ciudad Universitaria, Apartado Postal 70-543, Deleg. Coyoacan, C.P. 04510 (Mexico)

2013-07-03

Optical activity in molecules is a chemical characteristic of living beings. In this work, we examine the hypothesis of the influence of different mineral surfaces on the development of a specific chirality in organic molecules when subjected to conditions simulating the primitive Earth during the period of chemical evolution. By using X-ray diffraction techniques and HPLC/ELSD to analyze aqueous suspensions of amino acids adsorbed on minerals irradiated in different doses with a cobalt-60 gamma source, the experiments attempt to prove the hypothesis that some solid surfaces (like clays and meteorite rocks) may have a concentration capacity and protective role against external sources of ionizing radiation (specifically {gamma}-ray) for some organic compounds (like some amino acids) adsorbed on them. Preliminary results show a slight difference in the adsorption and radiolysis of the D-and L-alanine.

Radiolysis of alanine adsorbed in a clay mineral

International Nuclear Information System (INIS)

Aguilar-Ovando, Ellen Y.; Negrón-Mendoza, Alicia

2013-01-01

Optical activity in molecules is a chemical characteristic of living beings. In this work, we examine the hypothesis of the influence of different mineral surfaces on the development of a specific chirality in organic molecules when subjected to conditions simulating the primitive Earth during the period of chemical evolution. By using X-ray diffraction techniques and HPLC/ELSD to analyze aqueous suspensions of amino acids adsorbed on minerals irradiated in different doses with a cobalt-60 gamma source, the experiments attempt to prove the hypothesis that some solid surfaces (like clays and meteorite rocks) may have a concentration capacity and protective role against external sources of ionizing radiation (specifically γ-ray) for some organic compounds (like some amino acids) adsorbed on them. Preliminary results show a slight difference in the adsorption and radiolysis of the D-and L-alanine

Amino acid profile of raw and boiled seeds of african walnut ...

African Journals Online (AJOL)

Principal Component Analysis (PCA) explained close to 50% of the total variability in amino acid composition, identifying arginine, asparagine, lysine, methionine, valine, glutamic acid, leucine, cysteine, threonine, alanine and isoleucine as the key amino acids for describing African walnut seeds in the south-eastern zone of ...

Effects of clustering structure on volumetric properties of amino acids in (DMSO + water) mixtures

International Nuclear Information System (INIS)

Huang Aimin; Liu Chunli; Ma Lin; Tong Zhangfa; Lin Ruisen

2012-01-01

Graphical abstract: Together with static light scattering measurement, volumetric properties of glycine, L-alanine and L-serine were determined and utilized to reveal the microscopic solvent structure of (DMSO + water) mixtures and its influence on the interaction between DMSO and amino acids from a clustering point of view. The results demonstrated that the interaction between amino acids and DMSO was greatly related to the clustering structure of the mixed solvent and that amino acids interacted with already established solvent clusters. Hydrophobic aggregating of DMSO lead to a decrease in the hydrophobic effect of DMSO and the hydrophobic–hydrophilic and hydrophobic–hydrophobic interaction with amino acids, which was reflected by the solvation of proteins. Highlights: ► Determine volumetric properties of three amino acids in aqueous DMSO in details. ► Static light scattering measurement for clustering structure of aqueous DMSO. ► Volumetric behaviour of amino acids depends on clustering structure of aqueous DMSO. ► Clustering structure of aqueous DMSO influences solvation of protein and cellulose. - Abstract: For a better understanding on the functions of DMSO in biological systems at a relatively lower concentration, apparent molar volumes of three typical amino acids, glycine, L-alanine and L-serine in (DMSO + water) mixtures were determined and the transfer volumes from water to the mixtures were evaluated. Together with static light scattering measurement, the results were utilised to reveal the microscopic solvent structure of (DMSO + water) mixtures and its influence on the interaction between DMSO and amino acids from a clustering point of view. The results demonstrate that the interaction between amino acids and DMSO is greatly related to the clustering structure of the mixed solvent and that amino acids interacted with already established solvent clusters. The linear dependence of transfer volume of amino acids on DMSO concentration up to 2

Radical formation of irradiated α-alanine and N-acetyl alanine with heavy ion beams. Effects of the irradiation temperature

International Nuclear Information System (INIS)

Minegishi, Atsuko; Nagasaki, Jun; Mori, Wasuke; Amano, Chikara; Takagi, Shinji; Murakami, Takeshi; Kanai, Tatsuaki; Furusawa, Yoshiya; Iwata, Yoshiyuki

2003-01-01

The characteristics of irradiation with C290 MeV/u ion beams were investigated using X-band electron spin resonance (ESR) spectroscopy for a polycrystalline powder of L-α-alanine at from 77K to 310K. The formed main radicals at 190K∼310K were the deamino radical and the decarboxyl radical. Because of the first-derivative ESR, decarboxyl radical showed an expanded spectral width and a lower peak height because of its amino hydrogen and nitrogen than that of the same amount of deamino radical. The ESR of irradiated L-α-alanine predominantly indicates the spectrum of the deamino radical. On the irradiated, L-α-alanine at from 77K to 310K ESR showed 1:4:6:4:1 lines at 220K and at room temperature, which indicate that the methyl group of the radical was rotating. On the other hand, at 77K ESR the spectrum showed nearly 1:5:5:5:1 lines, like the teeth of a saw, on samples irradiated at 270K∼350K (range IV), and 1:4:6:4:1 lines for those irradiated at 180K∼260K (range II and III), respectively. It is considered that the radical conformation of the deamino radical is planar (most stable conformation) on an irradiated sample in range IV, and a pyramidal structure on the irradiated sample in ranges II and III. (author)

Amino acids and hexosamines as indicators of organic matter degradation state in North Sea sediments

NARCIS (Netherlands)

Dauwe, B.; Middelburg, J.J.

1998-01-01

Sediment cores from six stations in the eastern North Sea were analyzed for protein amino acids, the nonprotein amino acids beta-alanine and gamma-aminobutyric acid and the hexosamines galactosamine and glucosamine, and bulk parameters (organic carbon, nitrogen, total hydrolyzable amino acids and

Amino acids in biofilm material on aluminium panels immersed in marine waters

Digital Repository Service at National Institute of Oceanography (India)

Bhosle, N.B.; Wagh, A.B.

.63 to 52.04% of the total ON of the SPM. Aspartic acid, glycine, alanine, serine, leucine and lysine and glutamic acid were the most abundant individual amino acids in both the biofilm material and the SPM. THe distribution of individual amino acids...

Glutamine and alanine-induced differential expression of intracellular IL-6, IL-8, and TNF-α in LPS-stimulated monocytes in human whole-blood.

Science.gov (United States)

Raspé, C; Czeslick, E; Weimann, A; Schinke, C; Leimert, A; Kellner, P; Simm, A; Bucher, M; Sablotzki, A

2013-04-01

To investigate the effects of the commonly-used immunomodulators l-glutamine, l-alanine, and the combination of both l-alanyl-l-glutamine (Dipeptamin(®)) on intracellular expression of IL-6, IL-8, and TNF-α during endotoxemia, lipopolysaccharide (LPS)-stimulated human monocytes in a whole blood system were investigated by flow cytometry. Whole blood of twenty-seven healthy volunteers was stimulated with LPS and incubated with three different amino acid solutions (1. l-glutamine, 2. l-alanine, 3. l-alanyl-l-glutamine, each concentration 2 mM, 5 mM, incubation time 3 h). CD14(+) monocytes were phenotyped in whole-blood and intracellular expression of cytokines was assessed by flow cytometry. Our investigations showed for the first time in whole blood probes, imitating best physiologically present cellular interactions, that l-glutamine caused a dose-independent inhibitory effect on IL-6 and TNF-α production in human monocytes stimulated with LPS. However, l-alanine had contrary effects on IL-6 expression, significantly upregulating expression of IL-6 in LPS-treated monocytes. The impact of l-alanine on the expression of TNF-α was comparable with glutamine. Neither amino acid was able to affect IL-8 production in LPS-stimulated monocytes. The combination of both did not influence significantly IL-6 and IL-8 expression in monocytes during endotoxemia, however strongly reduced TNF-α production. For the regulation of TNF-α, l-glutamine, l-alanine and the combination of both show a congruent and exponentiated downregulating effect during endotoxemia, for the modulation of IL-6, l-glutamine and l-alanine featured opposite regulation leading to a canceling impact of each other when recombining both amino acids. Copyright © 2013 Elsevier Ltd. All rights reserved.

Preferential Treatment: Interaction Between Amino Acids and Minerals

Science.gov (United States)

Crapster-Pregont, E. J.; Cleaves, H. J.; Hazen, R. M.

2008-12-01

Amino acids are the building blocks of proteins and are important for some models of the origin of life. Polymerization of amino acids from dilute solution is unlikely without a scaffold or catalyst. The surfaces of early Earth minerals are the most likely candidates for this role. The surface adsorption behavior of 12 amino acids (L-alanine, L-serine, L-aspartic acid, L-proline, L- phenylalanine, L-valine, L-arginine, d-amino valeric acid, glycine, L-lysine, L-isoleucine, and B-alanine) on 21 minerals (quartz, calcite, enstatite, illite, olivine, pyrrhotite, pyrite, alkali basalt, albite, analcime, chlorite, barite, hydroxyl apatite, hematite, magnetite, aluminum hydroxide, kaolin, silica gel, corundum, rutile, and montmorillonite) was determined via batch adsorption experiments. Absorption was determined for concentrations between 10-4M and 10-6M in the presence of 0.1M NaCl, and between pH values of 3 and 9 at 25 degrees C. The equilibrated solutions were centrifuged, filtered, derivatized using a fluorescent amino group tag (dansyl-chloride) and analyzed by HPLC. Adsorption was standardized using BET surface area measurements for each mineral to give the number of mols of each amino acid adsorbed per square meter for each mineral. The results indicate an enormous difference in the adsorption of amino acids between minerals, along with major differences in the adsorption of individual amino acids on the same mineral surface. There is also a change in the absorbance of amino acids as the pH changes. Many previous studies of amino acid concentration and catalysis by minerals have used clay minerals because of their high surface areas, however, this data suggests that the surfaces of minerals such as calcite, quartz and pyrite have even higher affinities for amino acids. The results suggest mineral surfaces that could be optimal locations for the polymerization of molecules linked to the origin of life.

Studies on radiolysis of amino acids, 1

International Nuclear Information System (INIS)

Oku, Tadatake

1977-01-01

In order to elucidate the radiolysis of amino acid, peptide, protein and enzyme, the radiolytic mechanisms of neutral amino acids (glycine, L-alanine, L-valine, L-leucine, L-isoleucine, L-serine, and L-threonine) and acidic amino acids (L-aspartic acid, L-glutamic acid and DL-amino-n-adipic acid) were studied in the presence of air or in the atmosphere nitrogen. An aqueous solution of 1 mM. of each amino acid was sealed in a glass ampoule under air or nitrogen. Irradiation of amino acid solutions was carried out with γ-rays of 60 Co at doses of 4.4-2,640x10 3 rads. The amino acids and the radiolytic products formed were determined by ion-exchange chromatography. From the results of determining amino acids and the radiolytic products formed and their G-values, the radiolytic mechanisms of the amino acids were discussed. (auth.)

A non-enzymic browning induced by gamma cobalt-60 irradiation and heating in a fructose-alanine model system

International Nuclear Information System (INIS)

Bachman, S.; Zegota, A.; Zegota, H.

1981-01-01

The Maillard browning reaction between reducing sugars and amino compounds is important in food chemistry and may considerably affect the colour, aroma and nutritional value of food after thermal processing. In this study, the effect of irradiation combined with heating on the course of browning reaction in the model system of aqueous solution of fructose (0.03M) and alanine (0.01M) was investigated. The optical absorption spectra recorded for irradiated and heated solution of fructose-alanine were different from those of only irradiated or only heated solution. The brown colour of the samples is caused by the extension of the tail-end absorption into the visible region of the spectrum. No absorption maximum appears in the visible range. The heating of irradiated fructose solution with non-irradiated alanine develops markedly more intensive browning than that of the heating of irradiated alanine solution with non-irradiated fructose. The products of fructose radiolysis are responsible for the acceleration of browning in the fructose-alanine system. (author)

Branched-chain amino acid supplementation during bed rest: effect on recovery

Science.gov (United States)

Stein, T. P.; Donaldson, M. R.; Leskiw, M. J.; Schluter, M. D.; Baggett, D. W.; Boden, G.

2003-01-01

Bed rest is associated with a loss of protein from the weight-bearing muscle. The objectives of this study are to determine whether increasing dietary branched-chain amino acids (BCAAs) during bed rest improves the anabolic response after bed rest. The study consisted of a 1-day ambulatory period, 14 days of bed rest, and a 4-day recovery period. During bed rest, dietary intake was supplemented with either 30 mmol/day each of glycine, serine, and alanine (group 1) or with 30 mmol/day each of the three BCAAs (group 2). Whole body protein synthesis was determined with U-(15)N-labeled amino acids, muscle, and selected plasma protein synthesis with l-[(2)H(5)]phenylalanine. Total glucose production and gluconeogenesis from alanine were determined with l-[U-(13)C(3)]alanine and [6,6-(2)H(2)]glucose. During bed rest, nitrogen (N) retention was greater with BCAA feeding (56 +/- 6 vs. 26 +/- 12 mg N. kg(-1). day(-1), P BCAA supplementation on either whole body, muscle, or plasma protein synthesis or the rate of 3-MeH excretion. Muscle tissue free amino acid concentrations were increased during bed rest with BCAA (0.214 +/- 0.066 vs. 0.088 +/- 0.12 nmol/mg protein, P BCAA group in the recovery phase. In conclusion, the improved N retention during bed rest is due, at least in part, to accretion of amino acids in the tissue free amino acid pools. The amount accreted is not enough to impact protein kinetics in the recovery phase but does improve N retention by providing additional essential amino acids in the early recovery phase.

Transglycosylated Starch Improves Insulin Response and Alters Lipid and Amino Acid Metabolome in a Growing Pig Model

Directory of Open Access Journals (Sweden)

Monica A. Newman

2017-03-01

Full Text Available Due to the functional properties and physiological effects often associated with chemically modified starches, significant interest lies in their development for incorporation in processed foods. This study investigated the effect of transglycosylated cornstarch (TGS on blood glucose, insulin, and serum metabolome in the pre- and postprandial phase in growing pigs. Eight jugular vein-catheterized barrows were fed two diets containing 72% purified starch (waxy cornstarch (CON or TGS. A meal tolerance test (MTT was performed with serial blood sampling for glucose, insulin, lipids, and metabolome profiling. TGS-fed pigs had reduced postprandial insulin (p < 0.05 and glucose (p < 0.10 peaks compared to CON-fed pigs. The MTT showed increased (p < 0.05 serum urea with TGS-fed pigs compared to CON, indicative of increased protein catabolism. Metabolome profiling showed reduced (p < 0.05 amino acids such as alanine and glutamine with TGS, suggesting increased gluconeogenesis compared to CON, probably due to a reduction in available glucose. Of all metabolites affected by dietary treatment, alkyl-acyl-phosphatidylcholines and sphingomyelins were generally increased (p < 0.05 preprandially, whereas diacyl-phosphatidylcholines and lysophosphatidylcholines were decreased (p < 0.05 postprandially in TGS-fed pigs compared to CON. In conclusion, TGS led to changes in postprandial insulin and glucose metabolism, which may have caused the alterations in serum amino acid and phospholipid metabolome profiles.

Na--dependent transport of basic, zwitterionic, and bicyclic amino acids by a broad-scope system in mouse blastocysts

International Nuclear Information System (INIS)

Van Winkle, L.J.; Christensen, H.N.; Campione, A.L.

1985-01-01

Mouse blastocysts which had been activated from diapause in utero appeared to take up amino acids via a Na - -dependent transport system with novel characteristics. In contrast to other cell types, uptake of 3-aminoendobicyclo [3,2,1]octane-3-carboxylic acid (BCO) by blastocysts was largely Na - dependent. Moreover, L-alanine and BCO met standard criteria for mutual competitive inhibition of the Na - -dependent transport of each other. The Ki for each of these amino acids as an inhibitor of transport of the other had a value similar to the value of its Km for transport. In addition, both 2-aminoendobicyclo [2,2,1]heptane-2-carboxylic acid and L-valine appeared to inhibit Na - -dependent transport of alanine and BCO competitively. Finally, alanine and L-lysine appeared to compete for the same Na+-dependent transport sites in blastocysts. For these reasons, the authors conclude that lysine, alanine, and BCO are transported by a common Na+-dependent system in blastocysts. In addition, the apparent interaction of the system with other basic amino acids, such as 1-dimethylpiperidine-4-amino-4-carboxylic acid, which has a nondissociable positive charge on its side chain, and L-arginine and L-homoarginine, whose cationic forms are highly predominant at neutral pH, suggests that the cationic forms of basic amino acids are transported by the wide-scope system

Alanine dosimetry for clinical applications. Proceedings

International Nuclear Information System (INIS)

Anton, M.

2006-05-01

The following topics are dealt with: Therapy level alanine dosimetry at the UK Nationational Physical Laboratory, alanine as a precision validation tool for reference dosimetry, composition of alanine pellet dosimeters, the angular dependence of the alanine ESR spectrum, the CIAE alanine dosimeter for radiotherapy level, a correction for temporal evolution effects in alanine dosimetry, next-generation services foe e-traceability to ionization radiation national standards, establishing e-traceability to HIST high-dose measurement standards, alanine dosimetry of dose delivery from clinical accelerators, the e-scan alanine dosimeter reader, alanine dosimetry at ISS, verification of the integral delivered dose for IMRT treatment in the head and neck region with ESR/alanine dosimetry, alanine dosimetry in helical tomotherapy beams, ESR dosimetry research and development at the University of Palermo, lithium formate as a low-dose EPR radiation dosimeter, sensitivity enhancement of alanine/EPR dosimetry. (HSI)

Determination of muscle protein synthesis rates in fish using (2)H2O and (2)H NMR analysis of alanine.

Science.gov (United States)

Marques, Cátia; Viegas, Filipa; Rito, João; Jones, John; Viegas, Ivan

2016-09-15

Following administration of deuterated water ((2)H2O), the fractional synthetic rate (FSR) of a given endogenous protein can be estimated by (2)H-enrichment quantification of its alanine residues. Currently, this is measured by mass spectrometry following a derivatization procedure. Muscle FSR was measured by (1)H/(2)H NMR analysis of alanine from seabass kept for 6 days in 5% (2)H-enriched saltwater, following acid hydrolysis and amino acid isolation by cation-exchange chromatography of muscle tissue. The analysis is simple and robust, and provides precise measurements of excess alanine (2)H-enrichment in the 0.1-0.4% range from 50 mmol of alanine recovered from muscle protein. Copyright © 2016 Elsevier Inc. All rights reserved.

Catalytic amino acid production from biomass-derived intermediates

KAUST Repository

Deng, Weiping

2018-04-30

Amino acids are the building blocks for protein biosynthesis and find use in myriad industrial applications including in food for humans, in animal feed, and as precursors for bio-based plastics, among others. However, the development of efficient chemical methods to convert abundant and renewable feedstocks into amino acids has been largely unsuccessful to date. To that end, here we report a heterogeneous catalyst that directly transforms lignocellulosic biomass-derived α-hydroxyl acids into α-amino acids, including alanine, leucine, valine, aspartic acid, and phenylalanine in high yields. The reaction follows a dehydrogenation-reductive amination pathway, with dehydrogenation as the rate-determining step. Ruthenium nanoparticles supported on carbon nanotubes (Ru/CNT) exhibit exceptional efficiency compared with catalysts based on other metals, due to the unique, reversible enhancement effect of NH3 on Ru in dehydrogenation. Based on the catalytic system, a two-step chemical process was designed to convert glucose into alanine in 43% yield, comparable with the well-established microbial cultivation process, and therefore, the present strategy enables a route for the production of amino acids from renewable feedstocks. Moreover, a conceptual process design employing membrane distillation to facilitate product purification is proposed and validated. Overall, this study offers a rapid and potentially more efficient chemical method to produce amino acids from woody biomass components.

Comparison of the Tastes of L-Alanine and Monosodium Glutamate in C57BL/6J Wild Type and T1r3 Knockout Mice.

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Eddy, Meghan C; Eschle, Benjamin K; Delay, Eugene R

2017-09-01

Previous research showed that L-alanine and monosodium L-glutamate elicit similar taste sensations in rats. This study reports the results of behavioral experiments designed to compare the taste capacity of C57BL/6J wild type and T1r3- mice for these 2 amino acids. In conditioned taste aversion (CTA) experiments, wild-type mice exhibited greater sensitivity than knockout mice for both L-amino acids, although knockout mice were clearly able to detect both amino acids at 50 mM and higher concentrations. Generalization of CTA between L-alanine and L-glutamate was bidirectionally equivalent for both mouse genotypes, indicating that both substances elicited similar tastes in both genotypes. This was verified by the discrimination experiments in which both mouse genotypes performed at or near chance levels at 75 and 150 mM. Above 150 mM, discrimination performance improved, suggesting the taste qualities of the 2 L-amino acids are not identical. No differences between knockout and wild-type mice in discrimination ability were detected. These results indicate that while the T1r3 receptor is important for tasting L-alanine and L-glutamate, other receptors are also important for tasting these amino acids. © The Author 2017. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Characterization of Lactobacillus salivarius alanine racemase: short-chain carboxylate-activation and the role of A131.

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Kobayashi, Jyumpei; Yukimoto, Jotaro; Shimizu, Yasuhiro; Ohmori, Taketo; Suzuki, Hirokazu; Doi, Katsumi; Ohshima, Toshihisa

2015-01-01

Many strains of lactic acid bacteria produce high concentrations of d-amino acids. Among them, Lactobacillus salivarius UCC 118 produces d-alanine at a relative concentration much greater than 50 % of the total d, l-alanine (100d/d, l-alanine). We characterized the L. salivarius alanine racemase (ALR) likely responsible for this d-alanine production and found that the enzyme was activated by carboxylates, which is an unique characteristic among ALRs. In addition, alignment of the amino acid sequences of several ALRs revealed that A131 of L. salivarius ALR is likely involved in the activation. To confirm that finding, an L. salivarius ALR variant with an A131K (ALR(A131K)) substitution was prepared, and its properties were compared with those of ALR. The activity of ALR(A131K) was about three times greater than that of ALR. In addition, whereas L. salivarius ALR was strongly activated by low concentrations (e.g., 1 mM) of short chain carboxylates, and was inhibited at higher concentrations (e.g., 10 mM), ALR(A131K) was clearly inhibited at all carboxylate concentrations tested (1-40 mM). Acetate also increased the stability of ALR such that maximum activity was observed at 35 °C and pH 8.0 without acetate, but at 50 °C in the presence of 1 mM acetate. On the other hand, maximum ALR(A131K) activity was observed at 45 °C and around pH 9.0 with or without acetate. It thus appears that A131 mediates the activation and stabilization of L. salivarius ALR by short chain carboxylates.

Dosimetric evaluation of spectrophotometric response of alanine gel solution for gamma, photons, electrons and thermal neutrons radiations

International Nuclear Information System (INIS)

Silva, Cleber Feijo

2009-01-01

Alanine Gel Dosimeter is a new gel material developed at IPEN that presents significant improvement on Alanine system developed by Costa. The DL-Alanine (C 3 H 7 NO 2 ) is an amino acid tissue equivalent that improves the production of ferric ions in the solution. This work aims to analyse the main dosimetric characteristics this new gel material for future application to measure dose distribution. The performance of Alanine gel solution was evaluated to gamma, photons, electrons and thermal neutrons radiations using the spectrophotometry technique. According to the obtained results for the different studied radiation types, the reproducibility intra-batches and inter-batches is better than 4% and 5%, respectively. The dose response presents a linear behavior in the studied dose range. The response dependence as a function of dose rate and incident energy is better 2% and 3%, respectively. The lower detectable dose is 0.1 Gy. The obtained results indicate that the Alanine gel dosimeter presents good performance and can be useful as an alternative dosimeter in the radiotherapy area, using MRI technique for tridimensional dose distribution evaluation. (author)

[Dynamics of the amino acid composition of the medium during cultivation of isolated organs by the directed perfusion method].

Science.gov (United States)

Barashkov, V A; Gitel'zon, I I; Nefedov, V P; Trubachev, I N

1975-07-01

The dynamics of the amino acid composition of the medium under conditions of adequate perfusion of the isolated organs of a dog (sternum, kidney and liver) was studied. It was found that after a 6-hour perfusion of the complex of organs the amount in the perfusion medium of such amino acids as histidine, lysine, alanine, considerably increased, whereas the amount of arginine, serine, aspartic acid, threonine with glutamine, isoleucine, proline, leucine and valine decreased as compared with their initial concentration. The dynamics of the amino acid medium composition during a 4-hour perfusion was studied in experiments with the isolated sternum. The concentration of alanine, lysine and histidine increased in the medium. At the same time there was seen a decrease in the concentration of serine, aspartic acid, isoleucine, tyrosine and phenyl-alanine.

Comparative study on free amino acid composition of wild edible mushroom species.

Science.gov (United States)

Ribeiro, Bárbara; Andrade, Paula B; Silva, Branca M; Baptista, Paula; Seabra, Rosa M; Valentão, Patrícia

2008-11-26

A comparative study on the amino acid composition of 11 wild edible mushroom species (Suillus bellini, Suillus luteus, Suillus granulatus, Tricholomopsis rutilans, Hygrophorus agathosmus, Amanita rubescens, Russula cyanoxantha, Boletus edulis, Tricholoma equestre, Fistulina hepatica, and Cantharellus cibarius) was developed. To define the qualitative and quantitative profiles, a derivatization procedure with dabsyl chloride was performed, followed by HPLC-UV-vis analysis. Twenty free amino acids (aspartic acid, glutamic acid, asparagine, glutamine, serine, threonine, glycine, alanine, valine, proline, arginine, isoleucine, leucine, tryptophan, phenylalanine, cysteine, ornithine, lysine, histidine, and tyrosine) were determined. B. edulis and T. equestre were revealed to be the most nutritional species, whereas F. hepatica was the poorest. The different species exhibited distinct free amino acid profiles. The quantification of the identified compounds indicated that, in a general way, alanine was the major amino acid. The results show that the analyzed mushroom species possess moderate amino acid contents, which may be relevant from a nutritional point of view because these compounds are indispensable for human health. A combination of different mushroom species in the diet would offer good amounts of amino acids and a great diversity of palatable sensations.

Amino Acid Chemistry as a Link Between Small Solar System Bodies and Carbonaceous Chondrites

Science.gov (United States)

Glavin, Daniel P.; Ehrenfreund, Pascale; Botta, Oliver; Cooper, George; Bada, Jeffrey L.

2000-01-01

Establishing chemical links between meteorites and small solar system bodies, such as comets and asteroids, provides a tool for investigating the processes that occurred during the formation of the solar system. Carbonaceous meteorites are of particular interest, since they may have seeded the early Earth with a variety of prebiotic organic compounds including amino acids, purines and pyrimidines, which are thought to be necessary for the origin of life. Here we report the results of high-performance liquid chromatography (HPLC) based amino acid analyses of the acid-hydrolyzed hot water extracts from pristine interior pieces of the CI carbonaceous chondrites Orgueil and Ivuna and the CM meteorites Murchison and Murray. We found that the CI meteorites Orgueil and Ivuna contained high abundances of beta-alanine and glycine, while only traces of other amino acids like alanine, alpha-amino-n-butryic acid (ABA) and alpha-aminoisobutyric acid (AIB) were detected in these meteorites. Carbon isotopic measurements of beta-alanine and glycine in Orgueil by gas chromatography combustion-isotope ratio mass spectrometry clearly indicate an extraterrestrial origin of these amino acids. The amino acid composition of Orgueil and Ivuna was strikingly different from the CM chondrites Murchison and Murray. The most notable difference was the high relative abundance of B-alanine in Orgueil and Ivuna compared to Murchison and Murray. Furthermore, AIB, which is one of the most abundant amino acids found in Murchison and Murray, was present in only trace amounts in Orgueil and Ivuna. Our amino acid data strongly suggest that the CI meteorites Orgueil and Ivuna came from a different type of parent body than the CM meteorites Murchison and Murray, possibly from an extinct comet. It is generally thought that carbonaceous meteorites are fragments of larger asteroidal bodies delivered via near Earth objects (NEO). Orbital and dynamic studies suggest that both fragments of main belt asteroids

An allostatic mechanism for M2 pyruvate kinase as an amino-acid sensor.

Science.gov (United States)

Yuan, Meng; McNae, Iain W; Chen, Yiyuan; Blackburn, Elizabeth A; Wear, Martin A; Michels, Paul A M; Fothergill-Gilmore, Linda A; Hupp, Ted; Walkinshaw, Malcolm D

2018-05-10

We have tested the effect of all 20 proteinogenic amino acids on the activity of the M2 isoenzyme of pyruvate kinase (M2PYK) and show that within physiologically relevant concentrations, phenylalanine, alanine, tryptophan, methionine, valine, and proline act as inhibitors while histidine and serine act as activators. Size exclusion chromatography has been used to show that all amino acids, whether activators or inhibitors, stabilise the tetrameric form of M2PYK. In the absence of amino-acid ligands an apparent tetramer-monomer dissociation K d is estimated to be ~0.9 µM with a slow dissociation rate (t 1/2 ~ 15 min). X-ray structures of M2PYK complexes with alanine, phenylalanine, and tryptophan show the M2PYK locked in an inactive T-state conformation, while activators lock the M2PYK tetramer in the active R-state conformation. Amino-acid binding in the allosteric pocket triggers rigid body rotations (11°) stabilising either T or R-states. The opposing inhibitory and activating effects of the non-essential amino acids serine and alanine suggest that M2PYK could act as a rapid-response nutrient sensor to rebalance cellular metabolism. This competition at a single allosteric site between activators and inhibitors provides a novel regulatory mechanism by which M2PYK activity is finely tuned by the relative (but not absolute) concentrations of activator and inhibitor amino acids. Such 'allostatic' regulation may be important in metabolic reprogramming and influencing cell fate. ©2018 The Author(s).

Altered tryptophan and alanine transport in fibroblasts from boys with attention-deficit/hyperactivity disorder (ADHD: an in vitro study

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Vumma Ravi

2011-09-01

Full Text Available Abstract Background The catecholaminergic and serotonergic neurotransmitter systems are implicated in the pathophysiology of attention-deficit/hyperactivity disorder (ADHD. The amino acid tyrosine is the precursor for synthesis of the catecholamines dopamine and norepinephrine, while tryptophan is the precursor of serotonin. A disturbed transport of tyrosine, as well as other amino acids, has been found in a number of other psychiatric disorders, such as schizophrenia, bipolar disorder and autism, when using the fibroblast cell model. Hence, the aim of this study was to explore whether children with ADHD may have disturbed amino acid transport. Methods Fibroblast cells were cultured from skin biopsies obtained from 14 boys diagnosed with ADHD and from 13 matching boys without a diagnosis of a developmental disorder. Transport of the amino acids tyrosine, tryptophan and alanine across the cell membrane was measured by the cluster tray method. The kinetic parameters, maximal transport capacity (Vmax and affinity constant (Km were determined. Any difference between the two groups was analyzed by Student's unpaired t-test or the Mann Whitney U test. Results The ADHD group had significantly decreased Vmax (p = 0.039 and Km (increased affinity (p = 0.010 of tryptophan transport in comparison to controls. They also had a significantly higher Vmaxof alanine transport (p = 0.031, but the Km of alanine transport did not differ significantly. There were no significant differences in any of the kinetic parameters regarding tyrosine transport in fibroblasts for the ADHD group. Conclusions Tryptophan uses the same transport systems in both fibroblasts and at the blood brain barrier (BBB. Hence, a decreased transport capacity of tryptophan implies that less tryptophan is being transported across the BBB in the ADHD group. This could lead to deficient serotonin access in the brain that might cause disturbances in both the serotonergic and the catecholaminergic

Higher Ratio of Serum Alpha-Fetoprotein Could Predict Outcomes in Patients with Hepatitis B Virus-Associated Hepatocellular Carcinoma and Normal Alanine Aminotransferase.

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Young-Il Kim

Full Text Available The role of serum alpha-fetoprotein (AFP levels in the surveillance and diagnosis of hepatocellular carcinoma (HCC is controversial. The aim of this study was to investigate the value of serially measured serum AFP levels in HCC progression or recurrence after initial treatment.A total of 722 consecutive patients newly diagnosed with HCC and treated at the National Cancer Center, Korea, between January 2004 and December 2009 were enrolled. The AFP ratios between 4-8 weeks post-treatment and those at the time of HCC progression or recurrence were obtained. Multivariate logistic regression analysis was performed to correlate the post-treatment AFP ratios with the presence of HCC progression or recurrence.The etiology of HCC was related to chronic hepatitis B virus (HBV infection in 562 patients (77.8%, chronic hepatitis C virus (HCV infection in 74 (10.2%, and non-viral cause in 86 (11.9%. There was a significant decrease in serum AFP levels from the baseline to 4 to 8 weeks after treatment (median AFP, 319.6 ng/mL vs. 49.6 ng/mL; p 1.0 was an independently associated with HCC progression or recurrence. Among the different causes of HCC analyzed, this association was significant only for HCC related to chronic hepatitis B (p< 0.001 and non-viral causes (p<0.05, and limited only to patients who had normal alanine aminotransferase (ALT levels.Serial measurements of serum AFP ratios could be helpful in detecting progression or recurrence in treated patients with HBV-HCC and normal ALT.

Higher Ratio of Serum Alpha-Fetoprotein Could Predict Outcomes in Patients with Hepatitis B Virus-Associated Hepatocellular Carcinoma and Normal Alanine Aminotransferase

Science.gov (United States)

Park, Joong-Won

2016-01-01

Background The role of serum alpha-fetoprotein (AFP) levels in the surveillance and diagnosis of hepatocellular carcinoma (HCC) is controversial. The aim of this study was to investigate the value of serially measured serum AFP levels in HCC progression or recurrence after initial treatment. Methods A total of 722 consecutive patients newly diagnosed with HCC and treated at the National Cancer Center, Korea, between January 2004 and December 2009 were enrolled. The AFP ratios between 4–8 weeks post-treatment and those at the time of HCC progression or recurrence were obtained. Multivariate logistic regression analysis was performed to correlate the post-treatment AFP ratios with the presence of HCC progression or recurrence. Results The etiology of HCC was related to chronic hepatitis B virus (HBV) infection in 562 patients (77.8%), chronic hepatitis C virus (HCV) infection in 74 (10.2%), and non-viral cause in 86 (11.9%). There was a significant decrease in serum AFP levels from the baseline to 4 to 8 weeks after treatment (median AFP, 319.6 ng/mL vs. 49.6 ng/mL; p 1.0 was an independently associated with HCC progression or recurrence. Among the different causes of HCC analyzed, this association was significant only for HCC related to chronic hepatitis B (p< 0.001) and non-viral causes (p<0.05), and limited only to patients who had normal alanine aminotransferase (ALT) levels. Conclusion Serial measurements of serum AFP ratios could be helpful in detecting progression or recurrence in treated patients with HBV-HCC and normal ALT. PMID:27304617

Amino acids grafting of Ar+ ions modified PE

International Nuclear Information System (INIS)

Svorcik, V.; Hnatowicz, V.; Stopka, P.; Bacakova, L.; Heitz, J.; Oechsner, R.; Ryssel, H.

2001-01-01

Polyethylene (PE) was irradiated with 63 keV Ar + ions to the fluences from 1x10 12 to 3x10 15 cm -2 and then grafted at room temperature from water solution with amino acids (alanine, leucine). Using various spectroscopic techniques (UV-VIS, FTIR, RBS and EPR) it was shown that the amino acids penetrate into PE where they are eventually captured either on double bonds or on free radicals created by the ion irradiation. Grafting with amino acids in the whole specimen layer modified by irradiation is observed. The ion-beam-modified and amino-acid grafted PE is supposed to exhibit increased biocompatibility. (author)

Pharmacological Identification of a Guanidine-Containing β-Alanine Analogue with Low Micromolar Potency and Selectivity for the Betaine/GABA Transporter 1 (BGT1)

DEFF Research Database (Denmark)

Al-Khawaja, Anas Mohammad Ali; Petersen, Jette Gellert; Damgaard, Maria

2014-01-01

of the amino group in β-alanine or GABA, a series of compounds was generated, and their pharmacological activity assessed at human GAT subtypes. Using a cell-based [(3)H]GABA uptake assay, several selective inhibitors at human BGT1 were identified. The guanidine-containing compound 9 (2-amino-1......,4,5,6-tetrahydropyrimidine-5-carboxylic acid hydrochloride) displayed more than 250 times greater potency than the parent compound β-alanine at BGT1 and is thus the most potent inhibitor reported to date for this subtype (IC50 value of 2.5 µM). In addition, compound 9 displayed about 400, 16 and 40 times lower inhibitory...

A quest for normal values of serum aminotransferase and its association with age and body mass index

International Nuclear Information System (INIS)

Farooq, M.A.; Farooq, Y.; Khan, M.A.

2016-01-01

Objective: To determine the reference range for serum alanine aminotransferase (ALT) levels for young healthy individuals in our population and to find out effects of age and body mass index on alanine aminotransferase. Study Design: Descriptive cross sectional study. Place and Duration of Study: Military Hospital Rawalpindi; from Dec 2009 to April 2010. Material and Methods: Five hundred and fifty three young healthy adults were included in the study. Detailed history was taken. Parameters like Age, height and weight were measured. Relevant investigations were carried out for each subject which included blood complete picture, Chest X-Ray, ECG, Urine routine examination, liver function tests, serum urea, serum creatinine, serum uric acid, plasma glucose (F), Anti HCV antibodies, HBsAg, HIV antibodies and VDRL. Data was analyzed using SPSS version 22. Results: Age of subjects ranged from 19 to 50 years (29 +- 6) years, BMI 15.59 to 29.8, (21.58 +- 2). Serum alanine aminotransferase ranged from 17 to 81 IU/l, (29.9 +- 8) IU/l. Serum alanine aminotransferase for subjects with body mass index more than 25 Kg/m2 was 29.6 +- 7 IU/L. There is no significant difference in value of mean ALT overall and in those with BMI of more than 25 (p=0.149) Conclusion: Mean serum alanine aminotransferase value in young healthy adults included in this study was 29.9 +- 8 IU/L which is lower than normal reference provided at present which is 40 IU/l. Statistical analysis revealed that serum ALT is not significantly influenced by age and BMI. (author)

Role of the gerA operon in L-alanine germination of Bacillus licheniformis spores

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Løvdal Irene S

2012-03-01

Full Text Available Abstract Background The genome of Bacillus licheniformis DSM 13 harbours three neighbouring open reading frames showing protein sequence similarities to the proteins encoded from the Bacillus subtilis subsp. subtilis 168 gerA operon, GerAA, GerAB and GerAC. In B. subtilis, these proteins are assumed to form a germinant receptor involved in spore germination induced by the amino acid L-alanine. Results In this study we show that disruption of the gerAA gene in B. licheniformis MW3 hamper L-alanine and casein hydrolysate-triggered spore germination, measured by absorbance at 600 nm and confirmed by phase contrast microscopy. This ability was restored by complementation with a plasmid-borne copy of the gerA locus. Addition of D-alanine in the casein hydrolysate germination assay abolished germination of both B. licheniformis MW3 and the complementation mutant. Germination of both B. licheniformis MW3 and the gerA disruption mutant was induced by the non-nutrient germinant Ca2+-Dipicolinic acid. Conclusions These results demonstrate that the B. licheniformis MW3 gerA locus is involved in germination induced by L-alanine and potentially other components present in casein hydrolysate.

Role of the gerA operon in L-alanine germination of Bacillus licheniformis spores

Science.gov (United States)

2012-01-01

Background The genome of Bacillus licheniformis DSM 13 harbours three neighbouring open reading frames showing protein sequence similarities to the proteins encoded from the Bacillus subtilis subsp. subtilis 168 gerA operon, GerAA, GerAB and GerAC. In B. subtilis, these proteins are assumed to form a germinant receptor involved in spore germination induced by the amino acid L-alanine. Results In this study we show that disruption of the gerAA gene in B. licheniformis MW3 hamper L-alanine and casein hydrolysate-triggered spore germination, measured by absorbance at 600 nm and confirmed by phase contrast microscopy. This ability was restored by complementation with a plasmid-borne copy of the gerA locus. Addition of D-alanine in the casein hydrolysate germination assay abolished germination of both B. licheniformis MW3 and the complementation mutant. Germination of both B. licheniformis MW3 and the gerA disruption mutant was induced by the non-nutrient germinant Ca2+-Dipicolinic acid. Conclusions These results demonstrate that the B. licheniformis MW3 gerA locus is involved in germination induced by L-alanine and potentially other components present in casein hydrolysate. PMID:22420404

Production of D-alanine from DL-alanine using immobilized cells of Bacillus subtilis HLZ-68.

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Zhang, Yangyang; Li, Xiangping; Zhang, Caifei; Yu, Xiaodong; Huang, Fei; Huang, Shihai; Li, Lianwei; Liu, Shiyu

2017-09-13

Immobilized cells of Bacillus subtilis HLZ-68 were used to produce D-alanine from DL-alanine by asymmetric degradation. Different compounds such as polyvinyl alcohol and calcium alginate were employed for immobilizing the B. subtilis HLZ-68 cells, and the results showed that cells immobilized using a mixture of these two compounds presented higher L-alanine degradation activity, when compared with free cells. Subsequently, the effects of different concentrations of polyvinyl alcohol and calcium alginate on L-alanine consumption were examined. Maximum L-alanine degradation was exhibited by cells immobilized with 8% (w/v) polyvinyl alcohol and 2% (w/v) calcium alginate. Addition of 400 g of DL-alanine (200 g at the beginning of the reaction and 200 g after 30 h of incubation) into the reaction solution at 30 °C, pH 6.0, aeration of 1.0 vvm, and agitation of 400 rpm resulted in complete L-alanine degradation within 60 h, leaving 185 g of D-alanine in the reaction solution. The immobilized cells were applied for more than 15 cycles of degradation and a maximum utilization rate was achieved at the third cycle. D-alanine was easily extracted from the reaction solution using cation-exchange resin, and the chemical and optical purity of the extracted D-alanine was 99.1 and 99.6%, respectively.

Characterization of a novel variant of amino acid transport system asc in erythrocytes from Przewalski's horse (Equus przewalskii).

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Fincham, D A; Ellory, J C; Young, J D

1992-08-01

In thoroughbred horses, red blood cell amino acid transport activity is Na(+)-independent and controlled by three codominant genetic alleles (h, l, s), coding for high-affinity system asc1 (L-alanine apparent Km for influx at 37 degrees C congruent to 0.35 mM), low-affinity system asc2 (L-alanine Km congruent to 14 mM), and transport deficiency, respectively. The present study investigated amino acid transport mechanisms in red cells from four wild species: Przewalski's horse (Equus przewalskii), Hartmann's zebra (Zebra hartmannae), Grevy's zebra (Zebra grevyi), and onager (Equus hemonius). Red blood cell samples from different Przewalski's horses exhibited uniformly high rates of L-alanine uptake, mediated by a high-affinity asc1-type transport system. Mean apparent Km and Vmax values (+/- SE) for L-alanine influx at 37 degrees C in red cells from 10 individual animals were 0.373 +/- 0.068 mM and 2.27 +/- 0.11 mmol (L cells.h), respectively. As in thoroughbreds, the Przewalski's horse transporter interacted with dibasic as well as neutral amino acids. However, the Przewalski asc1 isoform transported L-lysine with a substantially (6.4-fold) higher apparent affinity than its thoroughbred counterpart (Km for influx 1.4 mM at 37 degrees C) and was also less prone to trans-stimulation effects. The novel high apparent affinity of the Przewalski's horse transporter for L-lysine provides additional key evidence of functional and possible structural similarities between asc and the classical Na(+)-dependent system ASC and between these systems and the Na(+)-independent dibasic amino acid transport system y+. Unlike Przewalski's horse, zebra red cells were polymorphic with respect to L-alanine transport activity, showing high-affinity or low-affinity saturable mechanisms of L-alanine uptake. Onager red cells transported this amino acid with intermediate affinity (apparent Km for influx 3.0 mM at 37 degrees C). Radiation inactivation analysis was used to estimate the target

A study of the irradiation temperature coefficient for L-alanine and DL-alanine dosemeters

International Nuclear Information System (INIS)

Desrosiers, M. F.; Lin, M.; Cooper, S. L.; Cui, Y.; Chen, K.

2006-01-01

Alanine dosimetry is now well established both as a reference and routine dosemeter for industrial irradiation processing. Accurate dosimetry under the relatively harsh conditions of industrial processing requires a characterisation of the parameters that influence the dosemeter response. The temperature of the dosemeter during irradiation is a difficult quantity to measure so that the accuracy of the temperature coefficient that governs the dosemeter response becomes a critical factor. Numerous publications have reported temperature coefficients for several types of alanine dosemeters. The observed differences in the measured values were commonly attributed to the differences in the polymer binder or the experimental design of the measurement. However, the data demonstrated a consistent difference in the temperature coefficients between L-alanine and DL-alanine. Since there were no commonalities in the dosemeter composition or the measurement methods applied, a clear conclusion is not possible. To resolve this issue, the two isomeric forms of alanine dosemeters were prepared and irradiated in an identical manner. The results indicated that the DL-alanine temperature coefficient is more than 50% higher than the L-alanine temperature coefficient. (authors)

Synthesis and characterization of new polyamides derived from alanine and valine derivatives

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El-Faham Ayman

2012-11-01

Full Text Available Abstract Background Many efforts have been recently devoted to design, investigate and synthesize biocompatible, biodegradable polymers for applications in medicine for either the fabrication of biodegradable devices or as drug delivery systems. Many of them consist of condensation of polymers having incorporated peptide linkages susceptible to enzymatic cleavage. Polyamides (PAs containing α-amino acid residues such as L-leucine, L-alanine and L-phenylalanine have been reported as biodegradable materials. Furthermore, polyamides (PAs derived from C10 and C14 dicarboxylic acids and amide-diamines derived from 1,6-hexanediamine or 1,12-dodecanediamine and L-phenylalanine, L-valyl-L-phenylalanine or L-phenylalanyl-L-valine residues have been reported as biocompatible polymers. We have previously described the synthesis and thermal properties of a new type of polyamides-containing amino acids based on eight new symmetric meta-oriented protected diamines derived from coupling of amino acids namely; Fomc-glycine, Fmoc-alanine, Fomc-valine and Fomc-leucine with m-phenylene diamine or 2,6-diaminopyridine. Results revealed that incorporation of pyridine onto the polymeric backbone of all series decreases the thermal stability. Here we describe another family of polyamides based on benzene dicarboxylic acid, pyridine dicarboxylic acid, and α-amino acid linked to benzidine and 4,4′-oxydianiline to study the effect of the dicarboxylic acid as well as the amino acids on the nature and thermal stability of the polymers. Results We report here the preparation of a new type of polyamides based on benzene dicarboxylic acid, pyridine dicarboxylic acid, and α-amino acid linked to benzidine and 4,4′-oxydianiline to study the effect of the dicarboxylic acid as well as the amino acids on the nature and thermal stability of polymers. The thermal properties of the polymers were evaluated by different techniques. Results revealed that structure-thermal property

and γγ-turns in proteins revisited: A new set of amino acid turn-type de

Indian Academy of Sciences (India)

mine, valine, glutamic acid and alanine has decreased for β-turns. Certain new amino acid preferences were observed for both turn types and individual amino acids showed turn-type dependent positional preferences. The rationale for new amino acid preferences are discussed in the light of hydrogen bonds and other.

Active Sites of Spinoxin, a Potassium Channel Scorpion Toxin, Elucidated by Systematic Alanine Scanning.

Science.gov (United States)

Peigneur, Steve; Yamaguchi, Yoko; Kawano, Chihiro; Nose, Takeru; Nirthanan, Selvanayagam; Gopalakrishnakone, Ponnampalam; Tytgat, Jan; Sato, Kazuki

2016-05-31

Peptide toxins from scorpion venoms constitute the largest group of toxins that target the voltage-gated potassium channel (Kv). Spinoxin (SPX) isolated from the venom of scorpion Heterometrus spinifer is a 34-residue peptide neurotoxin cross-linked by four disulfide bridges. SPX is a potent inhibitor of Kv1.3 potassium channels (IC50 = 63 nM), which are considered to be valid molecular targets in the diagnostics and therapy of various autoimmune disorders and cancers. Here we synthesized 25 analogues of SPX and analyzed the role of each amino acid in SPX using alanine scanning to study its structure-function relationships. All synthetic analogues showed similar disulfide bond pairings and secondary structures as native SPX. Alanine replacements at Lys(23), Asn(26), and Lys(30) resulted in loss of activity against Kv1.3 potassium channels, whereas replacements at Arg(7), Met(14), Lys(27), and Tyr(32) also largely reduced inhibitory activity. These results suggest that the side chains of these amino acids in SPX play an important role in its interaction with Kv1.3 channels. In particular, Lys(23) appears to be a key residue that underpins Kv1.3 channel inhibition. Of these seven amino acid residues, four are basic amino acids, suggesting that the positive electrostatic potential on the surface of SPX is likely required for high affinity interaction with Kv1.3 channels. This study provides insight into the structure-function relationships of SPX with implications for the rational design of new lead compounds targeting potassium channels with high potency.

Identification and elucidation of in vivo function of two alanine racemases from Pseudomonas putida KT2440.

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Duque, Estrella; Daddaoua, Abdelali; Cordero, Baldo F; De la Torre, Jesús; Antonia Molina-Henares, Maria; Ramos, Juan-Luis

2017-10-01

The genome of Pseudomonas putida KT2440 contains two open reading frames (ORFs), PP_3722 and PP_5269, that encode proteins with a Pyridoxal phosphate binding motif and a high similarity to alanine racemases. Alanine racemases play a key role in the biosynthesis of D-alanine, a crucial amino acid in the peptidoglycan layer. For these ORFs, we generated single and double mutants and found that inactivation of PP_5269 resulted in D-alanine auxotrophy, while inactivation of PP_3722 did not. Furthermore, as expected, the PP_3722/PP_5269 double mutant was a strict auxotroph for D-alanine. These results indicate that PP_5269 is an alr allele and that it is the essential alanine racemase in P. putida. We observed that the PP_5269 mutant grew very slowly, while the double PP_5269/PP_3722 mutant did not grow at all. This suggests that PP_3722 may replace PP_5269 in vivo. In fact, when the ORF encoding PP_3772 was cloned into a wide host range expression vector, ORF PP_3722 successfully complemented P. putida PP_5269 mutants. We purified both proteins to homogeneity and while they exhibit similar K M values, the V max of PP_5269 is fourfold higher than that of PP_3722. Here, we propose that PP_5269 and PP_3722 encode functional alanine racemases and that these genes be named alr-1 and alr-2 respectively. © 2017 Society for Applied Microbiology and John Wiley & Sons Ltd.

Polymers with complexing properties. Simple poly(amino acids)

Science.gov (United States)

Roque, J. M.

1978-01-01

The free amino (0.3 equiv/residue) and carboxyl (0.5 equiv/residue) groups of thermal polylysine increased dramatically on treatment with distilled water. The total hydrolysis of such a polymer was abnormal in that only about 50% of the expected amino acids were recovered. Poly (lysine-co-alanine-co-glycine) under usual conditions hydrolyzed completely in 8 hours; whereas, when it was pretreated with diazomethane, a normal period of 24 hours was required to give (nearly) the same amounts of each free amino acid as compared with those obtained from the untreated polymer. The amino groups of the basic thermal poly(amino acids) were sterically hindered. The existence of nitrogen atoms linking two or three chains and reactive groups (anhydride, imine) were proposed.

Invariant amino acids in the Mur peptide synthetases of bacterial peptidoglycan synthesis and their modification by site-directed mutagenesis in the UDP-MurNAc:L-alanine ligase from Escherichia coli.

Science.gov (United States)

Bouhss, A; Mengin-Lecreulx, D; Blanot, D; van Heijenoort, J; Parquet, C

1997-09-30

The comparison of the amino acid sequences of 20 cytoplasmic peptidoglycan synthetases (MurC, MurD, MurE, MurF, and Mpl) from various bacterial organisms has allowed us to detect common invariants: seven amino acids and the ATP-binding consensus sequence GXXGKT/S all at the same position in the alignment. The Mur synthetases thus appeared as a well-defined class of closely functionally related proteins. The conservation of a constant backbone length between certain invariants suggested common structural motifs. Among the other enzymes catalyzing a peptide bond formation driven by ATP hydrolysis to ADP and Pi, only folylpoly-gamma-l-glutamate synthetases presented the same common conserved amino acid residues, except for the most N-terminal invariant D50. Site-directed mutageneses were carried out to replace the K130, E174, H199, N293, N296, R327, and D351 residues by alanine in the MurC protein from Escherichia coli taken as model. For this purpose, plasmid pAM1005 was used as template, MurC being highly overproduced in this genetic setting. Analysis of the Vmax values of the mutated proteins suggested that residues K130, E174, and D351 are essential for the catalytic process whereas residues H199, N293, N296, and R327 were not. Mutations K130A, H199A, N293A, N296A, and R327A led to important variations of the Km values for one or more substrates, thereby indicating that these residues are involved in the structure of the active site and suggesting that the binding order of the substrates could be ATP, UDP-MurNAc, and alanine. The various mutated murC plasmids were tested for their effects on the growth, cell morphology, and peptidoglycan cell content of a murC thermosensitive strain at 42 degrees C. The observed effects (complementation, altered morphology, and reduced peptidoglycan content) paralleled more or less the decreased values of the MurC activity of each mutant.

Association between Serum Uric Acid and Elevated Alanine Aminotransferase in the General Population.

Science.gov (United States)

Chen, Shuang; Guo, Xiaofan; Yu, Shasha; Sun, Guozhe; Yang, Hongmei; Li, Zhao; Sun, Yingxian

2016-08-24

Both the serum uric acid (SUA) level and elevated alanine aminotransferase (ALT) are related to metabolic syndrome. However, the association between SUA and elevated ALT has not been elucidated in the general population. The objective of this study was to investigate the association between SUA and elevated ALT in the general population of China; A total of 11,572 adults (≥35 years of age) participated in this survey. Elevated ALT was defined as >40 U/L. SUA ≥ 7.0 mg/dL in males or ≥6.0 mg/dL in females was defined as hyperuricemia. SUA within the reference range was divided into quartiles, and its associations with elevated ALT were evaluated by logistic regressions; A total of 7.4% participants had elevated ALT. The prevalence of hyperuricemia was 14.9% in males and 7.3% in females. There was a significantly positive dose-response association between SUA levels and the prevalence of elevated ALT. After adjusting for potential confounders, a positive relationship for elevated ALT was observed in subjects with hyperuricemia (odds ratio [OR]: 2.032, 95% confidence interval [CI]: 1.443-2.861 for men; OR: 2.045, 95% CI: 1.221-3.425 for women, both p < 0.05). Within the reference range, the association between SUA and elevated ALT persisted in the fourth quartile (OR: 1.467, 95% CI: 1.063-2.025 for men; OR: 1.721, 95% CI: 1.146-2.585 for women, both p < 0.05); Our results indicated that an increased SUA level, even within the reference range, was independently associated with elevated ALT in Chinese adults.

Alanine racemase is essential for the growth and interspecies competitiveness of Streptococcus mutans.

Science.gov (United States)

Wei, Yuan; Qiu, Wei; Zhou, Xue-Dong; Zheng, Xin; Zhang, Ke-Ke; Wang, Shi-Da; Li, Yu-Qing; Cheng, Lei; Li, Ji-Yao; Xu, Xin; Li, Ming-Yun

2016-12-16

D-alanine (D-Ala) is an essential amino acid that has a key role in bacterial cell wall synthesis. Alanine racemase (Alr) is a unique enzyme that interconverts L-alanine and D-alanine in most bacteria, making this enzyme a potential target for antimicrobial drug development. Streptococcus mutans is a major causative factor of dental caries. The factors involved in the survival, virulence and interspecies interactions of S. mutans could be exploited as potential targets for caries control. The current study aimed to investigate the physiological role of Alr in S. mutans. We constructed alr mutant strain of S. mutans and evaluated its phenotypic traits and interspecies competitiveness compared with the wild-type strain. We found that alr deletion was lethal to S. mutans. A minimal supplement of D-Ala (150 μg·mL -1 ) was required for the optimal growth of the alr mutant. The depletion of D-alanine in the growth medium resulted in cell wall perforation and cell lysis in the alr mutant strain. We also determined the compromised competitiveness of the alr mutant strain relative to the wild-type S. mutans against other oral streptococci (S. sanguinis or S. gordonii), demonstrated using either conditioned medium assays or dual-species fluorescent in situ hybridization analysis. Given the importance and necessity of alr to the growth and competitiveness of S. mutans, Alr may represent a promising target to modulate the cariogenicity of oral biofilms and to benefit the management of dental caries.

Averaged electron collision cross sections for thermal mixtures of β-alanine conformers in the gas phase

Science.gov (United States)

Fujimoto, Milton M.; de Lima, Erik V. R.; Tennyson, Jonathan

2017-10-01

A theoretical study of elastic electron scattering by gas-phase amino acid molecule β-alanine (NH2-CH2-CH2-COOH) is presented. R-matrix calculations are performed for each of the ten lowest-lying, thermally-accessible conformers of β-alanine. Eigenphase sums, resonance features, differential and integral cross sections are computed for each conformer. The positions of the low-energy shape resonance associated with the unoccupied {π }* orbital of the -COOH group are found to vary from 2.5 to 3.3 eV and the resonance widths from 0.2 to 0.5 eV depending on the conformation. The temperature-dependent population ratios are derived, based on temperature-corrected Gibbs free energies. Averaged cross sections for thermal mixtures of the 10 conformers are presented. A comparison with previous results for the α-alanine isomer is also presented.

Thermodynamic Study of the Interaction of Bovine Serum Albumin and Amino Acids with Cellulose Nanocrystals.

Science.gov (United States)

Lombardo, Salvatore; Eyley, Samuel; Schütz, Christina; van Gorp, Hans; Rosenfeldt, Sabine; Van den Mooter, Guy; Thielemans, Wim

2017-06-06

The interaction of bovine serum albumin (BSA) with sulfated, carboxylated, and pyridinium-grafted cellulose nanocrystals (CNCs) was studied as a function of the degree of substitution by determining the adsorption isotherm and by directly measuring the thermodynamics of interaction. The adsorption of BSA onto positively charged pyridinium-grafted cellulose nanocrystals followed Langmuirian adsorption with the maximum amount of adsorbed protein increasing linearly with increasing degree of substitution. The binding mechanism between the positively charged pyridinum-grafted cellulose nanocrystals and BSA was found to be endothermic and based on charge neutralization. A positive entropy of adsorption associated with an increase of the degree of disorder upon addition of BSA compensated for the unfavorable endothermic enthalpy and enabled formation of pyridinium-g-CNC-BSA complexes. The endothermic enthalpy of adsorption was further found to decrease as a function of increasing degree of substitution. Negatively charged cellulose nanocrystals bearing sulfate and/or carboxylic functionalities were found to not interact significantly with the BSA protein. To investigate in more detail the role of single amino acids in the adsorption of proteins onto cellulose nanocrystals, we also studied the interaction of different types of amino acids with CNCs, i.e., charged (lysine, aspartic acid), aromatic (tryptophan, tyrosine), and polar (serine) amino acids. We found that none of the single amino acids bound with CNCs irrespective of surface charge and that therefore the binding of proteins with CNCs appears to require larger amino acid sequences that induce a greater entropic contribution to stabilize binding. Single amino acids are thus not adsorbed onto cellulose nanocrystals.

Controlled radical polymerization of an acrylamide containing L-alanine moiety via ATRP.

Science.gov (United States)

Rafiee, Zahra

2016-02-01

Homopolymerization of an optically active acrylamide having an amino acid moiety in the side chain, N-acryloyl-L-alanine (AAla) was carried out via atom transfer radical polymerization (ATRP) at room temperature using 2-hydroxyethyl-2'-methyl-2'-bromopropionate (HMB) or sodium-4-(bromomethyl)benzoate (SBB) as initiator in pure water, methanol/water mixture and pure methanol solvents. The polymerization reaction resulted in the optically active biocompatible amino acid-based homopolymer in good yield with narrow molecular weight distribution. The number average molecular weight increased with conversion and polydispersity was low. The structure and molecular weight of synthesized polymer were characterized by (1)H NMR, FT-IR spectroscopic techniques and size-exclusion chromatography.

Comparison of alanine dosimeters using silicone as their binder to a commercial, polystyrene-bound, alanine dosimeter

International Nuclear Information System (INIS)

Galindo, S.; Urena-Nunez, F.

1997-01-01

The feasibility of practical boron-containing alanine ESR dosimeters for gamma-neutron mixed field irradiation dosimeters depends in part on whether the γ response characteristics of these silicone-bound dosimeters are comparable to those of a commercially available dosimeter that has been used by the International Atomic Energy Agency (International Dose Assurance Service) as a transfer reference dosimeter. This work presents the results of the comparison of 3 batches of silicone-bound alanine dosimeters. The first batch consists of a mixture of alanine and boric acid; the second, alanine and borax; and the last contains only alanine. Results indicate that γ response characteristics of the silicone-bound samples are comparable to those of the commercial, polystyrene-bound, alanine dosimeter and that silicone has a strong potential as a binding substance for alanine ESR dosimetry. (Author)

Selection of D-Alanine-Tolerant Rice Cells

OpenAIRE

Hisashi, Manabe; Koji, Ohira; Aizu Junior College of Fukushima Prefecture; Department of Agricultural Chemistry, Faculty of Agriculture, Tohoku University

1984-01-01

By repeating subculture of rice cells (parent cells) in a D-alanine containing medium, we could select rice cells which grew well in the D-alanine medium. The D-alanine-tolerant cells absorbed a fairly small amount of D-alanine from the medium and did not accumulate much D-alanine in the cells. Aggregation of D-alanine-tolerant cells was greater than that of parent cells. D-Alanine metabolism of D-alanine.-tolerant cells did not increase in comparison with parent cells.

The Nitrogen Moieties of Dietary Nonessential Amino Acids Are Distinctively Metabolized in the Gut and Distributed to the Circulation in Rats.

Science.gov (United States)

Nakamura, Hidehiro; Kawamata, Yasuko; Kuwahara, Tomomi; Sakai, Ryosei

2017-08-01

Background: Although previous growth studies in rodents have indicated the importance of dietary nonessential amino acids (NEAAs) as nitrogen sources, individual NEAAs have different growth-promoting activities. This phenomenon might be attributable to differences in the nitrogen metabolism of individual NEAAs. Objective: The aim of this study was to compare nitrogen metabolism across dietary NEAAs with the use of their 15 N isotopologues. Methods: Male Fischer rats (8 wk old) were given 1.0 g amino acid-defined diets containing either 15 N-labeled glutamate, glutamine (amino or amide), aspartate, alanine, proline, glycine, or serine hourly for 5-6 h. Then, steady-state amino acid concentrations and their 15 N enrichments in the gut and in portal and arterial plasma were measured by an amino acid analyzer and LC tandem mass spectrometry, respectively. Results: The intestinal 15 N distribution and portal-arterial balance of 15 N metabolites indicated that most dietary glutamate nitrogen (>90% of dietary input) was incorporated into various amino acids, including alanine, proline, and citrulline, in the gut. Dietary aspartate nitrogen, alanine nitrogen, and amino nitrogen of glutamine were distributed similarly to other amino acids both in the gut and in the circulation. In contrast, incorporation of the nitrogen moieties of dietary proline, serine, and glycine into other amino acids was less than that of other NEAAs, although interconversion between serine and glycine was very active. Cluster analysis of 15 N enrichment data also indicated that dietary glutamate nitrogen, aspartate nitrogen, alanine nitrogen, and the amino nitrogen of glutamine were distributed similarly to intestinal and circulating amino acids. Further, the analysis revealed close relations between intestinal and arterial 15 N enrichment for each amino acid. The steady-state 15 N enrichment of arterial amino acids indicated that substantial amounts of circulating amino acid nitrogen are derived

Diverse taxa of cyanobacteria produce β-N-methylamino-l-alanine, a neurotoxic amino acid

OpenAIRE

Cox, Paul Alan; Banack, Sandra Anne; Murch, Susan J.; Rasmussen, Ulla; Tien, Georgia; Bidigare, Robert Richard; Metcalf, James S.; Morrison, Louise F.; Codd, Geoffrey A.; Bergman, Birgitta

2005-01-01

Cyanobacteria can generate molecules hazardous to human health, but production of the known cyanotoxins is taxonomically sporadic. For example, members of a few genera produce hepatotoxic microcystins, whereas production of hepatotoxic nodularins appears to be limited to a single genus. Production of known neurotoxins has also been considered phylogenetically unpredictable. We report here that a single neurotoxin, β-N-methylamino-l-alanine, may be produced by all known groups of cyanobacteria...

Comparison of EPR response of alanine and Gd₂O₃-alanine dosimeters exposed to TRIGA Mainz reactor.

Science.gov (United States)

Marrale, M; Schmitz, T; Gallo, S; Hampel, G; Longo, A; Panzeca, S; Tranchina, L

2015-12-01

In this work we report some preliminary results regarding the analysis of electron paramagnetic resonance (EPR) response of alanine pellets and alanine pellets added with gadolinium used for dosimetry at the TRIGA research reactor in Mainz, Germany. Two set-ups were evaluated: irradiation inside PMMA phantom and irradiation inside boric acid phantom. We observed that the presence of Gd2O3 inside alanine pellets increases the EPR signal by a factor of 3.45 and 1.24 in case of PMMA and boric acid phantoms, respectively. We can conclude that in the case of neutron beam with a predominant thermal neutron component the addition of gadolinium oxide can significantly improve neutron sensitivity of alanine pellets. Monte Carlo (MC) simulations of both response of alanine and Gd-added alanine pellets with FLUKA code were performed and a good agreement was achieved for pure alanine dosimeters. For Gd2O3-alanine deviations between MC simulations and experimental data were observed and discussed. Copyright © 2015 Elsevier Ltd. All rights reserved.

Effect of Exogenous _D-Alanine on _D-Alanyl-_D-alanine Content in Leaf Blades of Oryza australiensis Domin

OpenAIRE

Hisashi, Manabe; Aizu Junior College of Fukushima Prefecture

1986-01-01

In seedlings of Oryza australiensis Domin (W0008), most of the D-alanyl-D-alanine was distributed in the leaf blades. In excised leaf blades of W0008, exogenous D-alanine was incorporated into D-alanyl-D-alanine irrespective of the light condition as in Sasanishiki. With cultivation in D-alanine medium for several days, the D-alanyl-D-alanine content in W0008 leaf blades was found to increase, but no other D-alanine-containing dipeptides such as D-alanylglycine or D-alanyl-L-alanine were dete...

Synthesis and characterization of new nanocomposites films using alanine-Cu-functionalized graphene oxide as nanofiller and PVA as polymeric matrix for improving of their properties

Science.gov (United States)

Abdolmaleki, Amir; Mallakpour, Shadpour; Karshenas, Azam

2017-09-01

In the synthesis of polymer-graphene nanocomposites, for improving properties of nanocomposites, two factors dispersion and strong interfacial interactions between graphene and the polymer, are essential. In the present work, poly(vinyl alcohol) PVA/GO-Cu-alanine nanocomposite films were manufactured using concentrations 0, 1, 3 and 5 wt% of GO-Cu-alanine in water solution. For this purpose, L-alanine amino acid was located on the surface and edges of GO through copper(II) ion as a coordinating function. Then, flexible PVA/GO-Cu-alanine nanocomposite films were fabricated using GO-Cu-alanine as filler and PVA as matrix. Due to the existence of affective interaction between GO-Cu-alanine and PVA matrix, the acquired PVA/GO-Cu-alanine nanocomposites demonstrated great thermal and mechanical properties. Properties of manufactured materials were characterized by Fourier transform infrared, X-ray photoelectron spectroscopies (XPS), X-ray diffraction (XRD), Thermal gravimetric analysis, elemental analysis, field emission scanning electron microscopy, transmission electron microscopy and energy dispersive X-ray spectroscopy (EDX).

The influence of non-radiation induced ESR background signal from paraffin-alanine probes for dosimetry in the radiotherapy dose range

International Nuclear Information System (INIS)

Wieser, A.; Lettau, C.; Fill, U.; Regulla, D.F.

1993-01-01

The yield of radicals induced by ionizing radiation in the amino acid alanine and its quantification by ESR spectroscopy has proven excellent reproducibility. Those radicals trapped in the crystal lattice are prevented from recombination providing a thermally very stable system. This allows alanine to be applied as a transfer dosemeter. With paraffin-alanine probes ESR dosimetry can be performed with a standard deviation of ± 0.5% in the dose range from 20 Gy up to 100 kGy. At 1 Gy dose level the error increases to ± 6%. This dose level is three orders of magnitude higher than the calculated detection threshold for alanine with modern X-band ESR spectrometers. It was found that the poor standard deviation at the 1 Gy dose level, is not mainly produced by a bad signal-to-noise ratio but by a variable non-radiation induced ESR background signal from the alanine probes within a batch. In the present study the main sources of error for ESR dosimetry in the dose range below 20 Gy were analyzed. The influences of the production process, UV light and humidity upon the ESR background signal from paraffin-alanine probes were investigated. Measurements are shown indicating a second stable structure of the alanine radical at room temperature. (author)

Enrichment of Non-Terrestrial L-Proteinogenic Amino Acids by Aqueous Alteration on the Tagish Lake Meteorite Parent Body

Science.gov (United States)

Glavin, Daniel P.; Elsila, Jamie E.; Burton, Aaron S.; Callahan, Michael P.; Dworkin, Jason P.; Herd, Christopher D. K.

2012-01-01

The distribution and isotopic and enantiomeric compositions of amino acids found in three distinct fragments of the Tagish Lake C2-type carbonaceous chondrite were investigated via liquid chromatography fluorescence detection time-of-flight mass spectrometry and gas chromatography isotope ratio mass spectrometry. Large L-enantiomeric excesses (L(sub ee) approx. 43 to 59%) of the a-hydrogen aspartic and glutamic amino acids were measured in Tagish Lake, whereas alanine, another alpha-hydrogen protein amino acid, was found to be nearly racemic (D approx. L) using both techniques. Carbon isotope measurements of D- and L-aspartic acid and D- and L-alanine in Tagish Lake fall well outside of the terrestrial range and indicate that the measured aspartic acid enantioenrichment is indigenous to the meteorite. Alternate explanations for the Lexcesses of aspartic acid such as interference from other compounds present in the sample, analytical biases, or terrestrial amino acid contamination were investigated and rejected. These results can be explained by differences in the solid-solution phase behavior of aspartic acid, which can form conglomerate enantiopure solids during crystallization, and alanine, which can only form racemic crystals.

Free amino acids in spider hemolymph.

Science.gov (United States)

Tillinghast, Edward K; Townley, Mark A

2008-11-01

We examined the free amino acid composition of hemolymph from representatives of five spider families with an interest in knowing if the amino acid profile in the hemolymph of orb-web-building spiders reflects the high demands for small organic compounds in the sticky droplets of their webs. In nearly all analyses, on both orb and non-orb builders, glutamine was the most abundant free amino acid. Glycine, taurine, proline, histidine, and alanine also tended to be well-represented in orb and non-orb builders. While indications of taxon-specific differences in amino acid composition were observed, it was not apparent that two presumptive precursors (glutamine, taurine) of orb web sticky droplet compounds were uniquely enriched in araneids (orb builders). However, total amino acid concentrations were invariably highest in the araneids and especially so in overwintering juveniles, even as several of the essential amino acids declined during this winter diapause. Comparing the data from this study with those from earlier studies revealed a number of discrepancies. The possible origins of these differences are discussed.

AlaScan: A Graphical User Interface for Alanine Scanning Free-Energy Calculations.

Science.gov (United States)

Ramadoss, Vijayaraj; Dehez, François; Chipot, Christophe

2016-06-27

Computation of the free-energy changes that underlie molecular recognition and association has gained significant importance due to its considerable potential in drug discovery. The massive increase of computational power in recent years substantiates the application of more accurate theoretical methods for the calculation of binding free energies. The impact of such advances is the application of parent approaches, like computational alanine scanning, to investigate in silico the effect of amino-acid replacement in protein-ligand and protein-protein complexes, or probe the thermostability of individual proteins. Because human effort represents a significant cost that precludes the routine use of this form of free-energy calculations, minimizing manual intervention constitutes a stringent prerequisite for any such systematic computation. With this objective in mind, we propose a new plug-in, referred to as AlaScan, developed within the popular visualization program VMD to automate the major steps in alanine-scanning calculations, employing free-energy perturbation as implemented in the widely used molecular dynamics code NAMD. The AlaScan plug-in can be utilized upstream, to prepare input files for selected alanine mutations. It can also be utilized downstream to perform the analysis of different alanine-scanning calculations and to report the free-energy estimates in a user-friendly graphical user interface, allowing favorable mutations to be identified at a glance. The plug-in also assists the end-user in assessing the reliability of the calculation through rapid visual inspection.

Application of the alanine detector to gamma-ray, X-ray and fast neutron dosimetry

International Nuclear Information System (INIS)

Waligorski, M.P.R.; Hansen, J.W.; Byrski, E.

1987-01-01

A dosimeter based on alanine has been developed at the INP in Krakow and at Risoe National Laboratory. Due to its near tissue-equivalence and stability of signal, measured using ESR spectrometry at room temperature, this free-radical amino-acid dosimetric system is particularly suitable for measuring X-ray, gamma-ray and fast neutron doses in the range 10-10 5 Gy. The relative effectiveness (with respect to 60 Co γ-rays) of the alanine dosimeter to 250 kVp X-rays and to cyclotron-produced fast neutrons (mean neutron energy 5.6 MeV) is measured to be 0.76± 0.06 and 0.60±0.05, respectively. The suitability of the alanine dosimeter for intercomparison gamma-ray dosimetry is also shown. The estimated absolute difference between 60 Co dosimetry at Risoe National Laboratory and at the Centre of Oncology in Krakow is about 5%, somewhat more than the experimental uncertainty. These results are based on ESR measurements performed in Krakow on about 25% of the exposed detectors. 28 refs., 2 figs., 3 tabs. (author)

21 CFR 172.540 - DL-Alanine.

Science.gov (United States)

2010-04-01

... 21 Food and Drugs 3 2010-04-01 2009-04-01 true DL-Alanine. 172.540 Section 172.540 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR HUMAN... Agents and Related Substances § 172.540 DL-Alanine. DL-Alanine (a racemic mixture of D- and L-alanine...

Cellular and Physiological Effects of Dietary Supplementation with β-Hydroxy-β-Methylbutyrate (HMB) and β-Alanine in Late Middle-Aged Mice.

Science.gov (United States)

Vallejo, Julian; Spence, Madoka; Cheng, An-Lin; Brotto, Leticia; Edens, Neile K; Garvey, Sean M; Brotto, Marco

2016-01-01

There is growing evidence that severe decline of skeletal muscle mass and function with age may be mitigated by exercise and dietary supplementation with protein and amino acid ingredient technologies. The purposes of this study were to examine the effects of the leucine catabolite, beta-hydroxy-beta-methylbutyrate (HMB), in C2C12 myoblasts and myotubes, and to investigate the effects of dietary supplementation with HMB, the amino acid β-alanine and the combination thereof, on muscle contractility in a preclinical model of pre-sarcopenia. In C2C12 myotubes, HMB enhanced sarcoplasmic reticulum (SR) calcium release beyond vehicle control in the presence of all SR agonists tested (KCl, PHMB also improved C2C12 myoblast viability (25 μM HMB, P = 0.03) and increased proliferation (25 μM HMB, P = 0.04; 125 μM HMB, PHMB, diet with β-alanine, or diet with HMB and β-alanine. In β-alanine fed mice, EDL muscle showed a 7% increase in maximum absolute force compared to the control diet (202 ± 3vs. 188± 5 mN, P = 0.02). At submaximal frequency of stimulation (20 Hz), EDL from mice fed HMB plus β-alanine showed an 11% increase in absolute force (88.6 ± 2.2 vs. 79.8 ± 2.4 mN, P = 0.025) and a 13% increase in specific force (12.2 ± 0.4 vs. 10.8 ± 0.4 N/cm2, P = 0.021). Also in EDL muscle, β-alanine increased the rate of force development at all frequencies tested (PHMB reduced the time to reach peak contractile force (TTP), with a significant effect at 80 Hz (P = 0.0156). In soleus muscle, all experimental diets were associated with a decrease in TTP, compared to control diet. Our findings highlight beneficial effects of HMB and β-alanine supplementation on skeletal muscle function in aging mice.

Manageable cytotoxicity of nanocapsules immobilizing D-amino acid oxidase via exogenous administration of nontoxic prodrug

Science.gov (United States)

Zhao, Yang; Zhu, Yingchun; Fu, Jingke

2014-02-01

D-Amino acid oxidase (DAO), which could catalyze generation of hydrogen peroxide with strong oxidbility and cytotoxicity, has become of interest as a biocatalyst for therapeutic treatments. Herein we report that amino-functional hollow mesoporous silica with large pore size (10.27 nm) and positively charged surface effectively immobilize DAO with negative charge. The adsorption, activity and stability of DAO are demonstrated to depend mainly on the amino-functionalization of surface. Significant cancer cell killing effect is observed when the cells are treated by the nanocapsules entrapping DAO together with D-alanine, showing distinct dose-dependency on concentration of the nanocapsules entrapping DAO or D-alanine. Nevertheless, the toxicity is completely neutralized by the addition of catalase, and anti-tumor effect is not observed when either the nanocapsules entrapping DAO or D-alanine is applied alone. The results indicate that cytotoxicity of the nanocapsules entrapping DAO could be managed by exogenous administration of nontoxic prodrug to tumor tissue, due to the stereoselectivity of DAO and the scarcity of its substrates in mammalian organisms. Thus, the method might be exploited as a potential treatment for cancer therapy.

21 CFR 582.5118 - Alanine.

Science.gov (United States)

2010-04-01

... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Alanine. 582.5118 Section 582.5118 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL DRUGS, FEEDS... § 582.5118 Alanine. (a) Product. Alanine (L- and DL-forms). (b) Conditions of use. This substance is...

A conserved residue of l-alanine dehydrogenase from Bacillus pseudofirmus, Lys-73, participates in the catalytic reaction through hydrogen bonding.

Science.gov (United States)

He, Guangzheng; Xu, Shujing; Wang, Shanshan; Zhang, Qing; Liu, Dong; Chen, Yuling; Ju, Jiansong; Zhao, Baohua

2018-03-01

A multiple protein sequence alignment of l-alanine dehydrogenases from different bacterial species revealed that five highly conserved amino acid residues Arg-15, Lys-73, Lys-75, His-96 and Asp-269 are potential catalytic residues of l-alanine dehydrogenase from Bacillus pseudofirmus OF4. In this study, recombinant OF4Ald and its mutants of five conserved residues were constructed, expressed in Escherichia coli, purified by His 6 -tag affinity column and gel filtration chromatography, structure homology modeling, and characterized. The purified protein OF4Ald displayed high specificity to l-alanine (15Umg -1 ) with an optimal temperature and pH of 40°C and 10.5, respectively. Enzymatic assay and activity staining in native gels showed that mutations at four conserved residue Arg-15, Lys-75, His-96 and Asp-269 (except residue Lys-73) resulted in a complete loss in enzymatic activity, which signified that these predicted active sites are indispensable for OF4Ald activity. In contrast, the mutant K73A resulted in 6-fold improvement in k cat /K m towards l-alanine as compared to the wild type protein. Further research of the residue Lys-73 substituted by various amino acids and structural modeling revealed that residue Lys-73 might be involved in the catalytic reaction of the enzyme by influencing the enzyme-substrate binding through the hydrogen-bonding interaction with conserved residue Lys-75. Copyright © 2017 Elsevier Inc. All rights reserved.

Study of EPR spectra of radicals from ionizing radiation interaction with alanine and 4-hydroxyproline samples

International Nuclear Information System (INIS)

Simion, Corina Anca; Georgescu, Rodica; Grigorescu, Eric Leon

2006-01-01

A number of stable chemical radicals result following irradiation with ionizing rays of α-β-alanine and 4-hydroxyproline. They could be put into evidence using post-irradiation EPR technique. Analysis and inter-comparison of spectra signals become important for a correct assignment of structure and, subsequently of generating mechanisms in amino acids irradiated samples. (author)

X-ray diffraction study on the structure of concentrated aqueous solutions involving alanine molecules with different optical activities

International Nuclear Information System (INIS)

Kameda, Yasuo; Okuyama, Aya; Amo, Yuko; Usuki, Takeshi; Kohara, Shinji

2007-01-01

X-ray diffraction measurements on aqueous 2.5 mol% DL-, L-, and D-alanine solutions in D 2 O were carried out at 26±2degC in order to obtain information concerning the difference in the hydrogen-bonded structure between aqueous solutions involving amino acid molecules with different optical activities. The difference function, Δi inter (Q), between intermolecular interference term observed for DL- and L-alanine and between DL- and D-alanine solutions both exhibited a first peak at Q=1.6 A -1 , followed by oscillatory features extending to higher-Q region, implying that there is a difference in the intermolecular structure is present between these solutions. The difference distribution function, Δg inter (r), obtained from the Fourier transform of the Δi inter (Q) between DL- and L-, and between DL- and D-alanine solutions showed an obvious negative peak at r=2.8 A, which was attributed to the nearest neighbor hydrogen-bonded O...O interaction. The least squares fitting analysis of the observed Δi inter (Q) showed that the intermolecular O...O distance and the difference in the coordination number between DL- and L-, and between DL- and D-alanine solutions are 2.76(2) A and -0.18(1), and 2.81(3) A and -0.18(1), respectively. It was concluded that the intermolecular hydrogen-bonded network in aqueous L- and D-alanine solutions is stronger than that in the DL-alanine solution. (author)

Solubility of the Proteinogenic α-Amino Acids in Water, Ethanol, and Ethanol-Water Mixtures

NARCIS (Netherlands)

Bowden, Nathan A.; Sanders, Johan P.M.; Bruins, Marieke E.

2018-01-01

The addition of organic solvents to α-amino acids in aqueous solution could be an effective method in crystallization. We reviewed the available data on the solubility of α-amino acids in water, water-ethanol mixtures, and ethanol at 298.15 K and 0.1 MPa. The solubility of l-alanine, l-proline,

Use of the entire spectrum of irradiated alanine for dosimetry

International Nuclear Information System (INIS)

Dolo, J.M.; Moignau, F.

2005-01-01

Alanine is an amino acid commonly used in ESR dosimetry as a reference detector. The classic approach for the measurement of irradiated samples is to determine the amplitude of the central peak of the first derivative spectrum. It is generally considered that this technique represents the best and most reproducible solution for achieving an accurate proportionality between the concentration of free radicals inside the resonant cavity, characterized by the amplitude, and the dose. It is also accepted that this central peak corresponds to the free radical CH 3 .CHCOO - . The hyperfine structure of this radical in the spectrum shows five main peaks with the approximate ratios 1:4:6:4:1 as regards coupling. This paper presents another approach featuring analysis of the entire spectrum: (i) ratios of identified peaks (ii) ratio variation vs time with regard to several parameters affecting fading. These variations in the alanine spectrum are probably correlated with the variation of the concentrations of different free radical species. These variations and their positions in the spectrum are very important constraints that increase the uncertainty of this type of measurement

Ultraviolet radiation induces stress in etiolated Landoltia punctata, as evidenced by the presence of alanine, a universal stress signal: a ¹⁵N NMR study.

Science.gov (United States)

Monselise, E B-I; Levkovitz, A; Kost, D

2015-01-01

Analysis with (15) N NMR revealed that alanine, a universal cellular stress signal, accumulates in etiolated duckweed plants exposed to 15-min pulsed UV light, but not in the absence of UV irradiation. The addition of 10 mm vitamin C, a radical scavenger, reduced alanine levels to zero, indicating the involvement of free radicals. Free D-alanine was detected in (15) N NMR analysis of the chiral amino acid content, using D-tartaric acid as solvent. The accumulation of D-alanine under stress conditions presents a new perspective on the biochemical processes taking place in prokaryote and eukaryote cells. © 2014 German Botanical Society and The Royal Botanical Society of the Netherlands.

Functional Characterization of Alanine Racemase from Schizosaccharomyces pombe: a Eucaryotic Counterpart to Bacterial Alanine Racemase

OpenAIRE

Uo, Takuma; Yoshimura, Tohru; Tanaka, Naotaka; Takegawa, Kaoru; Esaki, Nobuyoshi

2001-01-01

Schizosaccharomyces pombe has an open reading frame, which we named alr1+, encoding a putative protein similar to bacterial alanine racemase. We cloned the alr1+ gene in Escherichia coli and purified the gene product (Alr1p), with an Mr of 41,590, to homogeneity. Alr1p contains pyridoxal 5′-phosphate as a coenzyme and catalyzes the racemization of alanine with apparent Km and Vmax values as follows: for l-alanine, 5.0 mM and 670 μmol/min/mg, respectively, and for d-alanine, 2.4 mM and 350 μmo...

Solubility of the Proteinogenic α-Amino Acids in Water, Ethanol, and Ethanol–Water Mixtures

Science.gov (United States)

2018-01-01

The addition of organic solvents to α-amino acids in aqueous solution could be an effective method in crystallization. We reviewed the available data on the solubility of α-amino acids in water, water–ethanol mixtures, and ethanol at 298.15 K and 0.1 MPa. The solubility of l-alanine, l-proline, l-arginine, l-cysteine, and l-lysine in water and ethanol mixtures and the solubility of l-alanine, l-proline, l-arginine, l-cysteine, l-lysine, l-asparagine, l-glutamine, l-histidine, and l-leucine in pure ethanol systems were measured and are published here for the first time. The impact on the solubility of amino acids that can convert in solution, l-glutamic acid and l-cysteine, was studied. At lower concentrations, only the ninhydrin method and the ultraperfomance liquid chromatography (UPLC) method yield reliable results. In the case of α-amino acids that convert in solution, only the UPLC method was able to discern between the different α-amino acids and yields reliable results. Our results demonstrate that α-amino acids with similar physical structures have similar changes in solubility in mixed water/ethanol mixtures. The solubility of l-tryptophan increased at moderate ethanol concentrations. PMID:29545650

Studies on radiolysis of amino acids, (4)

International Nuclear Information System (INIS)

Oku, Tadatake

1978-01-01

In order to elucidate the effect of adding methionine on the loss of amino acid by γ-irradiation in amino acid mixture, because methionine is one of the most radio-sensitive in amino acids, the remaining amino acids in γ-irradiated aqueous solution of amino acid mixture were studied by determining the total amount of each remaining amino acid. The mixture of 18 amino acids which contains methionine and that of 17 amino acids without methionine were used. Amino acids and the irradiation products were determined with an automatic amino acid analyzer. The total amount of remaining amino acids in the irradiated solution of 18 amino acid mixture was more than that of 17 amino acid mixture. The order of the total amount of each remaining amino acid by low-dose irradiation was Gly>Ala>Asp>Glu>Val>Ser, Pro>Ile, Leu>Thr>Lys>Tyr>Arg>His>Phe>Try>Cys>Met. In case of the comparison of amino acids of same kinds, the total remaining amount of each amino acid in amino acid mixture was more than that of individually irradiated amino acid. The total remaining amounts of glycine, alanine and aspartic acid in irradiated 17 amino acid mixture resulted in slight increase. Ninhydrin positive products formed from 18 amino acid mixture irradiated with 2.640 x 10 3 rad were ammonia, methionine sulfoxide and DOPA of 1.34, 0.001 and 0.25 μmoles/ml of the irradiated solution, respectively. (Kobake, H.)

Comparative Potential Protect Effect of HSCAS, Diatomite and ...

African Journals Online (AJOL)

mdenli

Aspartate amino transferase (AST), alanine amino transferase (ALT) activities and total protein concentration in the serum ... consuming AFB1 at 80 µg/kg feed showed a yellowish colour, portal leucocytic infiltration, congestion, .... Data were analysed using the General Linear models (GLM) procedure of SPSS 9.0 (1993).

Effect of Calitropis Procera Aqueous Root Extract Against CCL4 ...

African Journals Online (AJOL)

ABUBAKAR

ABSTRACT. The hepatocurative effect of aqueous root extract of Calitropis Procera on CCl4 induced hepatotoxicity in rabbits was studied in groups of rabbit and the levels of liver enzymes; aspartate amino transferase (AST), alanine amino transferase (ALT) and alkaline phosphatase (ALP). Serum concentrations of total ...

Serum neuron-specific enolase, biogenic amino-acids and neurobehavioral function in lead-exposed workers from lead-acid battery manufacturing process.

Science.gov (United States)

Ravibabu, K; Barman, T; Rajmohan, H R

2015-01-01

The interaction between serum neuron-specific enolase (NSE), biogenic amino-acids and neurobehavioral function with blood lead levels in workers exposed to lead form lead-acid battery manufacturing process was not studied. To evaluate serum NSE and biogenic amino-acids (dopamine and serotonin) levels, and neurobehavioral performance among workers exposed to lead from lead-acid storage battery plant, and its relation with blood lead levels (BLLs). In a cross-sectional study, we performed biochemical and neurobehavioral function tests on 146 workers exposed to lead from lead-acid battery manufacturing process. BLLs were assessed by an atomic absorption spectrophotometer. Serum NSE, dopamine and serotonin were measured by ELISA. Neurobehavioral functions were assessed by CDC-recommended tests---simple reaction time (SRT), symbol digit substitution test (SDST), and serial digit learning test (SDLT). There was a significant correlation (r 0.199, pSDLT and SRT had also a significant positive correlation (r 0.238, p<0.01). NSE had a negative correlation (r -0.194, p<0.05) with serotonin level. Multiple linear regression analysis revealed that both SRT and SDST had positive significant associations with BLL. SRT also had a positive significant association with age. Serum NSE cannot be used as a marker for BLL. The only domain of neurobehavioral function tests that is affected by increased BLL in workers of lead-acid battery manufacturing process is that of the "attention and perception" (SDST).

Protein Nanopore-Based Discrimination between Selected Neutral Amino Acids from Polypeptides.

Science.gov (United States)

Asandei, Alina; Rossini, Aldo E; Chinappi, Mauro; Park, Yoonkyung; Luchian, Tudor

2017-12-19

Nanopore probing of biological polymers has the potential to achieve single-molecule sequencing at low cost, high throughput, portability, and minimal sample preparation and apparatus. In this article, we explore the possibility of discrimination between neutral amino acid residues from the primary structure of 30 amino acids long, engineered peptides, through the analysis of single-molecule ionic current fluctuations accompanying their slowed-down translocation across the wild type α-hemolysin (α-HL) nanopore, and molecular dynamics simulations. We found that the transient presence inside the α-HL of alanine or tryptophan residues from the primary sequence of engineered peptides results in distinct features of the ionic current fluctuation pattern associated with the peptide reversibly blocking the nanopore. We propose that α-HL sensitivity to the molecular exclusion at the most constricted region mediates ionic current blockade events correlated with the volumes that are occluded by at least three alanine or tryptophan residues, and provides the specificity needed to discriminate between groups of neutral amino acids. Further, we find that the pattern of current fluctuations depends on the orientation of the threaded amino acid residues, suggestive of a conformational anisotropy of the ensemble of conformations of the peptide on the restricted nanopore region, related to its relative axial orientation inside the nanopore.

Effects of Artemisia dracunculus Aqueous Extract on Blood Sugar, Serum Insulin, Triglyceride and Liver Enzymes in Fructose Drinking Water Male Rats

Directory of Open Access Journals (Sweden)

Mohammad Reza Shahraki

2017-02-01

Full Text Available Background Artemisia are various groups of plants which are used as an herbal medicine in all countries; the present study was designed to evaluate the effects of Artemisia dracunculus (AD leaves aqueous extract on blood sugar, serum insulin, and triglyceride and liver enzymes in Fructose Drinking water (FDW male rats. Methods At the beginning of experiment, 48 Wistar-albino male rats, weighing 200 - 250g were divided into control (C and FDW groups (n = 24. FDW group received FDW (10%, w/v for a month but control group did not receive any agents during the trial period. A half of control and FDW groups received AD L aqueous extract daily during trial period. At the end, animals were anesthetized, sacrificed and blood samples were collected from cervical vessels. Serum insulin, Blood glucose, insulin resistance index, triglyceride and liver enzymes were measured by ordinary methods. Obtained data were analyzed using SPSS-17 via one way ANOVA and Tukey tests. Results Our results showed that serum insulin, blood sugar, insulin resistance index, triglyceride, Aspartate amino transferase (AST and Alanine amino transferase (ALT values in FDW group significantly increased compared to C and C + E groups but these values in group FDW + E were significantly decreases compared to group FDW (P < 0.001. Conclusions Our findings demonstrated that AD L aqueous extract improves blood sugar, serum insulin, insulin resistance index and liver enzymes in rat model.

Highly Efficient Fumed Silica Nanoparticles for Peptide Bond Formation: Converting Alanine to Alanine Anhydride.

Science.gov (United States)

Guo, Chengchen; Jordan, Jacob S; Yarger, Jeffery L; Holland, Gregory P

2017-05-24

In this work, thermal condensation of alanine adsorbed on fumed silica nanoparticles is investigated using thermal analysis and multiple spectroscopic techniques, including infrared (IR), Raman, and nuclear magnetic resonance (NMR) spectroscopies. Thermal analysis shows that adsorbed alanine can undergo thermal condensation, forming peptide bonds within a short time period and at a lower temperature (∼170 °C) on fumed silica nanoparticle surfaces than that in bulk (∼210 °C). Spectroscopic results further show that alanine is converted to alanine anhydride with a yield of 98.8% during thermal condensation. After comparing peptide formation on solution-derived colloidal silica nanoparticles, it is found that fumed silica nanoparticles show much better efficiency and selectivity than solution-derived colloidal silica nanoparticles for synthesizing alanine anhydride. Furthermore, Raman spectroscopy provides evidence that the high efficiency for fumed silica nanoparticles is likely related to their unique surface features: the intrinsic high population of strained ring structures present at the surface. This work indicates the great potential of fumed silica nanoparticles in synthesizing peptides with high efficiency and selectivity.

Theoretical and experimental radiation effectiveness of the free radical dosimeter alanine to irradiation with heavy charged particles

DEFF Research Database (Denmark)

Hansen, Jørgen-Walther; Olsen, K. J.

1985-01-01

Dose-response characteristics have been measured for the crystalline amino acid L-.alpha.-alanine irradiated with ion beams of 6 and 16 MeV protons, 20 MeV .alpha. particles, 21 MeV7Li ions, 64 MeV16O ions, and 80 MeV32S ions. The experimental radiation effectiveness (RE) with reference to low-LE...

Serum metabolomics of Indian women with polycystic ovary syndrome using 1H NMR coupled with a pattern recognition approach.

Science.gov (United States)

RoyChoudhury, Sourav; Mishra, Biswa Prasanna; Khan, Tila; Chattopadhayay, Ratna; Lodh, Indrani; Datta Ray, Chaitali; Bose, Gunja; Sarkar, Himadri S; Srivastava, Sudha; Joshi, Mamata V; Chakravarty, Baidyanath; Chaudhury, Koel

2016-10-18

Polycystic ovary syndrome (PCOS) is one of the most commonly occurring metabolic and endocrinological disorders affecting women of reproductive age. Metabolomics is an emerging field that holds promise in understanding disease pathophysiology. Recently, a few metabolomics based studies have been attempted in PCOS patients; however, none of them have included patients from the Indian population. The main objective of this study was to investigate the serum metabolomic profile of Indian women with PCOS and compare them with controls. Proton nuclear magnetic resonance ( 1 H NMR) was used to first identify the differentially expressed metabolites among women with PCOS from the Eastern region of India during the discovery phase and further validated in a separate cohort of PCOS and control subjects. Multivariate analysis of the binned spectra indicated 16 dysregulated bins in the sera of these women with PCOS. Out of these 16 bins, 13 identified bins corresponded to 12 metabolites including 8 amino acids and 4 energy metabolites. Amongst the amino acids, alanine, valine, leucine and threonine and amongst the energy metabolites, lactate and acetate were observed to be significantly up-regulated in women with PCOS when compared with controls. The remaining 4 amino acids, l-glutamine, proline, glutamate and histidine were down-regulated along with 2 energy metabolites: glucose and 3-hydroxybutyric acid. Our findings showed dysregulations in the expression of different metabolites in the serum of women with PCOS suggesting the involvement of multiple pathways including amino acid metabolism, carbohydrate/lipid metabolism, purine and pyrimidine metabolism and protein synthesis.

Interaction of metal ions and amino acids - Possible mechanisms for the adsorption of amino acids on homoionic smectite clays

Science.gov (United States)

Gupta, A.; Loew, G. H.; Lawless, J.

1983-01-01

A semiempirical molecular orbital method is used to characterize the binding of amino acids to hexahydrated Cu(2+) and Ni(2+), a process presumed to occur when they are adsorbed in the interlamellar space of homoionic smectite clays. Five alpha-amino acids, beta-alanine, and gamma-aminobutyric acid were used to investigate the metal ion and amino acid specificity in binding. It was assumed that the alpha, beta, and gamma-amino acids would bind as bidentate anionic ligands, forming either 1:1 or 1:2 six-coordinated five, six, and seven-membered-ring chelate complexes, respectively. Energies of complex formation, optimized geometries, and electron and spin distribution were determined; and steric constraints of binding of the amino acids to the ion-exchanged cations in the interlamellar spacing of a clay were examined. Results indicate that hexahydrated Cu(2+) forms more stable complexes than hexahydrated Ni(2+) with all the amino acids studied. However, among these amino acids, complex formation does not favor the adsorption of the biological subset. Calculated energetics of complex formation and steric constraints are shown to predict that 1:1 rather than 1:2 metal-amino acid complexes are generally favored in the clay.

Procedure for preparation of 3-fluor-D-alanine, 2-deutero-3-fluor-D-alanine and 2,3,3-trideutero-3-fluor-D-alanine and their salts

International Nuclear Information System (INIS)

Kollonitsch, J.; Kahan, F.M.

1971-01-01

Procedures for the preparation of 3-fluor-D-alanine, 2-deutero-3-fluor-D-alanine and 2,3,3-trideutero-3-fluor-D-alanine, and salts of these compounds, are described. These new compounds are useful antibacterial substances not only applicable in the disinfection of pharmaceutical, dental and medical equipment, but also in the treatment of diseases caused by bacteria, and may be administered orally. While 3-fluor-L-alanine metabolises rapidly with toxic results, 3-fluor-D-alanine is much more slowly broken down in vivo and is not harmful in normal doses. Further it has been found that deuteration gives new deutero-analogues which are less subject to metabolic breaking down and still retain the antibacterial strength of the original compound. The in vivo activity is thereby increased and maintained. (JIW)

Photochemical exchange reactions of thymine, uracil and their nucleosides with selected amino acids

International Nuclear Information System (INIS)

Shetlar, M.D.; Taylor, J.A.; Hom, K.

1984-01-01

The photoinduced exchange reactions of thymine with lysine at basic pH, using 254 nm light, have been studied. Three products have been isolated, namely, 6-amino-2-(1-thyminyl)hexanoic acid (Ia), 2-amino-6-(1-thyminyl)hexanoic acid (IIa) and 1-amino-5-(1-thyminyl)pentane (IIIa). Compound IIIa was shown to be a secondary product, produced by photochemical decarboxylation of Ia. Photochemical reaction of thymine with glycine and alanine at basic pH led, respectively, to formation of 2-(1-thyminyl)acetic acid (Ic) and 2-(1-thyminyl)propionic acid (Id). Compounds Ic and Id underwent photolysis to produce the decarboxylated secondary products 1-methylthymine and 1-ethylthymine, respectively. Thymidine reacts photochemically with glycine and alanine to produce the same products. Irradiation of DNA in the presence of lysine at basic pH led to the formation of the same products formed in the thymine-lysine system, namely Ia, IIa and IIIa. Uracil was found to undergo analogous photochemical exchange reactions with lysine to form 6-amino-2-(1-uracilyl)hexanoic acid (Ib), and 2-amino-6-(1-uracilyl)hexanoic acid (IIb). Compound Ib was found to undergo photodecarboxylation to form 1-amino-5-(1-uracilyl)pentane (IIIb), analogous to the secondary photoreaction of Ia. Photoreaction of uracil with 1,5-diaminopentane (cadaverine) likewise led to formation of IIIb. (author)

Prognostic value of pretreatment serum alanine aminotransferase/aspartate aminotransferase (ALT/AST) ratio and gamma glutamyltransferase (GGT) in patients with esophageal squamous cell carcinoma.

Science.gov (United States)

Huang, Hao; Wang, Xue-Ping; Li, Xiao-Hui; Chen, Hao; Zheng, Xin; Lin, Jian-Hua; Kang, Ting; Zhang, Lin; Chen, Pei-Song

2017-08-14

The levels of liver function tests (LFTs) are often used to assess liver injury and non-liver disease-related mortality. In our study, the relationship between pretreatment serum LFTs and overall survival (OS) was evaluated in esophageal squamous cell carcinoma (ESCC) patients. Our purpose was to investigate the prognostic value of the preoperative alanine aminotransferase/aspartate aminotransferase (ALT/AST) ratio and gamma glutamyltransferase (GGT) in ESCC patients. A retrospective study was performed in 447 patients with ESCC, and follow-up period was at least 60 months until death. The prognostic significance of serum LFTs were determined by univariate and multivariate Cox hazard models. LFTs including ALT, AST, LSR, GGT, TBA and LDH were analyzed. Serum LSR (HR: 0.592, 95% CI = 0.457-0.768, p < 0.001 and GGT (HR: 1.507, 95% CI = 1.163-1.953, p = 0.002) levels were indicated as significant predictors of OS. The 5-year OS among patients with higher LSR levels was longer compared with those patients with decreased LSR levels, not only in the whole cohort but also in the subgroups stratified by pathological stage (T1-T2 subgroup, T3-T4 subgroup, N0 subgroup and M0 subgroup). We also found that patients with a higher GGT might predict worse OS than patients with a normal GGT, not only in the whole cohort but also in the subgroups stratified by pathological stage (T3-T4 subgroup and N1-N2 subgroup). Both increased levels of LSR and decreased levels of GGT might predict shorter overall survival in ESCC patients. Our findings suggest that serum LSR and GGT levels could be used as a key predictor of survival in patients with ESCC.

Growth, optical, thermal and dielectric studies of an amino acid organic nonlinear optical material: L-Alanine

International Nuclear Information System (INIS)

Caroline, M. Lydia; Sankar, R.; Indirani, R.M.; Vasudevan, S.

2009-01-01

Good transparent bulk single crystals of L-alanine (nonlinear optical material) have been grown successfully by slow cooling technique from aqueous solution at pH value of 2.0. Optically transparent crystals with dimensions 2.4 cm x 1.2 cm x 1.6 cm, were grown by optimizing the growth parameters within a growth period of 2 weeks. The crystallinity of L-alanine crystal was confirmed by the powder X-ray diffraction study and diffraction peaks are indexed. The vibrational structure of the molecule is elucidated from FTIR spectra. The thermal behaviour of the grown crystal was investigated by thermogravimetric (TG) and differential thermal analyses (DTA) techniques in a nitrogen atmosphere. The result showed that the material starts decomposing at 297 deg. C. Its optical behaviour has been examined by UV-vis spectral analysis, which shows the absence of absorbance between the wavelengths ranging from 200 to 1200 nm. The NLO property was confirmed by the powder technique of Kurtz and Perry. The dielectric behaviour of the sample was also studied for the first time

Alanine flux in obese and healthy humans as evaluated by 15N- and 2H3-labeled alanines

International Nuclear Information System (INIS)

Hoffer, L.J.; Yang, R.D.; Matthews, D.E.; Bistrian, B.R.; Bier, D.M.; Young, V.R.

1988-01-01

Estimates of plasma alanine flux as measured in humans using L-[ 15 N]-alanine or L-[3,3,3- 2 H 3 ]alanine were compared by simultaneous intravenous infusion of both tracers. Plasma isotope enrichments were measured by chemical ionization gas chromatography-mass spectrometry. In 16 obese women before and during a hypocaloric diet and in 4 normal men in the postabsorptive and fed states, the fluxes were highly correlated (r2 = 0.93) although plasma alanine flux with the 2 H tracer was two to three times greater than that obtained with [ 15 N]alanine. The fluxes decreased with the hypocaloric diet in obese subjects and increased during the fed state in healthy adults. Thus, although the estimates of alanine flux differed according to the tracer used, both appear to give equivalent information about changes in alanine kinetics induced by the nutritional conditions examined

Urinary and Serum Metabolomics Analyses Uncover That Total Glucosides of Paeony Protect Liver against Acute Injury Potentially via Reprogramming of Multiple Metabolic Pathways

Directory of Open Access Journals (Sweden)

Haojie Li

2017-01-01

Full Text Available Total glucosides of paeony (TGP have been confirmed to be hepatoprotective. However, the underlying mechanism is largely unclear. In this study, we investigated the metabolic profiles of urine and serum in rats with carbon tetrachloride- (CCl4- induced experimental liver injury and TGP administration by using ultra-performance liquid chromatography-mass spectrometry (UPLC-MS. The vehicle or a single dose of TGP was intragastrically administered to Wistar rats once a day for 14 consecutive days. To induce ALI, 50% CCl4 was injected intraperitoneally into these rats 2 hours after the last time administration of saline of TGP at the 14th day. The results indicated that TGP administration could protect rats from CCl4-induced ALI and alanine aminotransferase (ALT and aspartate aminotransferase (AST elevation, as well as hepatocyte apoptosis and inflammation. Furthermore, metabolomics analysis showed that TGP treatment significantly attenuated CCl4-triggered deregulation of multiple metabolites in both urine and serum, including glycine, alanine, proline, and glutamine. Metabolite set enrichment and pathway analyses demonstrated that amino acid cycling and glutathione metabolism were two main pathways involved in CCl4-induced experimental liver injury and TGP administration. Taken together, these findings revealed that regulation of metabolites potentially plays a pivotal role in the protective effect of TGP on ALI.

Amino acid sensing in hypothalamic tanycytes via umami taste receptors.

Science.gov (United States)

Lazutkaite, Greta; Soldà, Alice; Lossow, Kristina; Meyerhof, Wolfgang; Dale, Nicholas

2017-11-01

Hypothalamic tanycytes are glial cells that line the wall of the third ventricle and contact the cerebrospinal fluid (CSF). While they are known to detect glucose in the CSF we now show that tanycytes also detect amino acids, important nutrients that signal satiety. Ca 2+ imaging and ATP biosensing were used to detect tanycyte responses to l-amino acids. The downstream pathway of the responses was determined using ATP receptor antagonists and channel blockers. The receptors were characterized using mice lacking the Tas1r1 gene, as well as an mGluR4 receptor antagonist. Amino acids such as Arg, Lys, and Ala evoke Ca 2+ signals in tanycytes and evoke the release of ATP via pannexin 1 and CalHM1, which amplifies the signal via a P2 receptor dependent mechanism. Tanycytes from mice lacking the Tas1r1 gene had diminished responses to lysine and arginine but not alanine. Antagonists of mGluR4 greatly reduced the responses to alanine and lysine. Two receptors previously implicated in taste cells, the Tas1r1/Tas1r3 heterodimer and mGluR4, contribute to the detection of a range of amino acids by tanycytes in CSF. Copyright © 2017 The Authors. Published by Elsevier GmbH.. All rights reserved.

Whole-protein alanine-scanning mutagenesis of allostery: A large percentage of a protein can contribute to mechanism.

Science.gov (United States)

Tang, Qingling; Fenton, Aron W

2017-09-01

Many studies of allosteric mechanisms use limited numbers of mutations to test whether residues play "key" roles. However, if a large percentage of the protein contributes to allosteric function, mutating any residue would have a high probability of modifying allostery. Thus, a predicted mechanism that is dependent on only a few residues could erroneously appear to be supported. We used whole-protein alanine-scanning mutagenesis to determine which amino acid sidechains of human liver pyruvate kinase (hL-PYK; approved symbol PKLR) contribute to regulation by fructose-1,6-bisphosphate (Fru-1,6-BP; activator) and alanine (inhibitor). Each nonalanine/nonglycine residue of hL-PYK was mutated to alanine to generate 431 mutant proteins. Allosteric functions in active proteins were quantified by following substrate affinity over a concentration range of effectors. Results show that different residues contribute to the two allosteric functions. Only a small fraction of mutated residues perturbed inhibition by alanine. In contrast, a large percentage of mutated residues influenced activation by Fru-1,6-BP; inhibition by alanine is not simply the reverse of activation by Fru-1,6-BP. Moreover, the results show that Fru-1,6-BP activation would be extremely difficult to elucidate using a limited number of mutations. Additionally, this large mutational data set will be useful to train and test computational algorithms aiming to predict allosteric mechanisms. © 2017 Wiley Periodicals, Inc.

Amino acid biogeo- and stereochemistry in coastal Chilean sediments

DEFF Research Database (Denmark)

Lomstein, Bente Aagaard; Jørgensen, Bo Barker; Schubert, Carsten J.

2006-01-01

The spatial distribution of total hydrolysable amino acids (THAA) and amino acid enantiomers (D- and L-forms) was investigated in sediments underlying two contrasting Chilean upwelling regions,: at ~23°S off Antofagasta and at ~36°S off Concepcion. The contribution of amino acids to total organic...... carbon (%TAAC: 7-14%) and total nitrogen (%TAAN: 23-38%) in surface sediments decreased with increasing water depth (from 126 to 1350 m) indicating that organic matter becomes increasingly decomposed in surface sediments at greater water depth. Changes in the ratio between the protein amino acid...... aspartate and its non-protein degradation product β-alanine confirmed this observation. Furthermore, estimates of THAA mineralization showed that sedimentary amino acid reactivity decreased with both increasing water depth as well as progressive degradation status of the organic matter that was incorporated...

Protolytic properties and complexation of DL-alpha-alanine and DL-alpha-valine and their dipeptides in aqueous and micellar solutions of surfactants

NARCIS (Netherlands)

Chernyshova, O. S.; Boychenko, Oleksandr; Abdulrahman, H.; Loginova, L. P.

In this work we investigated the effect of the micellar media of anionic (sodium dodecylsulfate, SDS), cationic (cetylpiridinium chloride, CPC) and non-ionic (Brij-35) surfactants on the protolytic properties of amino acids DL-alpha-alanine, DL-alpha-valine and dipeptides

Detecting Electron Transport of Amino Acids by Using Conductance Measurement

Directory of Open Access Journals (Sweden)

Wei-Qiong Li

2017-04-01

Full Text Available The single molecular conductance of amino acids was measured by a scanning tunneling microscope (STM break junction. Conductance measurement of alanine gives out two conductance values at 10−1.85 G0 (1095 nS and 10−3.7 G0 (15.5 nS, while similar conductance values are also observed for aspartic acid and glutamic acid, which have one more carboxylic acid group compared with alanine. This may show that the backbone of NH2–C–COOH is the primary means of electron transport in the molecular junction of aspartic acid and glutamic acid. However, NH2–C–COOH is not the primary means of electron transport in the methionine junction, which may be caused by the strong interaction of the Au–SMe (methyl sulfide bond for the methionine junction. The current work reveals the important role of the anchoring group in the electron transport in different amino acids junctions.

Enantiomer-specific selection of amino acids.

Science.gov (United States)

Ren, Xueying; Tellez, Luis A; de Araujo, Ivan E

2013-12-01

Dietary intake of L-amino acids impacts on several physiological functions, including the control of gastrointestinal motility, pancreatic secretion, and appetite. However, the biological mechanisms regulating behavioral predilections for certain amino acid types remain poorly understood. We tested the hypothesis that, in mice, the potency with which a given glucogenic amino acid increases glucose utilization reflects its rewarding properties. We have found that: (1) during long-, but not short-, term preference tests, L-alanine and L-serine were preferred over their D-enantiomer counterparts, while no such effect was observed for L-threonine vs. D-threonine; (2) these behavioral patterns were closely associated with the ability of L-amino acids to promote increases in respiratory exchange ratios such that those, and only those, L-amino acids able to promote increases in respiratory exchange ratios were preferred over their D-isomers; (3) these behavioral preferences were independent of gustatory influences, since taste-deficient Trpm5 knockout mice displayed ingestive responses very similar to those of their wild-type counterparts. We conclude that the ability to promote increases in respiratory exchange ratios enhances the reward value of nutritionally relevant amino acids and suggest a mechanistic link between substrate utilization and amino acid preferences.

Study on the source of optical activity: Pt. 6. ESR spectroscopy on the asymmetrical radical yields in 90Sr-90Y β irradiated D-and L-alanine

International Nuclear Information System (INIS)

Wang Wenqing; Zhao Jian; Ding Xiang

1993-01-01

Free radical formation in 90 Sr- 90 Y β irradiated D- and L-alanine is studied by ESR spectroscopy. To calibrate the probable differences in the size of ESR tubes, the different densities of alanine and the incidental different impurities, samples are firstly irradiated with 60 Co γ rays. Then the D- and L- alanine samples are irradiated with 90 Sr- 90 Y source at 77 K. Soon after irradiation, ESR measurement is performed on each sample. The average ratio H β+γ /H γ of D- alanine is 0.147 higher than that of L- alanine, indicating that more free radicals are induced in D- alanine by β irradiation. Irradiation at 77 K is capable of diminishing the effect of thermal movement on the polarization of β electrons and freezing the free radical formation. The experiments show the stereoselective interaction of β electrons with D- and L- amino acids, and so supports the Vester-ulbricht hypothesis

Simultaneous determination of F-beta-alanine and beta-alanine in plasma and urine with dual-column reversed-phase high-performance liquid chromatography

NARCIS (Netherlands)

van Kuilenburg, A. B.; Stroomer, A. E.; Peters, G. J.; van Gennip, A. H.

2001-01-01

F-beta-Alanine and beta-alanine were detected in plasma and urine samples with fluorescence detection of orthophthaldialdehyde derivatives of F-beta-alanine and beta-alanine after separation with dual-column reversed-phase HPLC. The detection limits of F-beta-alanine and beta-alanine in the HPLC

Amino Acid Transporters and Release of Hydrophobic Amino Acids in the Heterocyst-Forming Cyanobacterium Anabaena sp. Strain PCC 7120

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Rafael Pernil

2015-04-01

Full Text Available Anabaena sp. strain PCC 7120 is a filamentous cyanobacterium that can use inorganic compounds such as nitrate or ammonium as nitrogen sources. In the absence of combined nitrogen, it can fix N2 in differentiated cells called heterocysts. Anabaena also shows substantial activities of amino acid uptake, and three ABC-type transporters for amino acids have been previously characterized. Seven new loci encoding predicted amino acid transporters were identified in the Anabaena genomic sequence and inactivated. Two of them were involved in amino acid uptake. Locus alr2535-alr2541 encodes the elements of a hydrophobic amino acid ABC-type transporter that is mainly involved in the uptake of glycine. ORF all0342 encodes a putative transporter from the dicarboxylate/amino acid:cation symporter (DAACS family whose inactivation resulted in an increased uptake of a broad range of amino acids. An assay to study amino acid release from Anabaena filaments to the external medium was set up. Net release of the alanine analogue α-aminoisobutyric acid (AIB was observed when transport system N-I (a hydrophobic amino acid ABC-type transporter was engaged in the uptake of a specific substrate. The rate of AIB release was directly proportional to the intracellular AIB concentration, suggesting leakage from the cells by diffusion.

Solid state radiolysis of amino acids in an astrochemical perspective

International Nuclear Information System (INIS)

Cataldo, Franco; Angelini, Giancarlo; Iglesias-Groth, Susana; Manchado, Arturo

2011-01-01

The aliphatic amino acids L-alanine and L-leucine and the aromatic amino acids L-phenylalanine, L-tyrosine and L-tryptophan were irradiated in the solid state to a dose of 3.2 MGy. The degree of decomposition was measured by differential scanning calorimetry (DSC). Furthermore the degree of radioracemization was measured by optical rotatory dispersion (ORD) spectroscopy. From the DSC measurement a radiolysis rate constant k and the half life T 1/2 for each amino acid have been determined and extrapolated to a dose of 14 MGy, which corresponds to the expected total dose delivered by the decay of radionuclides to the organic molecules present in comets and asteroids in 4.6x10 9 years, the age of the Solar System. It is shown that all the amino acids studied can survive a radiation dose of 14 MGy although they are reduced to 1/4-1/5 of their original value they had at the beginning of the history of the Solar System. Consequently, the amount of alanine or leucine found today in the meteorites known as carbonaceous chondrites is just 1/4-1/5 of the amount originally present at the epoch of the formation of the Solar System 4.6x10 9 years ago. Among the amino acids studied, tyrosine shows the highest radiation resistance while tryptophan does not combine its relatively high radiation resistance with an elevated level of radioracemization resistance. Apart from the exception of tryptophan, it is shown that the radiolysis rate constants k of all the amino acids studied are in reasonable agreement with the radioracemization rate constant k rac .

Solid state radiolysis of amino acids in an astrochemical perspective

Energy Technology Data Exchange (ETDEWEB)

Cataldo, Franco, E-mail: franco.cataldo@fastwebnet.i [Istituto Nazionale di Astrofisica-Osservatorio Astrofisica di Catania, Via S. Sofia 78, 95123 Catania (Italy); Lupi Chemical Research, Via Casilina 1626/A, 00133 Rome (Italy); Angelini, Giancarlo [Istituto di Metodologie Chimiche, CNR, Via Salaria Km 29300, 00016 Monterotondo Stazione, Rome (Italy); Iglesias-Groth, Susana [Instituto de Astrofisica de Canarias, Via Lactea s/n, E-38200, La Laguna, Tenerife (Spain); Manchado, Arturo [Instituto de Astrofisica de Canarias, Via Lactea s/n, E-38200, La Laguna, Tenerife (Spain) and CSIC (Spain)

2011-01-15

The aliphatic amino acids L-alanine and L-leucine and the aromatic amino acids L-phenylalanine, L-tyrosine and L-tryptophan were irradiated in the solid state to a dose of 3.2 MGy. The degree of decomposition was measured by differential scanning calorimetry (DSC). Furthermore the degree of radioracemization was measured by optical rotatory dispersion (ORD) spectroscopy. From the DSC measurement a radiolysis rate constant k and the half life T{sub 1/2} for each amino acid have been determined and extrapolated to a dose of 14 MGy, which corresponds to the expected total dose delivered by the decay of radionuclides to the organic molecules present in comets and asteroids in 4.6x10{sup 9} years, the age of the Solar System. It is shown that all the amino acids studied can survive a radiation dose of 14 MGy although they are reduced to 1/4-1/5 of their original value they had at the beginning of the history of the Solar System. Consequently, the amount of alanine or leucine found today in the meteorites known as carbonaceous chondrites is just 1/4-1/5 of the amount originally present at the epoch of the formation of the Solar System 4.6x10{sup 9} years ago. Among the amino acids studied, tyrosine shows the highest radiation resistance while tryptophan does not combine its relatively high radiation resistance with an elevated level of radioracemization resistance. Apart from the exception of tryptophan, it is shown that the radiolysis rate constants k of all the amino acids studied are in reasonable agreement with the radioracemization rate constant k{sub rac}.

Lipidization of Simple and di-Functional Amino Acids

International Nuclear Information System (INIS)

Zainab Idris; Mohd Wahid Samsudin; Salmiah Ahmad

2013-01-01

This paper discuss the modification of azelaic acid into its applicable form by attachment of both its carboxyl sites to N-terminal of amino acid ethyl ester forming amide linkages in anhydrous medium. Acylation of glycine ethyl ester hydrochloride with azelaic acid dichloride was best conducted in a 100 % anhydrous medium. L-amino acid ethyl ester bearing a primary hydroxyl group on its side chain gave mixtures of product and variation in composition depending on the mole ratio of reactants used. Reduction in purity was also observed for L-amino acid ethyl ester with primary -SH group on its side chain as compared to L-amino acid ethyl ester having -SCH 3 group on the L-amino acid side chain. The diamidoester of azelaic acid with L-alanine ethyl ester, L-valine ethyl ester, L-leucine ethyl ester and L-glutamic acid diethyl ester were in good yield when prepared through the modified Schotten-Baumann reaction conditions. (author)

Water-resistant alanine-EPR dosimeter alanpol

International Nuclear Information System (INIS)

Peimel-Stuglik, Zofia; Bryl-Sandelewska, Teresa; Mirkowski, Krzysztof; Sartowska, Bozena

2009-01-01

Alanpol-water-resistant alanine-electron paramagnetic resonance (EPR) dosimeter consisted of cheap DL-α-alanine (9.8-27%) suspended in polyethylene matrix was presented. The rods (O=2.8 mm) were extruded from a hot mixture of alanine and low-density polyethylene. No grinding or crushing was used for alanine preparation. An orientation of cylindrical crystals, up to 300 μm long in parallel to the rod axis was responsible for some differences in a shape of EPR signal. These differences had no negative consequences for dosimetric applications. Signal-to-dose dependence was linear up to 10 kGy. Standard deviation of dosimetric answer was up to ±1.8% and up to 2.4% for dosimeters with 9.8% and 27% of DL-α-alanine, respectively. Irradiation temperature coefficient for both dosimeters was equal 0.2%/ deg. C. Hydrophobic properties of polyethylene and small number of alanine crystals located on the surface of the rod led to high resistance of dosimeters to water and humidity. The 24 h soaking of irradiated dosimeters in liquid water-reduced EPR signals by 3-4% and by 2-3% for dosimeters with 27% and 9.8% of DL-α-alanine, respectively. Three month storage time of irradiated dosimeters in room conditions decreases EPR signal for ∼3%.

Serum Neuron-Specific Enolase, Biogenic Amino-Acids and Neurobehavioral Function in Lead-Exposed Workers from Lead-Acid Battery Manufacturing Process

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K Ravibabu

2015-01-01

Full Text Available Background: The interaction between serum neuron-specific enolase (NSE, biogenic amino-acids and neurobehavioral function with blood lead levels in workers exposed to lead form lead-acid battery manufacturing process was not studied. Objective: To evaluate serum NSE and biogenic amino-acids (dopamine and serotonin levels, and neurobehavioral performance among workers exposed to lead from lead-acid storage battery plant, and its relation with blood lead levels (BLLs. Methods: In a cross-sectional study, we performed biochemical and neurobehavioral function tests on 146 workers exposed to lead from lead-acid battery manufacturing process. BLLs were assessed by an atomic absorption spectrophotometer. Serum NSE, dopamine and serotonin were measured by ELISA. Neurobehavioral functions were assessed by CDC-recommended tests—simple reaction time (SRT, symbol digit substitution test (SDST, and serial digit learning test (SDLT. Results: There was a significant correlation (r 0.199, p<0.05 between SDST and BLL. SDLT and SRT had also a significant positive correlation (r 0.238, p<0.01. NSE had a negative correlation (r –0.194, p<0.05 with serotonin level. Multiple linear regression analysis revealed that both SRT and SDST had positive significant associations with BLL. SRT also had a positive significant association with age. Conclusion: Serum NSE cannot be used as a marker for BLL. The only domain of neurobehavioral function tests that is affected by increased BLL in workers of lead-acid battery manufacturing process is that of the “attention and perception” (SDST.

Formation of amino acids by cobalt-60 irradiation of hydrogen cyanide solutions

Science.gov (United States)

Sweeney, M. A.; Toste, A. P.; Ponnamperuma, C.

1976-01-01

Experiments were conducted to study the pathway for the prebiotic origin of amino acids from hydrogen cyanide (HCN) under the action of ionizing radiation considered as an effective source of energy on the primitive earth. The irradiations were performed in a cobalt-60 source with a dose rate of 200,000 rad/hr. Seven naturally occurring amino acids are identified among the products formed by the hydrolysis of gamma-irradiated solutions of HCN: glycine, alanine, valine, serine, threonine, aspartic acid, and glutamic acid. The identity of these amino acids is established by gas chromatography and mass spectrometry. Control experiments provided evidence that the amino acids are not the result of contamination.

Subcritical Water Extraction of Amino Acids from Atacama Desert Soils

Science.gov (United States)

Amashukeli, Xenia; Pelletier, Christine C.; Kirby, James P.; Grunthaner, Frank J.

2007-01-01

Amino acids are considered organic molecular indicators in the search for extant and extinct life in the Solar System. Extraction of these molecules from a particulate solid matrix, such as Martian regolith, will be critical to their in situ detection and analysis. The goals of this study were to optimize a laboratory amino acid extraction protocol by quantitatively measuring the yields of extracted amino acids as a function of liquid water temperature and sample extraction time and to compare the results to the standard HCl vapor- phase hydrolysis yields for the same soil samples. Soil samples from the Yungay region of the Atacama Desert ( Martian regolith analog) were collected during a field study in the summer of 2005. The amino acids ( alanine, aspartic acid, glutamic acid, glycine, serine, and valine) chosen for analysis were present in the samples at concentrations of 1 - 70 parts- per- billion. Subcritical water extraction efficiency was examined over the temperature range of 30 - 325 degrees C, at pressures of 17.2 or 20.0 MPa, and for water- sample contact equilibration times of 0 - 30 min. None of the amino acids were extracted in detectable amounts at 30 degrees C ( at 17.2 MPa), suggesting that amino acids are too strongly bound by the soil matrix to be extracted at such a low temperature. Between 150 degrees C and 250 degrees C ( at 17.2 MPa), the extraction efficiencies of glycine, alanine, and valine were observed to increase with increasing water temperature, consistent with higher solubility at higher temperatures, perhaps due to the decreasing dielectric constant of water. Amino acids were not detected in extracts collected at 325 degrees C ( at 20.0 MPa), probably due to amino acid decomposition at this temperature. The optimal subcritical water extraction conditions for these amino acids from Atacama Desert soils were achieved at 200 degrees C, 17.2 MPa, and a water- sample contact equilibration time of 10 min.

Amino Acids Content in Mangosteen Fruit as Affected by Tree Ages

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Eko Setiawan

2018-01-01

Full Text Available Kita meneliti perbedaan kandungan dan komposisi asam amino pada buah manggis yang dipanen dari pohon yang berbeda umur dari kebun Manggis di Bogor pada tahun 2012. Pohon manggis dengan umur yang berbeda (muda, sekitar 20 tahun; sedang, sekitar umur 35 tahun; dan tua, sekitar 50 tahundipilih untuk diambil buahnya. Buah dari tiap pohon dipanen pada kondisi masak hijau spot kuning-merah. Pada buah dari semua umur pohon, asam amino yang dominan adalah γ-aminobutyric acid dan alanine. Total kandungan asam amino tertinggi terdapat pada buah yang dipetik dari pohon umur muda dari pada  buah yang dipetik dari pohon umur sedang dan tuaKata kunci: asam amino, GABA, kandungan, manggis

Toxicological Influence of Dietary Fumonisin B, on Blood Profile of ...

African Journals Online (AJOL)

Rabbits exposed to 7.5 and 10.0 mg/kg FB1 showed a hypercholesterolemia condition. Activities of aspartate amino transferase, alanine amino transferase and alkaline phosphatase were significantly (P<0.05) elevated in the serum of treated animals, which is an indication of liver and kidney toxicity and cell damage.

Alanine-polymer dosemeter of ionizing radiation

International Nuclear Information System (INIS)

Tomasinski, Z.; Mirkowski, K.; Panta, P.; Stachowicz, W.

1994-01-01

The method of chemical preparation of alanine-copolymer of ethylene and vinyl acetate has been worked out. The material has been in a form of rods. The content of alanine has not exceeded 30%. The ESR signal of alanine radicals has been detected after exposition to ionizing radiation. The dose-response relationship has been presented

Clinical implications in the prevalence and associated cardiovascular factors of elevated serum alanine aminotransferase levels among elderly agricultural and fishing population in Taipei, Taiwan: experience at a teaching hospital.

Science.gov (United States)

Chen, Yu-Fen; Hu, Yi-Chun; Shen, Hsi-Che; Chang, Hui-Te; Tung, Tao-Hsin

2014-01-01

To discuss the prevalence and associated factors related to an elevated serum alanine aminotransferase (ALT) level among the elderly agricultural and fishing population. A total of 6542 (3989 males and 2553 females) healthy adults voluntarily admitted to a teaching hospital for a physical checkup in 2010 in Taipei, Taiwan. Fasting blood samples were drawn via venipuncture, and clinical nurses interviewed the study participants using a structured questionnaire from. The overall prevalence of an elevated serum ALT level was 18.2% and revealed a statistically significant decrease with increasing age (P < 0.001). The men exhibited a higher prevalence than the women (19.7% vs 15.9%; P < 0.001). Male sex; younger age; and presence of obesity, hypertension, hyperuricemia, and hypoalbuminemia were significantly associated with an elevated serum ALT level. Sex-related differences were also revealed. For the men, type 2 diabetes (odds ratio [OR], 1.23; 95% confidence interval [CI], 1.02-1.57), hypercholesterolemia (OR, 1.78; 95% CI, 1.22-2.83), hypertriglyceridemia (OR, 1.32; 95% CI, 1.04-1.73), and low high-density lipoprotein (OR, 1.26; 95% CI, 1.05-1.51) were significantly related to an elevated serum ALT level, but this was not so for the women. The disparity of ALT in age groups was revealed. Several sex-related differences were indicated pertaining to the prevalence of an elevated serum ALT level among elderly specific occupational population.

Protein and amino acid metabolism in skeletal muscle

Energy Technology Data Exchange (ETDEWEB)

Wu, Guoyao.

1989-01-01

Isolated chick extensor digitorum communis (EDC) muscles and, in some experiments, rat skeletal muscles were used to study a number of aspects of protein and amino acid metabolism. (1) Chick EDC muscles synthesize and release large amounts of alanine and glutamine, which indirectly obtain their amino groups from branched-chain amino acids (BCAA). (2) Acetoacetate or DL-{beta}-hydroxybutyrate (4 mM) decrease (P < 0.01) alanine synthesis and BCAA transamination in EDC muscles from 24-h fasted chicks by decreasing (P < 0.01) intracellular concentrations of pyruvate due to inhibition of glycolysis. (3) Glutamine is extensively degraded in skeletal muscles from both chicks and rats, thus challenging the traditional view that glutamine oxidation is negligible in skeletal muscle. The cytosolic glutamine aminotransferases L and K in the rat and the mitochondrial phosphate-activated glutaminase in the chick play important roles in the conversion of glutamine to {alpha}-ketoglutarate for further oxidation. (4) Although methionine has been reported to be extensively transaminated in rat skeletal muscle preparations in the absence of other amino acids, transamination of methionine is absent or negligible in chick and rat skeletal muscles in the presence of physiological concentrations of amino acids. (5) Glutamine at 1.0-15 mM increases (P < 0.01) protein synthesis ({sup 3}H-phenylalanine incorporation), and at 10.0-15.0 mM decreases (P < 0.05) protein degradation ({sup 3}H-phenylalanine release from prelabelled protein in vivo) in EDC muscles from fed chicks as compared to muscles incubated in the absence of glutamine. (6) Acetoacetate or DL-{beta}-hydroxybutyrate (4 mM) has a small but significant inhibitory effect (P < 0.05) on the rate of protein synthesis, but has no effect (P > 0.05) on the rate of protein degradation in EDC muscles from fed chicks.

Diverse taxa of cyanobacteria produce beta-N-methylamino-L-alanine, a neurotoxic amino acid.

Science.gov (United States)

Cox, Paul Alan; Banack, Sandra Anne; Murch, Susan J; Rasmussen, Ulla; Tien, Georgia; Bidigare, Robert Richard; Metcalf, James S; Morrison, Louise F; Codd, Geoffrey A; Bergman, Birgitta

2005-04-05

Cyanobacteria can generate molecules hazardous to human health, but production of the known cyanotoxins is taxonomically sporadic. For example, members of a few genera produce hepatotoxic microcystins, whereas production of hepatotoxic nodularins appears to be limited to a single genus. Production of known neurotoxins has also been considered phylogenetically unpredictable. We report here that a single neurotoxin, beta-N-methylamino-L-alanine, may be produced by all known groups of cyanobacteria, including cyanobacterial symbionts and free-living cyanobacteria. The ubiquity of cyanobacteria in terrestrial, as well as freshwater, brackish, and marine environments, suggests a potential for wide-spread human exposure.

Diverse taxa of cyanobacteria produce β-N-methylamino-l-alanine, a neurotoxic amino acid

Science.gov (United States)

Cox, Paul Alan; Banack, Sandra Anne; Murch, Susan J.; Rasmussen, Ulla; Tien, Georgia; Bidigare, Robert Richard; Metcalf, James S.; Morrison, Louise F.; Codd, Geoffrey A.; Bergman, Birgitta

2005-01-01

Cyanobacteria can generate molecules hazardous to human health, but production of the known cyanotoxins is taxonomically sporadic. For example, members of a few genera produce hepatotoxic microcystins, whereas production of hepatotoxic nodularins appears to be limited to a single genus. Production of known neurotoxins has also been considered phylogenetically unpredictable. We report here that a single neurotoxin, β-N-methylamino-l-alanine, may be produced by all known groups of cyanobacteria, including cyanobacterial symbionts and free-living cyanobacteria. The ubiquity of cyanobacteria in terrestrial, as well as freshwater, brackish, and marine environments, suggests a potential for wide-spread human exposure. PMID:15809446

Alanine transaminase (ALT) blood test

Science.gov (United States)

... gov/ency/article/003473.htm Alanine transaminase (ALT) blood test To use the sharing features on this page, please enable JavaScript. The alanine transaminase (ALT) blood test measures the level of the enzyme ALT in ...

Elevational Variation in Soil Amino Acid and Inorganic Nitrogen Concentrations in Taibai Mountain, China.

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Xiaochuang Cao

Full Text Available Amino acids are important sources of soil organic nitrogen (N, which is essential for plant nutrition, but detailed information about which amino acids predominant and whether amino acid composition varies with elevation is lacking. In this study, we hypothesized that the concentrations of amino acids in soil would increase and their composition would vary along the elevational gradient of Taibai Mountain, as plant-derived organic matter accumulated and N mineralization and microbial immobilization of amino acids slowed with reduced soil temperature. Results showed that the concentrations of soil extractable total N, extractable organic N and amino acids significantly increased with elevation due to the accumulation of soil organic matter and the greater N content. Soil extractable organic N concentration was significantly greater than that of the extractable inorganic N (NO3--N + NH4+-N. On average, soil adsorbed amino acid concentration was approximately 5-fold greater than that of the free amino acids, which indicates that adsorbed amino acids extracted with the strong salt solution likely represent a potential source for the replenishment of free amino acids. We found no appreciable evidence to suggest that amino acids with simple molecular structure were dominant at low elevations, whereas amino acids with high molecular weight and complex aromatic structure dominated the high elevations. Across the elevational gradient, the amino acid pool was dominated by alanine, aspartic acid, glycine, glutamic acid, histidine, serine and threonine. These seven amino acids accounted for approximately 68.9% of the total hydrolyzable amino acid pool. The proportions of isoleucine, tyrosine and methionine varied with elevation, while soil major amino acid composition (including alanine, arginine, aspartic acid, glycine, histidine, leucine, phenylalanine, serine, threonine and valine did not vary appreciably with elevation (p>0.10. The compositional

Development of fast disintegrating compressed tablets using amino acid as disintegration accelerator: evaluation of wetting and disintegration of tablet on the basis of surface free energy.

Science.gov (United States)

Fukami, Jinichi; Ozawa, Asuka; Yoshihashi, Yasuo; Yonemochi, Etsuo; Terada, Katsuhide

2005-12-01

A fast disintegrating compressed tablet was formulated using amino acids, such as L-lysine HCl, L-alanine, glycine and L-tyrosine as disintegration accelerator. The tablets having the hardness of about 4 kgf were prepared and the effect of amino acids on the wetting time and disintegration time in the oral cavity of tablets was examined on the basis of surface free energy of amino acids. The wetting time of the tablets increased in the order of L-lysine HCl, L-alanine, glycine and L-tyrosine, whereas the disintegration time in the oral cavity of the tablets increased in the order of L-alanine, glycine, L-lysine HCl and L-tyrosine. These behaviors were well analyzed by the introduction of surface free energy. When the polar component of amino acid was large value or the dispersion component was small value, faster wetting of tablet was observed. When the dispersion component of amino acid was large value or the dispersion component was small value, faster disintegration of tablet was observed, expect of L-tyrosine tablet. The fast disintegration of tablets was explained by the theory presented by Matsumaru.

Alanine metabolism in pyridoxine-depleted rat liver

International Nuclear Information System (INIS)

Okada, Mitsuko; Abe, Midori

1976-01-01

Alanine metabolism in normal and pyridoxine-deficient rats was studied in vivo and in vitro. Incorporation of 14 C-alanine into various liver components was determined and no difference was shown between normal and deficient animals in the incorporation into liver homogenates, lipid, protein and plasma glucose. Using the liver slice system, gluconeogenic activity from alanine or pyruvate was 40% lower in deficient rats compared with the activity of normal rats. However, inhibition was completely removed by the addition of 2-oxoglutarate to alanine. Penicillamine did not affect glucose formation from alanine in the liver slice. (auth.)

Crystal Structures of Active Fully Assembled Substrate- and Product-Bound Complexes of UDP-N-Acetylmuramic Acid:l-Alanine Ligase (MurC) from Haemophilus influenzae

OpenAIRE

Mol, Clifford D.; Brooun, Alexei; Dougan, Douglas R.; Hilgers, Mark T.; Tari, Leslie W.; Wijnands, Robert A.; Knuth, Mark W.; McRee, Duncan E.; Swanson, Ronald V.

2003-01-01

UDP-N-acetylmuramic acid:l-alanine ligase (MurC) catalyzes the addition of the first amino acid to the cytoplasmic precursor of the bacterial cell wall peptidoglycan. The crystal structures of Haemophilus influenzae MurC in complex with its substrate UDP-N-acetylmuramic acid (UNAM) and Mg2+ and of a fully assembled MurC complex with its product UDP-N-acetylmuramoyl-l-alanine (UMA), the nonhydrolyzable ATP analogue AMPPNP, and Mn2+ have been determined to 1.85- and 1.7-Å resolution, respective...

An SFG study of interfacial amino acids at the hydrophilic SiO2 and hydrophobic deuterated polystyrene surfaces.

Science.gov (United States)

Holinga, George J; York, Roger L; Onorato, Robert M; Thompson, Christopher M; Webb, Nic E; Yoon, Alfred P; Somorjai, Gabor A

2011-04-27

Sum frequency generation (SFG) vibrational spectroscopy was employed to characterize the interfacial structure of eight individual amino acids--L-phenylalanine, L-leucine, glycine, L-lysine, L-arginine, L-cysteine, L-alanine, and L-proline--in aqueous solution adsorbed at model hydrophilic and hydrophobic surfaces. Specifically, SFG vibrational spectra were obtained for the amino acids at the solid-liquid interface between both hydrophobic d(8)-polystyrene (d(8)-PS) and SiO(2) model surfaces and phosphate buffered saline (PBS) at pH 7.4. At the hydrophobic d(8)-PS surface, seven of the amino acids solutions investigated showed clear and identifiable C-H vibrational modes, with the exception being l-alanine. In the SFG spectra obtained at the hydrophilic SiO(2) surface, no C-H vibrational modes were observed from any of the amino acids studied. However, it was confirmed by quartz crystal microbalance that amino acids do adsorb to the SiO(2) interface, and the amino acid solutions were found to have a detectable and widely varying influence on the magnitude of SFG signal from water at the SiO(2)/PBS interface. This study provides the first known SFG spectra of several individual amino acids in aqueous solution at the solid-liquid interface and under physiological conditions.

Compositional changes of proteins and amino acids in germinating coffee seeds

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Milton Massao Shimizu

2000-01-01

Full Text Available Endosperm is the main reserve tissue in coffee seeds. Coffee (Coffea arabica L. seeds were germinated for six weeks and qualitative and quantitative changes in amino acids and proteins were investigated. The total content of free amino acids were reduced during germination, however, protein content remained constant. SDS-PAGE profiles showed that legumin-like proteins became less stained in the last weeks. Asparagine, glutamic acid, aspartic acid, alanine and lysine were the major free amino acids, although serine and glutamine were also significant. Except for tyrosine, which increased with germination, all other amino acids were reduced. Analysis of the amino acid composition of the total soluble protein showed glutamic acid/glutamine and glycine as the main amino acids. However, other amino acids such as leucine, aspartic acid/asparagine, alanine, lysine, serine were also found in reasonable amounts.Endosperma é o principal tecido de reserva em sementes de café. Sementes de café (Coffea arabica L. foram germinadas por seis semanas e as alterações qualitativas e quantitativas de aminoácidos e proteínas foram investigadas. O conteúdo total de aminoácidos livres reduziu durante a germinação, no entanto, o conteúdo de proteínas permaneceu constante. Perfis eletroforéticos de proteínas em SDS-PAGE mostraram que proteínas do tipo legumina foram menos coradas nas últimas semanas. Asparagina, ácido glutâmico, ácido aspártico, alanina e lisina foram os principais aminoácidos, apesar de que serina e glutamina também estavam presentes em quantidades significativas. Exceto tirosina, a qual aumentou durante a germinação, todos os outros aminoácidos tiveram redução em sua concentração. A análise aminoacídica da fração de proteína solúvel total mostrou que ácido glutâmico/glutamina e glicina eram os principais aminoácidos presentes. No entanto, outros aminoácidos, tais como leucina, ácido asp

Protection against hyperthermic cell killing by alanine

International Nuclear Information System (INIS)

Cunningham, A.; Henle, K.J.; Moss, A.J.; Nagle, W.A.

1987-01-01

Compounds capable of protecting cells against hyperthermia may provide new insights into potential mechanisms of thermotolerance and cellular heat death. The authors characterized heat protection by alanine and related compounds as a function of concentration, temperature and preincubation time. Alanine was added either to complete medium or to HBSS before hyperthermia. Maximal heat protection required 3 hr, 37 0 ; longer preincubation intervals resulted in lower levels of protection. Addition of alanine to medium after hyperthermia had no protective effect. Protection was concentration dependent with a 20- or 200-fold increase in cell survival after 40 min, 45 0 C at 60 mM in medium or in HBSS, respectively. Higher alanine concentrations up to 120mM did not significantly increase heat protection. A 45 0 -heat survival curve showed that 100mM alanine increased the D/sub q/ by approx. 12 min with little change in the D/sub o/. Hyperthermia of 1 hr at temperatures between 42 0 and 45 0 indicated that 100mM alanine shifted the isotoxic temperature by 0.5 Celsius degrees. Polymers of either L or D,L alanine and related compounds, like pyruvate, also protected cells against heat killing. These results indicate that heat protection by alanine shows characteristics that are not shared by polyhydroxy compounds

[Effect of Electroacupuncture at "Neiguan"(PC 6) on Serum and Myocardial Metabolites in Rats with Myocardial Ischemia Reperfusion Injury Based on Nuclear Magnetic Resonance Spectroscopy].

Science.gov (United States)

Tang, Ya-Ni; Tan, Cheng-Fu; Liu, Wei-Wei; Yan, Jie; Wang, Chao; Liu, Mi; Lin, Dong-Hai; Huang, Cai-Hua; Du, Lin; Chen, Mei-Lin; Li, Jiao-Lan; Zhu, Ding-Ming

2018-03-25

We have repeatedly demonstrated that electroacupuncture (EA) of "Neiguan"(PC 6) can improve myocardial ischemia in rats. The present study was designed to investigate the metabolomic profile of peripheral blood se-rum and myocardium involving EA-induced improvement of myocardial ischemia-reperfusion injury (MIRI) in rats by using nuclear magnetic resonance spectroscopy. Thirty male SD rats were equally randomized into blank control, model and EA groups. Rats of the control group were only banded for 20 min, once a day for 7 days. The MIRI model was established by occlusion of the anterior descending branch of the left coronary artery for 40 min, followed by reperfusion for 60 min, and rats of the model group were banded as those in the control group. EA (10 Hz/50 Hz, 1 mA) was applied to bilateral PC 6 for 20 min, once daily for 7 days. The blood samples and left ventricular myocardial tissues were collected for assaying the profiles of differential metabolites using 1 H nuclear magnetic resonance ( 1 H NMR) spectroscopy and multivariate statistical analysis such as the principal components analysis (PCA), partial least squares-discriminant analysis (PLS-DA) and orthogonal PLS-DA (O-PLS-DA) with SIMCA-P software 12.0. A total of 19 differential metabolites (17 down-regulated, 2 up-regulated) in the serum and 14 differential metabolites (13 down-regulated and 1 up-regulated) in the ischemic left myocardium were identified after MIRI. Of the 19 serum differential metabolites, amino acids (leucine, isoleucine, valine,alanine, lysine, glycine, glutamine), 3-hydroxy butyric acid (3-HB), lactic acid, acetate, N-acetyl glycoprotein (NAc), acetone, acetoacetate, succinate, polyunsaturated fatty acids (PUFA), creatine, glycerophosphocholine (GPC) were down-regulated; while low density lipoprotein (LDL), LDL/very low density lipoprotein(LDL/VLDL)and glucose obviously up-regulated. Of the 14 myocardial differential metabolites, amino acids (alanine, lysine, glutamate

Surface Propensity of Atmospherically Relevant Amino Acids Studied by XPS.

Science.gov (United States)

Mocellin, Alexandra; Gomes, Anderson Herbert de Abreu; Araújo, Oscar Cardoso; de Brito, Arnaldo Naves; Björneholm, Olle

2017-04-27

Amino acids constitute an important fraction of the water-soluble organic nitrogen (WSON) compounds in aerosols and are involved in many processes in the atmosphere. In this work, we applied X-ray photoelectron spectroscopy (XPS) to study aqueous solutions of four amino acids, glycine, alanine, valine, and methionine, in their zwitterionic forms. We found that amino acids with hydrophilic side chains and smaller size, GLY and ALA, tend to stay in the bulk of the liquid, while the hydrophobic and bigger amino acids, VAL and MET, are found to concentrate more on the surface. We found experimental evidence that the amino acids have preferential orientation relative to the surface, with the hydrophobic side chain being closer to the surface than the hydrophilic carboxylate group. The observed amino acid surface propensity has implications in atmospheric science as the surface interactions play a central role in cloud droplet formation, and they should be considered in climate models.

Carbobenzoxy amino acids: Structural requirements for cholecystokinin receptor antagonist activity

International Nuclear Information System (INIS)

Maton, P.N.; Sutliff, V.E.; Jensen, R.T.; Gardner, J.D.

1985-01-01

The authors used dispersed acini prepared from guinea pig pancreas to examine 28 carbobenzoxy (CBZ) amino acids for their abilities to function as cholecystokinin receptor antagonists. All amino acid derivatives tested, except for CBZ-alanine, CBZ-glycine, and N alpha-CBZ- lysine, were able to inhibit the stimulation of amylase secretion caused by the C-terminal octapeptide of cholecystokinin. In general, there was a good correlation between the ability of a carbobenzoxy amino acid to inhibit stimulated amylase secretion and the ability of the amino acid derivative to inhibit binding of 125 I-cholecystokinin. The inhibition of cholecystokinin-stimulated amylase secretion was competitive, fully reversible, and specific for those secretagogues that interact with the cholecystokinin receptor. The potencies with which the various carbobenzoxy amino acids inhibited the action of cholecystokinin varied 100-fold and CBZ-cystine was the most potent cholecystokinin receptor antagonist. This variation in potency was primarily but not exclusively a function of the hydrophobicity of the amino acid side chain

Isotopic effects in mechanistic studies of biotransformations of fluorine derivatives of L-alanine catalysed by L-alanine dehydrogenase

International Nuclear Information System (INIS)

Szymańska-Majchrzak, Jolanta; Pałka, Katarzyna; Kańska, Marianna

2017-01-01

Synthesis of 3-fluoro-[2- 2 H]-L-alanine (3-F-[ 2 H]-L-Ala) in reductive amination of 3-fluoropyruvic acid catalysed by L-alanine dehydrogenase (AlaDH) was described. Fluorine derivative was used to study oxidative deamination catalysed by AlaDH applied kinetic (for 3-F-L-Ala in H 2 O - KIE’s on V max : 1.1; on V max /K M : 1.2; for 3-F-L-Ala in 2 H 2 O – on V max : 1.4; on V max /K M : 2.1) and solvent isotope effect methods (for 3-F-L-Ala - SIE’s on V max : 1.0; on V max /K M : 0.87; for 3-F-[2- 2 H]-L-Ala – on V max : 1.4; on V max /K M : 1.5). Studies explain some details of reaction mechanism. - Highlights: • Synthesis of 3-fluoro-[2- 2 H]-L-alanine was performed. • The reactions were catalysed using the enzyme L-alanine dehydrogenase. • Performed reactions involved fluorinated analogues of L-alanine. • Solvent isotope effects of deuterium were determined. • Kinetic isotope effects were determined for obtained 3-fluoro-L-alanine. • The mechanism of reaction catalysed by L-alanine dehydrogenase was proposed.

Simultaneous quantification of arginine, alanine, methionine and cysteine amino acids in supplements using a novel bioelectro-nanosensor based on CdSe quantum dot/modified carbon nanotube hollow fiber pencil graphite electrode via Taguchi method.

Science.gov (United States)

Hooshmand, Sara; Es'haghi, Zarrin

2017-11-30

A number of four amino acids have been simultaneously determined at CdSe quantum dot-modified/multi-walled carbon nanotube hollow fiber pencil graphite electrode in different bodybuilding supplements. CdSe quantum dots were synthesized and applied to construct a modified carbon nanotube hollow fiber pencil graphite electrode. FT-IR, TEM, XRD and EDAX methods were applied for characterization of the synthesized CdSe QDs. The electro-oxidation of arginine (Arg), alanine (Ala), methionine (Met) and cysteine (Cys) at the surface of the modified electrode was studied. Then the Taguchi's method was applied using MINITAB 17 software to find out the optimum conditions for the amino acids determination. Under the optimized conditions, the differential pulse (DP) voltammetric peak currents of Arg, Ala, Met and Cys increased linearly with their concentrations in the ranges of 0.287-33670μM and detection limits of 0.081, 0.158, 0.094 and 0.116μM were obtained for them, respectively. Satisfactory results were achieved for calibration and validation sets. The prepared modified electrode represents a very good resolution between the voltammetric peaks of the four amino acids which makes it suitable for the detection of each in presence of others in real samples. Copyright © 2017. Published by Elsevier B.V.

Crystallization and preliminary crystallographic analysis of d-alanine-d-alanine ligase from Streptococcus mutans

Energy Technology Data Exchange (ETDEWEB)

Lu, Yong-Zhi; Sheng, Yu [Institute for Nanobiomedical Technology and Membrane Biology, West China Hospital, Sichuan University, Chengdu 610065, Sichuan (China); Li, Lan-Fen [National Laboratory of Protein Engineering and Plant Genetic Engineering, College of Life Sciences, Peking University, Beijing 100871 (China); Tang, De-Wei [Institute for Nanobiomedical Technology and Membrane Biology, West China Hospital, Sichuan University, Chengdu 610065, Sichuan (China); Liu, Xiang-Yu [National Laboratory of Protein Engineering and Plant Genetic Engineering, College of Life Sciences, Peking University, Beijing 100871 (China); Zhao, Xiaojun, E-mail: zhaoxj@scu.edu.cn [Institute for Nanobiomedical Technology and Membrane Biology, West China Hospital, Sichuan University, Chengdu 610065, Sichuan (China); Liang, Yu-He, E-mail: zhaoxj@scu.edu.cn; Su, Xiao-Dong [National Laboratory of Protein Engineering and Plant Genetic Engineering, College of Life Sciences, Peking University, Beijing 100871 (China); Institute for Nanobiomedical Technology and Membrane Biology, West China Hospital, Sichuan University, Chengdu 610065, Sichuan (China)

2007-09-01

A potential target for antibiotic drug design, d-alanine-d-alanine ligase from S. mutans, was expressed in E. coli, purified and crystallized. Diffraction data were collected to 2.4 Å resolution. d-Alanine-d-alanine ligase is encoded by the gene ddl (SMU-599) in Streptococcus mutans. This ligase plays a very important role in cell-wall biosynthesis and may be a potential target for drug design. To study the structure and function of this ligase, the gene ddl was amplified from S. mutans genomic DNA and cloned into the expression vector pET28a. The protein was expressed in soluble form in Escherichia coli strain BL21 (DE3). Homogeneous protein was obtained using a two-step procedure consisting of Ni{sup 2+}-chelating and size-exclusion chromatography. Purified protein was crystallized and the cube-shaped crystal diffracted to 2.4 Å. The crystal belongs to space group P3{sub 1}21 or P3{sub 2}21, with unit-cell parameters a = b = 79.50, c = 108.97 Å. There is one molecule per asymmetric unit.

Crystallization and preliminary crystallographic analysis of d-alanine-d-alanine ligase from Streptococcus mutans

International Nuclear Information System (INIS)

Lu, Yong-Zhi; Sheng, Yu; Li, Lan-Fen; Tang, De-Wei; Liu, Xiang-Yu; Zhao, Xiaojun; Liang, Yu-He; Su, Xiao-Dong

2007-01-01

A potential target for antibiotic drug design, d-alanine-d-alanine ligase from S. mutans, was expressed in E. coli, purified and crystallized. Diffraction data were collected to 2.4 Å resolution. d-Alanine-d-alanine ligase is encoded by the gene ddl (SMU-599) in Streptococcus mutans. This ligase plays a very important role in cell-wall biosynthesis and may be a potential target for drug design. To study the structure and function of this ligase, the gene ddl was amplified from S. mutans genomic DNA and cloned into the expression vector pET28a. The protein was expressed in soluble form in Escherichia coli strain BL21 (DE3). Homogeneous protein was obtained using a two-step procedure consisting of Ni 2+ -chelating and size-exclusion chromatography. Purified protein was crystallized and the cube-shaped crystal diffracted to 2.4 Å. The crystal belongs to space group P3 1 21 or P3 2 21, with unit-cell parameters a = b = 79.50, c = 108.97 Å. There is one molecule per asymmetric unit

Thermodynamics of DL-alanine solvation in water-dimethylsulfoxide mixtures at 298.15 K

Science.gov (United States)

Roy, S.; Mahali, K.; Mondal, S.; Dolui, B. K.

2015-04-01

In this study we mainly discuss the transfer Gibbs free energy Δ G {/t 0}( i) and Δ S {/t 0}( i)entropy of DL-alanine at 298.15 K and consequently the involved chemical transfer free energy (Δ G {/t,ch 0}( i)) and entropy ( TΔ S {/t,ch 0}( i)) in aqueous mixtures of dimethylsulfoxide are discussed to clarify the solvation chemistry of DL-alanine. For the evaluation of these energy terms, solubility of this amino acid has been measured by formol titrimetry at five equidistant temperatures i.e., from 288.15 to 308.15 K in different composition of this mixed solvent system. The various solvent parameters as well as thermodynamic parameters like molar volume, density, dipole moment and solvent diameter of this solvent system have also been reported here. The chemical effects of the transfer Gibbs energies (Δ G {/t,ch 0}( i)) and entropies of transfer ( TΔ S {/t,ch 0}( i)) have been obtained after elimination of cavity effect and dipole-dipole interaction effects from the total transfer energies. Here the chemical contribution of transfer energetics of DL-alanine is mainly guided by the composite effects of increased dispersion interaction, basicity effect and decreased acidity, hydrogen bonding effects, hydrophilic hydration and hydrophobic hydration of aqueous DMSO mixtures as compared to that of reference solvent, water.

Amino acid profiles of young adults differ by sex, body mass index and insulin resistance.

Science.gov (United States)

Guevara-Cruz, M; Vargas-Morales, J M; Méndez-García, A L; López-Barradas, A M; Granados-Portillo, O; Ordaz-Nava, G; Rocha-Viggiano, A K; Gutierrez-Leyte, C A; Medina-Cerda, E; Rosado, J L; Morales, J C; Torres, N; Tovar, A R; Noriega, L G

2018-04-01

An increase in plasma branched-chain amino acids is associated with a higher risk of developing type 2 diabetes and cardiovascular diseases. However, little is known about the basal plasma amino acid concentrations in young adults. Our aim was to determine the plasma amino acid profiles of young adults and to evaluate how these profiles were modified by sex, body mass index (BMI) and insulin resistance (IR). We performed a transversal study with 608 Mexican young adults aged 19.9 ± 2.4 years who were applicants to the Universidad Autónoma de San Luis Potosí. The subjects underwent a physical examination and provided a clinical history and a blood sample for biochemical, hormonal and amino acid analyses. The women had higher levels of arginine, aspartate and serine and lower levels of α-aminoadipic acid, cysteine, isoleucine, leucine, methionine, proline, tryptophan, tyrosine, urea and valine than the men. The obese subjects had higher levels of alanine, aspartate, cysteine, ornithine, phenylalanine, proline and tyrosine and lower levels of glycine, ornithine and serine than the normal weight subjects. Subjects with IR (defined as HOMA > 2.5) had higher levels of arginine, alanine, aspartate, isoleucine, leucine, phenylalanine, proline, tyrosine, taurine and valine than the subjects without IR. Furthermore, we identified two main groups in the subjects with obesity and/or IR; one group was composed of amino acids that positively correlated with the clinical, biochemical and hormonal parameters, whereas the second group exhibited negative correlations. This study demonstrates that young adults with obesity or IR have altered amino acid profiles characterized by an increase in alanine, aspartate, proline and tyrosine and a decrease in glycine. Copyright © 2018 The Italian Society of Diabetology, the Italian Society for the Study of Atherosclerosis, the Italian Society of Human Nutrition, and the Department of Clinical Medicine and Surgery, Federico II

Cellular and Physiological Effects of Dietary Supplementation with β-Hydroxy-β-Methylbutyrate (HMB and β-Alanine in Late Middle-Aged Mice.

Directory of Open Access Journals (Sweden)

Julian Vallejo

Full Text Available There is growing evidence that severe decline of skeletal muscle mass and function with age may be mitigated by exercise and dietary supplementation with protein and amino acid ingredient technologies. The purposes of this study were to examine the effects of the leucine catabolite, beta-hydroxy-beta-methylbutyrate (HMB, in C2C12 myoblasts and myotubes, and to investigate the effects of dietary supplementation with HMB, the amino acid β-alanine and the combination thereof, on muscle contractility in a preclinical model of pre-sarcopenia. In C2C12 myotubes, HMB enhanced sarcoplasmic reticulum (SR calcium release beyond vehicle control in the presence of all SR agonists tested (KCl, P<0.01; caffeine, P = 0.03; ionomycin, P = 0.03. HMB also improved C2C12 myoblast viability (25 μM HMB, P = 0.03 and increased proliferation (25 μM HMB, P = 0.04; 125 μM HMB, P<0.01. Furthermore, an ex vivo muscle contractility study was performed on EDL and soleus muscle from 19 month old, male C57BL/6nTac mice. For 8 weeks, mice were fed control AIN-93M diet, diet with HMB, diet with β-alanine, or diet with HMB and β-alanine. In β-alanine fed mice, EDL muscle showed a 7% increase in maximum absolute force compared to the control diet (202 ± 3vs. 188± 5 mN, P = 0.02. At submaximal frequency of stimulation (20 Hz, EDL from mice fed HMB plus β-alanine showed an 11% increase in absolute force (88.6 ± 2.2 vs. 79.8 ± 2.4 mN, P = 0.025 and a 13% increase in specific force (12.2 ± 0.4 vs. 10.8 ± 0.4 N/cm2, P = 0.021. Also in EDL muscle, β-alanine increased the rate of force development at all frequencies tested (P<0.025, while HMB reduced the time to reach peak contractile force (TTP, with a significant effect at 80 Hz (P = 0.0156. In soleus muscle, all experimental diets were associated with a decrease in TTP, compared to control diet. Our findings highlight beneficial effects of HMB and β-alanine supplementation on skeletal muscle function in aging mice.

Covalently functionalized graphene sheets with biocompatible natural amino acids

International Nuclear Information System (INIS)

Mallakpour, Shadpour; Abdolmaleki, Amir; Borandeh, Sedigheh

2014-01-01

Graphene sheets were covalently functionalized with aromatic–aliphatic amino acids (phenylalanine and tyrosine) and aliphatic amino acids (alanine, isoleucine, leucine, methionine and valine) by simple and green procedure. For this aim, at first natural graphite was converted into graphene oxide (GO) through strong oxidation procedure; then, based on the surface-exposed epoxy and carboxylic acid groups in GO solid, its surface modification with naturally occurring amino acids, occurred easily throughout the corresponding nucleophilic substitution and condensation reactions. Amino acid functionalized graphene demonstrates stable dispersion in water and common organic solvents. Fourier transform infrared, Raman and X-ray photoelectron spectroscopies, X-ray diffraction, field emission scanning electron microscopy and transmission electron microscopy were used to investigate the nanostructures and properties of prepared materials. Each amino acid has different considerable effects on the structure and morphology of the pure graphite, from increasing the layer spacing to layer scrolling, based on their structures, functional groups and chain length. In addition, therogravimetric analysis was used for demonstrating a successful grafting of amino acid molecules to the surface of graphene.

Potential transfer of neurotoxic amino acid β-N-methylamino-alanine (BMAA) from mother to infant during breast-feeding: Predictions from human cell lines.

Science.gov (United States)

Andersson, Marie; Ersson, Lisa; Brandt, Ingvar; Bergström, Ulrika

2017-04-01

β-N-methylamino-alanine (BMAA) is a non-protein amino acid produced by cyanobacteria, diatoms and dinoflagellates. BMAA has potential to biomagnify in a terrestrial food chain, and to bioaccumulate in fish and shellfish. We have reported that administration of [ 14 C]l-BMAA to lactating mice and rats results in a mother to off-spring transfer via the milk. A preferential enantiomer-specific uptake of [ 14 C]l-BMAA has also been demonstrated in differentiated murine mammary epithelium HC11 cells. These findings, together with neurotoxic effects of BMAA demonstrated both in vitro and in vivo, highlight the need to determine whether such transfer could also occur in humans. Here, we used four cell lines of human origin to examine and compare the transport of the two BMAA enantiomers in vitro. The uptake patterns of [ 14 C]l- and [ 14 C]d-BMAA in the human mammary MCF7 cell line were in agreement with the results in murine HC11 cells, suggesting a potential secretion of BMAA into human breast milk. The permeability coefficients for both [ 14 C]l- and [ 14 C]d-BMAA over monolayers of human intestinal Caco2 cells supported an efficient absorption from the human intestine. As a final step, transport experiments confirmed that [ 14 C]l-and [ 14 C]d-BMAA can be taken up by human SHSY5Y neuroblastoma cells and even more efficiently by human U343 glioblastoma cells. In competition experiments with various amino acids, the ASCT2 specific inhibitor benzylserine was the most effective inhibitor of [ 14 C]l-BMAA uptake tested here. Altogether, our results suggest that BMAA can be transferred from an exposed mother, via the milk, to the brain of the nursed infant. Copyright © 2017. Published by Elsevier Inc.

Amino acids fortification of low-protein diet for broilers under tropical climate. 2. Nonessential amino acids and increasing essential amino acids

Directory of Open Access Journals (Sweden)

Elmutaz Atta Awad

2014-08-01

Full Text Available A three-week trial was carried out to evaluate the effect of nonessential amino acids (NEAA supplementation to a low-crude protein (CP diet with adequate essential amino acids (EAA level on growth performance, blood metabolites, and relative weights of abdominal fat, breast yield, and internal organs in broiler chickens raised under tropical hot and humid environment. Five isocaloric (3000 metabolisable energy/kg corn-soybean diets were administered (1 to 21 days to 5 groups of broilers (60 birds/group as follows: i 22.2% CP (positive control; PC; ii 16.2% CP+all EAA to meet or exceed the National Research Council (1994 recommendations (negative control; NC; iii NC+further EAA to equal the levels in the PC diet; iv NC+NEAA to equal the levels in the PC; v NC+EAA and NEAA to equal the amino acids levels in the PC diet. The results showed that the fortification of EAA alone, only improved feed intake (FI, whereas, addition of NEAA or EAA+NEAA significantly enhanced body weight, daily weight gain, and FI and decreased the feed conversion ratio to the same levels as in PC. Serum uric acid was significantly reduced and serum triglyceride increased in NC group. Dietary treatments had no significant effect on relative weights of heart, liver, abdominal fat, breast meat yield, serum albumin, and serum total protein. In conclusion, these results suggest that NEAA fortification may improve the growth performance of broilers fed an excessive low-CP diet under tropical hot and humid condition.

The Role of Amino Acids in Gluconeogenesis in Lactating Ruminants

Energy Technology Data Exchange (ETDEWEB)

Black, A. L.; Egan, A. R.; Anand, R. S.; Chapman, T. E. [Department of Physiological Sciences, School of Veterinary Medicine, University of California, Davis, CA (United States)

1968-07-01

Gluconeogenesis has an important metabolic role in all animals, but it is especially important in ruminants because of the paucity of their alimentary glucose. Several amino acids give rise to glucogenic precursors during metabolism and thus dietary or body protein represents an important source of potential glucogenic material that the ruminant can utilize to manufacture the glucose required for its physiological functions. The role of various amino acids as glucose precursors has been evaluated in lactating ruminants by making a single intravenous injection of several different amino acids uniformly labelled with {sup 14}C and following, with time, the rate and extent of incorporation of {sup 14}C into the plasma glucose. The time interval after injecting each {sup 14}C-amino acid until the specific activity maximum occurred in plasma glucose was found to vary widely among the different amino acids. Thus, the maximum specific activity in plasma glucose occurred 6 min after injection of L-aspartate-{sup 14}C and 15 min after injection of L-glutamate- {sup 14}C, while for L-valine-{sup 14}C and L-arginine-{sup 14}C the maximum specific activity in plasma glucose did not occur until 45 and 90 min, respectively, had elapsed. After injection of L-serine and L-alanine there were several maxima in the glucose specific activity. These maxima occurred between 12 and 24 min after injection of serine and during the first 30 min after injection of alanine indicating that carbon from these amino acids becomes available for glucose synthesis along diverse pathways which have different delays. Although only a few amino acids have been studied, the experimental results obtained clearly suggest an important metabolic role for protein in ruminants which has previously not been recognized. It appears that amino acids, released from protein, are utilized by the animal in a fashion which results in a prolonged availability of glucogenic precursors so that the animal can form glucose

Soft X-ray-induced decomposition of amino acids: An XPS, mass spectrometry, and NEXAFS study

International Nuclear Information System (INIS)

Zubavichus, Yan; Fuchs, Oliver; Weinhardt, Lothar; Heske, Clemens; Umbach, Eberhard; Denlinger, Jonathan D.; Grunze, Michael

2003-01-01

Decomposition of five amino acids, alanine, serine, cysteine, aspartic acid, and asparagine, under irradiation with soft X-rays (magnesium Ka X-ray source) in ultra-high vacuum was studied by means of X-ray photoelectron spectrometry (XPS) and mass spectrometry. A comparative analysis of changes in XPS line shapes, stoichiometry and residual gas composition indicates that the molecules decompose by several pathways. Dehydration, decarboxylation, decarbonylation,deamination and desulfurization of pristine molecules accompanied by desorption of H2, H2O, CO2, CO, NH3and H2S are observed with rates depending on the specific amino acid. NEXAFS spectra of cysteine at the carbon, oxygen and nitrogen K-shell and sulfur L2,3 edges complement the XPS and mass spectrometry data and show that the exposure of the sample to an intense soft X-ray synchrotron beam results in the formation of C-C and C-N double and triple bonds. Qualitatively, the amino acids studied can be arranged in the following ascending order of radiation stability:serine< alanine< aspartic acid< cysteine< asparagine

21 CFR 862.1515 - Nitrogen (amino-nitrogen) test system.

Science.gov (United States)

2010-04-01

... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Nitrogen (amino-nitrogen) test system. 862.1515... Systems § 862.1515 Nitrogen (amino-nitrogen) test system. (a) Identification. A nitrogen (amino-nitrogen) test system is a device intended to measure amino acid nitrogen levels in serum, plasma, and urine...

Serum amino acid abnormalities in pediatric patients with chronic ...

African Journals Online (AJOL)

Background: Plasma amino acid concentrations have been reported to be abnormal in patients with chronic renal failure. L-Arginine has been used to improve endothelial function by increasing nitric oxide (NO) bioavailability. The present study aim at investigating the status of plasma amino acids in pediatric patients with ...

Preliminary Studies on Some Haematological and Serum ...

African Journals Online (AJOL)

The mean serum values of sodium, potassium, chloride and bicarbonate, as well as urea, creatinine and alanine aminotransferase(ALT) enzyme did not differ from the normal values, while the mean aspartate aminotransferase (AST) enzyme value appeared to be lower than the normal range.In conclusion, the RBC, PCV ...

New Functions and Potential Applications of Amino Acids.

Science.gov (United States)

Uneyama, Hisayuki; Kobayashi, Hisamine; Tonouchi, Naoto

Currently, several types of amino acids are being produced and used worldwide. Nevertheless, several new functions of amino acids have been recently discovered that could result in other applications. For example, oral stimulation by glutamate triggers the cephalic phase response to prepare for food digestion. Further, the stomach and intestines have specific glutamate-recognizing systems in their epithelial mucosa. Regarding clinical applications, addition of monosodium glutamate to the medicinal diet has been shown to markedly enhance gastric secretion in a vagus-dependent manner. Branched-chain amino acids (BCAAs) are the major components of muscles, and ingestion of BCAAs has been found to be effective for decreasing muscle pain. BCAAs are expected to be a solution for the serious issue of aging. Further, ingestion of specific amino acids could be beneficial. Glycine can be ingested for good night's sleep: glycine ingestion before bedtime significantly improved subjective sleep quality. Ingestion of alanine and glutamine effectively accelerates alcohol metabolism, and ingestion of cystine and theanine effectively prevents colds. Finally, amino acids could be used in a novel clinical diagnostic method: the balance of amino acids in the blood could be an indicator of the risk of diseases such as cancer. These newly discovered functions of amino acids are expected to contribute to the resolution of various issues.

A gene duplication led to specialized gamma-aminobutyrate and beta-alanine aminotransferase in yeast

DEFF Research Database (Denmark)

Andersen, Gorm; Andersen, Birgit; Dobritzsch, D.

2007-01-01

and related yeasts have two different genes/enzymes to apparently 'distinguish' between the two reactions in a single cell. It is likely that upon duplication similar to 200 million years ago, a specialized Uga1p evolved into a 'novel' transaminase enzyme with broader substrate specificity.......In humans, beta-alanine (BAL) and the neurotransmitter gamma-aminobutyrate (GABA) are transaminated by a single aminotransferase enzyme. Apparently, yeast originally also had a single enzyme, but the corresponding gene was duplicated in the Saccharomyces kluyveri lineage. SkUGA1 encodes a homologue...... to characterize the substrate specificity and kinetic parameters of the four enzymes. It was found that the cofactor pyridoxal 5'-phosphate is needed for enzymatic activity and alpha-ketoglutarate, and not pyruvate, as the amino group acceptor. SkPyd4p preferentially uses BAL as the amino group donor (V...

Investigation of the adsorption of amino acids on Pd(1 1 1): A density functional theory study

International Nuclear Information System (INIS)

James, Joanna N.; Han, Jeong Woo; Sholl, David S.

2014-01-01

Graphical abstract: - Highlights: • Density functional theory calculations have been performed to study the adsorption of glycine, alanine, norvaline, valine, proline, cysteine, and serine on Pd(1 1 1). • The common structural framework for the adsorption of most amino acids onto Pd(1 1 1) is in a tridentate fashion via a nitrogen atom and two oxygen atoms. • The shifts in vibrational frequencies associated with NHH and COO support the adsorption patterns of amino acids we examined. • The adsorption strength of amino acids depends on how much the molecules deform during the adsorption process. - Abstract: Density functional theory calculations have been used to study the adsorption of glycine, alanine, norvaline, valine, proline, cysteine, and serine on Pd(1 1 1). Most amino acids except cysteine adsorb onto the surface in a tridentate fashion through a nitrogen atom and both oxygen atoms. For cysteine, an additional bond is formed with the surface due to the strong affinity of the sulfur atom, resulting in a significantly larger adsorption energy. The adsorption patterns of amino acids we examined are supported by the shifts in vibrational frequencies associated with NHH and COO. The adsorption strength of amino acids depends on how much the molecules deform during the adsorption process. Understanding the adsorption of amino acids on Pd(1 1 1) provides fundamental information for future consideration of the interactions between their derivatives or more complicated biomolecules and metal surfaces

Alanine increases blood pressure during hypotension

Science.gov (United States)

Conlay, L. A.; Maher, T. J.; Wurtman, R. J.

1990-01-01

The effect of L-alanine administration on blood pressure (BP) during haemorrhagic shock was investigated using anesthetized rats whose left carotid arteries were cannulated for BP measurement, blood removal, and drug administration. It was found that L-alanine, in doses of 10, 25, 50, 100, and 200 mg/kg, increased the systolic BP of hypotensive rats by 38 to 80 percent (while 100 mg/kg pyruvate increased BP by only 9.4 mmhg, not significantly different from saline). The results suggest that L-alanine might influence cardiovascular function.

Expression, crystallization and preliminary X-ray crystallographic analysis of Xoo0352, d-alanine-d-alanine ligase A, from Xanthomonas oryzae pv. oryzae

International Nuclear Information System (INIS)

Doan, Thanh Thi Ngoc; Kim, Jin-Kwang; Kim, Hyesoon; Ahn, Yeh-Jin; Kim, Jeong-Gu; Lee, Byoung-Moo; Kang, Lin-Woo

2008-01-01

Xoo0352, which encodes d-alanine-d-alanine ligase A (DdlA), from X. oryzae pv. oryzae was cloned, purified and crystallized. Preliminary X-ray crystallographic analysis of DdlA crystals was performed. Xanthomonas oryzae pv. oryzae (Xoo) causes bacterial blight (BB), which is one of the most devastating diseases of rice in most rice-growing countries. d-Alanine-d-alanine ligase A (DdlA), coded by the Xoo0352 gene, was expressed, purified and crystallized. DdlA is an enzyme that is involved in d-alanine metabolism and the biosynthesis of an essential bacterial peptidoglycan precursor, in which it catalyzes the formation of d-alanyl-d-alanine from two d-alanines, and is thus an attractive antibacterial drug target against Xoo. The DdlA crystals diffracted to 2.3 Å resolution and belonged to the primitive tetragonal space group P4 3 2 1 2, with unit-cell parameters a = b = 83.0, c = 97.6 Å. There is one molecule in the asymmetric unit, with a corresponding V M of 1.88 Å 3 Da −1 and a solvent content of 34.6%. The initial structure was determined by molecular replacement using d-alanine-d-alanine ligase from Staphylococcus aureus as a template model

Racemization of alanine by the alanine racemases from Salmonella typhimurium and Bacillus stearothermophilus: energetic reaction profiles

International Nuclear Information System (INIS)

Faraci, W.S.; Walsh, C.T.

1988-01-01

Alanine racemases are bacterial pyridoxal 5'-phosphate (PLP) dependent enzymes providing D-alanine as an essential building block for biosynthesis of the peptidoglycan layer of the cell wall. Two isozymic alanine racemases, encoded by the dadB gene and the alr gene, from the Gram-negative mesophilic Salmonella typhimurium and one from the Gram-positive thermophilic Bacillus stearothermophilus have been examined for the racemization mechanism. Substrate deuterium isotope effects and solvent deuterium isotope effects have been measured in both L → D and D→ L directions for all three enzymes to assess the degree to which abstraction of the α-proton or protonation of substrate PLP carbanion is limiting in catalysis. Additionally, experiments measuring internal return of α- 3 H from substrate to product and solvent exchange/substrate conversion experiments in 3 H 2 O have been used with each enzyme to examine the partitioning of substrate PLP carbanion intermediates and to obtain the relative heights of kinetically significant energy barriers in alanine racemase catalysis

Volumetric studies of some amino acids in binary aqueous solutions ...

Indian Academy of Sciences (India)

Unknown

0 values of glycine, L-alanine, and L-valine in aqueous MgCl2⋅6H2O solutions at. 298⋅15 K in order to describe the temperature dependence behaviour of partial molar quantities. Group contributions to partial molar volumes have been determined for the amino acids. The trends of transfer volumes (∆Vφ. 0) have been ...

Phosphorescence Imaging of Living Cells with Amino Acid-Functionalized Tris(2-phenylpyridine)iridium(III) Complexes

NARCIS (Netherlands)

Steunenberg, P.; Ruggi, A.; Berg, van den N.S.; Buckle, T.; Kuil, J.; Leeuwen, van F.W.B.; Velders, A.H.

2012-01-01

A series of nine luminescent cyclometalated octahedral iridium(III) tris(2-phenylpyridine) complexes has been synthesized, functionalized with three different amino acids (glycine, alanine, and lysine), on one, two, or all three of the phenylpyridine ligands. All starting complexes and final

Determination of amino acids in industrial effluents contaminated soil

International Nuclear Information System (INIS)

Mahar, M.T.; Khuhawar, M.Y.

2014-01-01

38 samples of soil for 19 locations partially irrigated on the effluents of sugar mill and oil andghee mill, bottom sediments of evaporation ponds of sugar and fertilizer industries were collected and analyzed for amino acids after acid digestion by gas chromatography using pre column derivatization with trifluroacetyleacetone and ethyl chloroformate. The results obtained were compared with the soil samples irrigated with fresh water. The soil samples were also analyzed for pH, total nitrogen contents and organic carbon. Nine essential (leucine (Leu), threonine (Thr), lysine (Lys), L-phenylalanine (Phe), tryptophan (Trp), histadine (His), L-valine (Val), methionine (Met) and isoleucine Ile) and ten non-essential ( alanine (Ala), cysteine (Cys), asparagine (Asn), glutamic acid (Glu), serine (Ser), glycine (Gly), proline (Pro), Glutamine (Gln), aspartic acid (Asp), tyrosine (Tyr)) amino acids were analyzed 13-15 amino acids were identified and determined quantitatively from soil samples. Amino acids Met, Asn, Gln and Trp were observed absent from all the samples. The variation in the amino acids contents in soil with the industrial effluents added and total nitrogen and organic carbon is discussed. (author)

Acute effect of copper exposure on serum biochemical characteristics of common carp (Cyprinus carpio L.

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Melika Ghelichpour

2014-07-01

Full Text Available Effects of ambient copper was investigated on serum stress markers, sodium and enzyme levels in Common carp (Cyprinus carpio L. over a 14-d exposure period. Fish were exposed to 0, 25 and 100 μg L-1 copper (as copper sulfate and blood was sampled at 0, 3, 7 and 14 d after exposure. Serum profile was significantly affected by copper concentration, sampling time and their interaction. Increase in serum levels of cortisol, glucose, alanine aminotransferase and aspartate aminotransferase and decrease in serum sodium levels were observed in both copper-exposed groups, 3 d after copper exposure, which lasted until the end of the experiment. It is concluded that copper exposure causes stress response and sodium loss in common carp. Likewise alanine aminotransferase and aspartate aminotransferase increase after exposure which might be as results of either tissue damage or stress.

Alanine metabolism in acute falciparum malaria

NARCIS (Netherlands)

Pukrittayakamee, S.; Krishna, S.; ter Kuile, F.; Wilaiwan, O.; Williamson, D. H.; White, N. J.

2002-01-01

We investigated the integrity of the gluconeogenic pathway in severe malaria using alanine metabolism as a measure. Alanine disposition and liver blood flow, assessed by indocyanine green (ICG) clearance, were measured simultaneously in 10 patients with falciparum malaria (six severe and four

In-Situ Measurements of the Radiation Stability of Amino Acids at 15-140 K

Science.gov (United States)

Gerakines, Perry A.; Hudson, Reggie L.; Moore, Marla H.; Bell, Jan-Luca

2012-01-01

We present new kinetics data on the radiolytic destruction of amino acids measured in situ with infrared spectroscopy. Samples were irradiated at 15, 100, and 140 K with D.8-MeV protons, and amino-acid decay was followed at each temperature with and without H2O present. Observed radiation products included CO2 and amines, consistent with amino-acid decarboxylation. The half-lives of glycine, alanine, and phenylalanine were estimated for various extraterrestrial environments. Infrared spectral changes demonstrated the conversion from the non-zwitterion structure NH2-CH2(R)-COOH at 15 K to the zwitterion structure +NH3-CH2(R)-COO- at 140 K for each amino acid studied.

Amino acid assisted dehalogenation of carbon tetrachloride by green rust

DEFF Research Database (Denmark)

Yin, Weizhao; Strobel, Bjarne W.; Hansen, Hans Chr. Bruun

2017-01-01

that reduce the formation of toxic by-products such as chloroform (CF). In this study, carbon tetrachloride (CT) dehalogenation by the chloride form of GR (GRCl) was tested in presence of glycine (GLY) and other selected amino acids. GLY, alanine (ALA) or serine (SER) all resulted in remarkable suppression...... of CF formation with only ~ 10% of CF recovery while sarcosine (SAR) showed insignificant effects. For two non-amino acid buffers, TRIS had little effect while HEPES resulted in a 40 times lower rate constant compared to experiments where no buffer was added. The FeII complexing properties of the amino...... acids and buffers caused variable extents of GRCl dissolution which was linearly correlated with CF suppression and dehalogenation rate. We hypothesize that the CF suppression seen for amino acids is caused by stabilization of carbene intermediates via the carbonyl group. Different effects on CF...

Amino and Fatty Acids of Wild Edible Mushrooms of the Genus Boletus

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Dmitri O. Levitsky

2010-10-01

Full Text Available A comparative study on the free amino acids of 15 wild edible mushroom species belonging to the genus Boletus (phylum Basidiomycota was developed. The major amino acids in the fruit bodies were arginine , alanine, glutamine, and glutamic acid. The most abundant fatty acids were oleic ( 9- 18:1, linoleic acid (9,12-18:2 , and palmitic acid (16:0, but a great variation of the ester composition from one to another one was found. Chemical constituents were characterized by GC-MS, and other chemical methods.

Topology of AspT, the aspartate:alanine antiporter of Tetragenococcus halophilus, determined by site-directed fluorescence labeling.

Science.gov (United States)

Nanatani, Kei; Fujiki, Takashi; Kanou, Kazuhiko; Takeda-Shitaka, Mayuko; Umeyama, Hideaki; Ye, Liwen; Wang, Xicheng; Nakajima, Tasuku; Uchida, Takafumi; Maloney, Peter C; Abe, Keietsu

2007-10-01

The gram-positive lactic acid bacterium Tetragenococcus halophilus catalyzes the decarboxylation of L-aspartate (Asp) with release of L-alanine (Ala) and CO(2). The decarboxylation reaction consists of two steps: electrogenic exchange of Asp for Ala catalyzed by an aspartate:alanine antiporter (AspT) and intracellular decarboxylation of the transported Asp catalyzed by an L-aspartate-beta-decarboxylase (AspD). AspT belongs to the newly classified aspartate:alanine exchanger family (transporter classification no. 2.A.81) of transporters. In this study, we were interested in the relationship between the structure and function of AspT and thus analyzed the topology by means of the substituted-cysteine accessibility method using the impermeant, fluorescent, thiol-specific probe Oregon Green 488 maleimide (OGM) and the impermeant, nonfluorescent, thiol-specific probe [2-(trimethylammonium)ethyl]methanethiosulfonate bromide. We generated 23 single-cysteine variants from a six-histidine-tagged cysteineless AspT template. A cysteine position was assigned an external location if the corresponding single-cysteine variant reacted with OGM added to intact cells, and a position was assigned an internal location if OGM labeling required cell lysis. The topology analyses revealed that AspT has a unique topology; the protein has 10 transmembrane helices (TMs), a large hydrophilic cytoplasmic loop (about 180 amino acids) between TM5 and TM6, N and C termini that face the periplasm, and a positively charged residue (arginine 76) within TM3. Moreover, the three-dimensional structure constructed by means of the full automatic modeling system indicates that the large hydrophilic cytoplasmic loop of AspT possesses a TrkA_C domain and a TrkA_C-like domain and that the three-dimensional structures of these domains are similar to each other even though their amino acid sequences show low similarity.

Alanine EPR dosimetry of therapeutic irradiators

International Nuclear Information System (INIS)

Bugay, O.; Bartchuk, V.; Kolesnik, S.; Mazin, M.; Gaponenko, H.

1999-01-01

The high-dose alanine EPR dosimetry is a very precise method in the dose range 1-100 kGy. The system is used generally as the standard high-dose transfer dosimetry in many laboratories. This is comparatively expensive technique so it is important to use it as a more universal dosimetry system also in the middle and low dose ranges. The problems of the middle-dose alanine dosimetry are discussed and the solution of several problems is proposed. The alanine EPR dosimetry has been applied to the dose measurements of medical irradiators in the Kiev City Oncology Center. (author)

Tuning electronic transport via hepta-alanine peptides junction by tryptophan doping.

Science.gov (United States)

Guo, Cunlan; Yu, Xi; Refaely-Abramson, Sivan; Sepunaru, Lior; Bendikov, Tatyana; Pecht, Israel; Kronik, Leeor; Vilan, Ayelet; Sheves, Mordechai; Cahen, David

2016-09-27

Charge migration for electron transfer via the polypeptide matrix of proteins is a key process in biological energy conversion and signaling systems. It is sensitive to the sequence of amino acids composing the protein and, therefore, offers a tool for chemical control of charge transport across biomaterial-based devices. We designed a series of linear oligoalanine peptides with a single tryptophan substitution that acts as a "dopant," introducing an energy level closer to the electrodes' Fermi level than that of the alanine homopeptide. We investigated the solid-state electron transport (ETp) across a self-assembled monolayer of these peptides between gold contacts. The single tryptophan "doping" markedly increased the conductance of the peptide chain, especially when its location in the sequence is close to the electrodes. Combining inelastic tunneling spectroscopy, UV photoelectron spectroscopy, electronic structure calculations by advanced density-functional theory, and dc current-voltage analysis, the role of tryptophan in ETp is rationalized by charge tunneling across a heterogeneous energy barrier, via electronic states of alanine and tryptophan, and by relatively efficient direct coupling of tryptophan to a Au electrode. These results reveal a controlled way of modulating the electrical properties of molecular junctions by tailor-made "building block" peptides.

Study on the EPR/dosimetric properties of some substituted alanines

International Nuclear Information System (INIS)

Gancheva, Veselka; Sagstuen, Einar; Yordanov, Nicola D.

2006-01-01

Polycrystalline phenyl-alanine and perdeuterated l-α-alanine (l-α-alanine-d 4 ) were studied as potential high-energy radiation-sensitive materials (RSM) for solid state/EPR dosimetry. It was found that phenyl-alanine exhibits a linear dose response in the dose region 0.1-17kGy. However, phenyl-alanine is about 10 times less sensitive to γ-irradiation than standard l-α-alanine irradiated at the same doses. Moreover, the EPR response from phenyl-alanine is unstable and, independent of the absorbed dose, decreases by about 50% within 20 days after irradiation upon storage at room temperature. γ-irradiated polycrystalline perdeuterated l-α-alanine (CD 3 CD(NH 2 )COOH) has not previously been studied at room temperature by EPR spectroscopy. The first part of the present analysis was with respect to the structure of the EPR spectrum. By spectrum simulations, the presence of at least two radiation induced free radicals, R 1 =CH 3 C*(H)COOH and R 2 =H 3 N + -C*(CH 3 )COO - , was confirmed very clearly. Both these radicals were suggested previously from EPR and ENDOR studies of standard alanine crystals. The further investigations into the potential use of alanine-d 4 as RSM, after choosing optimal EPR spectrometer settings parameters for this purpose, show that it is ca. two times more sensitive than standard l-α-alanine

Comparison of Free Total Amino Acid Compositions and Their Functional Classifications in 13 Wild Edible Mushrooms

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Liping Sun

2017-02-01

Full Text Available Thirteen popular wild edible mushroom species in Yunnan Province, Boletus bicolor, Boletus speciosus, Boletus sinicus, Boletus craspedius, Boletus griseus, Boletus ornatipes, Xerocomus, Suillus placidus, Boletinus pinetorus, Tricholoma terreum, Tricholomopsis lividipileata, Termitomyces microcarpus, and Amanita hemibapha, were analyzed for their free amino acid compositions by online pre-column derivazation reversed phase high-performance liquid chromatography (RP-HPLC analysis. Twenty free amino acids, aspartic acid, glutamic acid, serine, glycine, alanine, praline, cysteine, valine, methionine, phenylalanine, isoleucine, leucine, lysine, histidine, threonine, asparagines, glutamine, arginine, tyrosine, and tryptophan, were determined. The total free amino acid (TAA contents ranged from 1462.6 mg/100 g in B. craspedius to 13,106.2 mg/100 g in T. microcarpus. The different species showed distinct free amino acid profiles. The ratio of total essential amino acids (EAA to TAA was 0.13–0.41. All of the analyzed species showed high contents of hydrophobic amino acids, at 33%–54% of TAA. Alanine, cysteine, glutamine, and glutamic acid were among the most abundant amino acids present in all species. The results showed that the analyzed mushrooms possessed significant free amino acid contents, which may be important compounds contributing to the typical mushroom taste, nutritional value, and potent antioxidant properties of these wild edible mushrooms. Furthermore, the principal component analysis (PCA showed that the accumulative variance contribution rate of the first four principal components reached 94.39%. Cluster analysis revealed EAA composition and content might be an important parameter to separate the mushroom species, and T. microcarpus and A. hemibapha showed remarkable EAA content among the 13 species.

Serine and alanine racemase activities of VanT: a protein necessary for vancomycin resistance in Enterococcus gallinarum BM4174.

Science.gov (United States)

Arias, C A; Weisner, J; Blackburn, J M; Reynolds, P E

2000-07-01

Vancomycin resistance in Enterococcus gallinarum results from the production of UDP-MurNAc-pentapeptide[D-Ser]. VanT, a membrane-bound serine racemase, is one of three proteins essential for this resistance. To investigate the selectivity of racemization of L-Ser or L-Ala by VanT, a strain of Escherichia coli TKL-10 that requires D-Ala for growth at 42 degrees C was used as host for transformation experiments using plasmids containing the full-length vanT from Ent. gallinarum or the alanine racemase gene (alr) of Bacillus stearothermophilus: both plasmids were able to complement E. coli TKL-10 at 42 degrees C. No alanine or serine racemase activities were detected in the host strain E. coli TKL-10 grown at 30, 34 or 37 degrees C. Serine and alanine racemase activities were found almost exclusively (96%) in the membrane fraction of E. coli TKL-10/pCA4(vanT): the alanine racemase activity of VanT was 14% of the serine racemase activity in both E. coli TKL-10/pCA4(vanT) and E. coli XL-1 Blue/pCA4(vanT). Alanine racemase activity was present mainly (95%) in the cytoplasmic fraction of E. coli TKL-10/pJW40(alr), with a trace (1.6%) of serine racemase activity. Additionally, DNA encoding the soluble domain of VanT was cloned and expressed in E. coli M15 as a His-tagged polypeptide and purified: this polypeptide also exhibited both serine and alanine racemase activities; the latter was approximately 18% of the serine racemase activity, similar to that of the full-length, membrane-bound enzyme. N-terminal sequencing of the purified His-tagged polypeptide revealed a single amino acid sequence, indicating that the formation of heterodimers between subunits of His-tagged C-VanT and endogenous alanine racemases from E. coli was unlikely. The authors conclude that the membrane-bound serine racemase VanT also has alanine racemase activity but is able to racemize serine more efficiently than alanine, and that the cytoplasmic domain is responsible for the racemase activity.

Systematic replacement of lysine with glutamine and alanine in Escherichia coli malate synthase G: effect on crystallization

International Nuclear Information System (INIS)

Anstrom, David M.; Colip, Leslie; Moshofsky, Brian; Hatcher, Eric; Remington, S. James

2005-01-01

Alanine and glutamine mutations were made to the same 15 lysine positions on the surface of E. coli malate synthase G and the impact on crystallization observed. The results support lysine replacement for improvement of crystallization and provide insight into site selection and type of amino-acid replacement. Two proposals recommend substitution of surface lysine residues as a means to improve the quality of protein crystals. In proposal I, substitution of lysine by alanine has been suggested to improve crystallization by reducing the entropic cost of ordering flexible side chains at crystal contacts. In proposal II, substitution of lysine by residues more commonly found in crystal contacts, such as glutamine, has been proposed to improve crystallization. 15 lysine residues on the surface of Escherichia coli malate synthase G, distributed over a variety of secondary structures, were individually mutated to both alanine and glutamine. For 28 variants, detailed studies of the effect on enzymatic activity and crystallization were conducted. This has permitted direct comparison of the relative effects of the two types of mutations. While none of the variants produced crystals suitable for X-ray structural determination, small crystals were obtained in a wide variety of conditions, in support of the general approach. Glutamine substitutions were found to be more effective than alanine in producing crystals, in support of proposal II. Secondary structure at the site of mutation does not appear to play a major role in determining the rate of success

Effect of nerve growth factor on the synthesis of amino acids in PC12 cells

International Nuclear Information System (INIS)

Zielke, H.R.; Tildon, J.T.; Kauffman, F.C.; Baab, P.J.

1989-01-01

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)

Extraterrestrial Amino Acids in the Almahata Sitta Meteorite

Science.gov (United States)

Glavin, Daniel P.; Aubrey, Andrew D.; Callahan, Michael P.; Dworkin, Jason P.; Elsila, Jamie E.; Parker, Eric T.; Bada, Jeffrey L.

2010-01-01

Amino acid analysis of a meteorite fragment of asteroid 2008 TC3 called Almahata Sitta was carried out using reverse-phase liquid chromatography coupled with UV fluorescence detection and time-of-flight mass spectrometry (LC-FD/ToF-MS) as part of a sample analysis consortium. LC-FD/ToF-MS analyses of hot-water extracts from the meteorite revealed a complex distribution of two- to seven-carbon aliphatic amino acids and one- to three-carbon amines with abundances ranging from 0.5 to 149 parts-per-billion (ppb). The enantiomeric ratios of the amino acids alanine, R-amino-n-butyric acid (beta-ABA), 2-amino-2-methylbutanoic acid (isovaline), and 2-aminopentanoic acid (norvaline) in the meteorite were racemic (D/L approximately 1), indicating that these amino acids are indigenous to the meteorite and not terrestrial contaminants. Several other non-protein amino acids were also identified in the meteorite above background levels including alpha-aminoisobutyric acid (alpha-AIB), 4-amino-2- methylbutanoic acid, 4-amino-3-methylbutanoic acid, and 3-, 4-, and 5-aminopentanoic acid. The total abundances of isovaline and alpha-AIB in Almahata Sitta are 1000 times lower than the abundances of these amino acids found in the CM carbonaceous chondrite Murchison. The extremely low abundances and unusual distribution of five carbon amino acids in Almahata Sitta compared to Cl, CM, and CR carbonaceous chondrites may reflect extensive thermal alteration of amino acids on the parent asteroid by partial melting during formation or subsequent impact shock heating. It is also possible that amino acids were synthesized by catalytic reactions on the parent body after asteroid 2008 TC3 cooled to lower temperatures.

Medical reference dosimetry using EPR measurements of alanine

DEFF Research Database (Denmark)

Helt-Hansen, Jakob; Rosendal, F.; Kofoed, I.M.

2009-01-01

Background. Electron spin resonance (EPR) is used to determine the absorbed dose of alanine dosimeters exposed to clinical photon beams in a solid-water phantom. Alanine is potentially suitable for medical reference dosimetry, because of its near water equivalence over a wide energy spectrum, low...... methods the proposed algorithm can be applied without normalisation of phase shifts caused by changes in the g-value of the cavity. The study shows that alanine dosimetry is a suitable candidate for medical reference dosimetry especially for quality control applications.......Background. Electron spin resonance (EPR) is used to determine the absorbed dose of alanine dosimeters exposed to clinical photon beams in a solid-water phantom. Alanine is potentially suitable for medical reference dosimetry, because of its near water equivalence over a wide energy spectrum, low...

Amino acid substitutions in genetic variants of human serum albumin and in sequences inferred from molecular cloning

International Nuclear Information System (INIS)

Takahashi, N.; Takahashi, Y.; Blumberg, B.S.; Putnam, F.W.

1987-01-01

The structural changes in four genetic variants of human serum albumin were analyzed by tandem high-pressure liquid chromatography (HPLC) of the tryptic peptides, HPLC mapping and isoelectric focusing of the CNBr fragments, and amino acid sequence analysis of the purified peptides. Lysine-372 of normal (common) albumin A was changed to glutamic acid both in albumin Naskapi, a widespread polymorphic variant of North American Indians, and in albumin Mersin found in Eti Turks. The two variants also exhibited anomalous migration in NaDodSO 4 /PAGE, which is attributed to a conformational change. The identity of albumins Naskapi and Mersin may have originated through descent from a common mid-Asiatic founder of the two migrating ethnic groups, or it may represent identical but independent mutations of the albumin gene. In albumin Adana, from Eti Turks, the substitution site was not identified but was localized to the region from positions 447 through 548. The substitution of aspartic acid-550 by glycine was found in albumin Mexico-2 from four individuals of the Pima tribe. Although only single-point substitutions have been found in these and in certain other genetic variants of human albumin, five differences exist in the amino acid sequences inferred from cDNA sequences by workers in three other laboratories. However, our results on albumin A and on 14 different genetic variants accord with the amino acid sequence of albumin deduced from the genomic sequence. The apparent amino acid substitutions inferred from comparison of individual cDNA sequences probably reflect artifacts in cloning or in cDNA sequence analysis rather than polymorphism of the coding sections of the albumin gene

Alanine racemase mutants of Burkholderia pseudomallei and Burkholderia mallei and use of alanine racemase as a non-antibiotic-based selectable marker.

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Sheryl L W Zajdowicz

Full Text Available Burkholderia pseudomallei and Burkholderia mallei are category B select agents and must be studied under BSL3 containment in the United States. They are typically resistant to multiple antibiotics, and the antibiotics used to treat B. pseudomallei or B. mallei infections may not be used as selective agents with the corresponding Burkholderia species. Here, we investigated alanine racemase deficient mutants of B. pseudomallei and B. mallei for development of non-antibiotic-based genetic selection methods and for attenuation of virulence. The genome of B. pseudomallei K96243 has two annotated alanine racemase genes (bpsl2179 and bpss0711, and B. mallei ATCC 23344 has one (bma1575. Each of these genes encodes a functional enzyme that can complement the alanine racemase deficiency of Escherichia coli strain ALA1. Herein, we show that B. pseudomallei with in-frame deletions in both bpsl2179 and bpss0711, or B. mallei with an in-frame deletion in bma1575, requires exogenous D-alanine for growth. Introduction of bpsl2179 on a multicopy plasmid into alanine racemase deficient variants of either Burkholderia species eliminated the requirement for D-alanine. During log phase growth without D-alanine, the viable counts of alanine racemase deficient mutants of B. pseudomallei and B. mallei decreased within 2 hours by about 1000-fold and 10-fold, respectively, and no viable bacteria were present at 24 hours. We constructed several genetic tools with bpsl2179 as a selectable genetic marker, and we used them without any antibiotic selection to construct an in-frame ΔflgK mutant in the alanine racemase deficient variant of B. pseudomallei K96243. In murine peritoneal macrophages, wild type B. mallei ATCC 23344 was killed much more rapidly than wild type B. pseudomallei K96243. In addition, the alanine racemase deficient mutant of B. pseudomallei K96243 exhibited attenuation versus its isogenic parental strain with respect to growth and survival in murine

Alanine Racemase Mutants of Burkholderia pseudomallei and Burkholderia mallei and Use of Alanine Racemase as a Non-Antibiotic-Based Selectable Marker

Science.gov (United States)

Zajdowicz, Sheryl L. W.; Jones-Carson, Jessica; Vazquez-Torres, Andres; Jobling, Michael G.; Gill, Ronald E.; Holmes, Randall K.

2011-01-01

Burkholderia pseudomallei and Burkholderia mallei are category B select agents and must be studied under BSL3 containment in the United States. They are typically resistant to multiple antibiotics, and the antibiotics used to treat B. pseudomallei or B. mallei infections may not be used as selective agents with the corresponding Burkholderia species. Here, we investigated alanine racemase deficient mutants of B. pseudomallei and B. mallei for development of non-antibiotic-based genetic selection methods and for attenuation of virulence. The genome of B. pseudomallei K96243 has two annotated alanine racemase genes (bpsl2179 and bpss0711), and B. mallei ATCC 23344 has one (bma1575). Each of these genes encodes a functional enzyme that can complement the alanine racemase deficiency of Escherichia coli strain ALA1. Herein, we show that B. pseudomallei with in-frame deletions in both bpsl2179 and bpss0711, or B. mallei with an in-frame deletion in bma1575, requires exogenous d-alanine for growth. Introduction of bpsl2179 on a multicopy plasmid into alanine racemase deficient variants of either Burkholderia species eliminated the requirement for d-alanine. During log phase growth without d-alanine, the viable counts of alanine racemase deficient mutants of B. pseudomallei and B. mallei decreased within 2 hours by about 1000-fold and 10-fold, respectively, and no viable bacteria were present at 24 hours. We constructed several genetic tools with bpsl2179 as a selectable genetic marker, and we used them without any antibiotic selection to construct an in-frame ΔflgK mutant in the alanine racemase deficient variant of B. pseudomallei K96243. In murine peritoneal macrophages, wild type B. mallei ATCC 23344 was killed much more rapidly than wild type B. pseudomallei K96243. In addition, the alanine racemase deficient mutant of B. pseudomallei K96243 exhibited attenuation versus its isogenic parental strain with respect to growth and survival in murine peritoneal macrophages

Hydration of amino acids: FTIR spectra and molecular dynamics studies.

Science.gov (United States)

Panuszko, Aneta; Adamczak, Beata; Czub, Jacek; Gojło, Emilia; Stangret, Janusz

2015-11-01

The hydration of selected amino acids, alanine, glycine, proline, valine, isoleucine and phenylalanine, has been studied in aqueous solutions by means of FTIR spectra of HDO isotopically diluted in H2O. The difference spectra procedure and the chemometric method have been applied to remove the contribution of bulk water and thus to separate the spectra of solute-affected HDO. To support interpretation of obtained spectral results, molecular dynamics simulations of amino acids were performed. The structural-energetic characteristic of these solute-affected water molecules shows that, on average, water affected by amino acids forms stronger and shorter H-bonds than those in pure water. Differences in the influence of amino acids on water structure have been noticed. The effect of the hydrophobic side chain of an amino acid on the solvent interactions seems to be enhanced because of the specific cooperative coupling of water strong H-bond chain, connecting the carboxyl and amino groups, with the clathrate-like H-bond network surrounding the hydrocarbon side chain. The parameter derived from the spectral data, which corresponds to the contributions of the population of weak hydrogen bonds of water molecules which have been substituted by the stronger ones in the hydration sphere of amino acids, correlated well with the amino acid hydrophobicity indexes.

Reactivity of amino acid anions with nitrogen and oxygen atoms.

Science.gov (United States)

Wang, Zhe-Chen; Li, Ya-Ke; He, Sheng-Gui; Bierbaum, Veronica M

2018-02-14

For many decades, astronomers have searched for biological molecules, including amino acids, in the interstellar medium; this endeavor is important for investigating the hypothesis of the origin of life from space. The space environment is complex and atomic species, such as nitrogen and oxygen atoms, are widely distributed. In this work, the reactions of eight typical deprotonated amino acids (glycine, alanine, cysteine, proline, aspartic acid, histidine, tyrosine, and tryptophan) with ground state nitrogen and oxygen atoms are studied by experiment and theory. These amino acid anions do not react with nitrogen atoms. However, the reactions of these ions with oxygen atoms show an intriguing variety of ionic products and the reaction rate constants are of the order of 10 -10 cm 3 s -1 . Density functional calculations provide detailed mechanisms of the reactions, and demonstrate that spin conversion is essential for some processes. Our study provides important data and insights for understanding the kinetic and dynamic behavior of amino acids in space environments.

Amino acid compositions in heated carbonaceous chondrites and their compound-specific nitrogen isotopic ratios

Science.gov (United States)

Chan, Queenie Hoi Shan; Chikaraishi, Yoshito; Takano, Yoshinori; Ogawa, Nanako O.; Ohkouchi, Naohiko

2016-01-01

A novel method has been developed for compound-specific nitrogen isotope compositions with an achiral column which was previously shown to offer high precision for nitrogen isotopic analysis. We applied the method to determine the amino acid contents and stable nitrogen isotopic compositions of individual amino acids from the thermally metamorphosed (above 500 °C) Antarctic carbonaceous chondrites Ivuna-like (CI)1 (or CI-like) Yamato (Y) 980115 and Ornans-like (CO)3.5 Allan Hills (ALH) A77003 with the use of gas chromatography/combustion/isotope ratio mass spectrometry. ALHA77003 was deprived of amino acids due to its extended thermal alteration history. Amino acids were unambiguously identified in Y-980115, and the δ15N values of selected amino acids (glycine +144.8 ‰; α-alanine +121.2 ‰) are clearly extraterrestrial. Y-980115 has experienced an extended period of aqueous alteration as indicated by the presence of hydrous mineral phases. It has also been exposed to at least one post-hydration short-lived thermal metamorphism. Glycine and alanine were possibly produced shortly after the accretion event of the asteroid parent body during the course of an extensive aqueous alteration event and have abstained from the short-term post-aqueous alteration heating due to the heterogeneity of the parent body composition and porosity. These carbonaceous chondrite samples are good analogs that offer important insights into the target asteroid Ryugu of the Hayabusa-2 mission, which is a C-type asteroid likely composed of heterogeneous materials including hydrated and dehydrated minerals.

Topology of AspT, the Aspartate:Alanine Antiporter of Tetragenococcus halophilus, Determined by Site-Directed Fluorescence Labeling▿ †

Science.gov (United States)

Nanatani, Kei; Fujiki, Takashi; Kanou, Kazuhiko; Takeda-Shitaka, Mayuko; Umeyama, Hideaki; Ye, Liwen; Wang, Xicheng; Nakajima, Tasuku; Uchida, Takafumi; Maloney, Peter C.; Abe, Keietsu

2007-01-01

The gram-positive lactic acid bacterium Tetragenococcus halophilus catalyzes the decarboxylation of l-aspartate (Asp) with release of l-alanine (Ala) and CO2. The decarboxylation reaction consists of two steps: electrogenic exchange of Asp for Ala catalyzed by an aspartate:alanine antiporter (AspT) and intracellular decarboxylation of the transported Asp catalyzed by an l-aspartate-β-decarboxylase (AspD). AspT belongs to the newly classified aspartate:alanine exchanger family (transporter classification no. 2.A.81) of transporters. In this study, we were interested in the relationship between the structure and function of AspT and thus analyzed the topology by means of the substituted-cysteine accessibility method using the impermeant, fluorescent, thiol-specific probe Oregon Green 488 maleimide (OGM) and the impermeant, nonfluorescent, thiol-specific probe [2-(trimethylammonium)ethyl]methanethiosulfonate bromide. We generated 23 single-cysteine variants from a six-histidine-tagged cysteineless AspT template. A cysteine position was assigned an external location if the corresponding single-cysteine variant reacted with OGM added to intact cells, and a position was assigned an internal location if OGM labeling required cell lysis. The topology analyses revealed that AspT has a unique topology; the protein has 10 transmembrane helices (TMs), a large hydrophilic cytoplasmic loop (about 180 amino acids) between TM5 and TM6, N and C termini that face the periplasm, and a positively charged residue (arginine 76) within TM3. Moreover, the three-dimensional structure constructed by means of the full automatic modeling system indicates that the large hydrophilic cytoplasmic loop of AspT possesses a TrkA_C domain and a TrkA_C-like domain and that the three-dimensional structures of these domains are similar to each other even though their amino acid sequences show low similarity. PMID:17660287

On the existence of 'L-alanine cadmium bromide'.

Science.gov (United States)

Srinivasan, Bikshandarkoil R

2013-12-01

It is argued that the recently reported nonlinear optical crystal L-alanine cadmium bromide, grown by slow solvent evaporation method at room temperature [P. Ilayabarathi, J. Chandrasekaran, Spectrochim. Acta 96A (2012) 684-689] is the well-known L-alanine crystal. The isolation of L-alanine crystal is explained due to fractional crystallization. Copyright © 2013 Elsevier B.V. All rights reserved.

N-13 labeled amino acids: biodistribution, metabolism and dosimetric considerations

International Nuclear Information System (INIS)

Rosenspire, K.C.; Gelbard, A.S.

1986-01-01

With the growing interest in metabolic imaging and with the increasing number of cyclotron/PET facilities, more studies are being performed in animal and humans using short-lived positron-emitting radionuclides. Amino acids labeled either with N-13 or C-11 are one group of compounds being used to study in vivo regional organ (i.e., brain and heart) or tumor metabolism. Of the studies previously reported using C-11 or N-13 labeled amino acids (methionine, alanine, valine, glutamate, glutamine and tryptophan), imaging was restricted mainly to the organ or tissue of interest with little information obtained about the whole-bode distribution of the label. Such data are important for studying interorgan transport of amino acids and for determining accurate dosimetric measurements after intravenous injection of labeled amino acids. The goals of the authors study were to compare the distribution of several N-13 L-amino acids and N-13 ammonia in tumor-bearing mice and to determine the metabolic fate of the label in vivo. The following amino acids were enzymatically labeled using N-13 ammonia: glutamine, glutamate, methionine, α-aminobutyric acid, valine and leucine. 30 references, 2 figures, 14 tables

Fluorimetric methods for the measurement of intermediate metabolites (lactate, pyruvate, alanine, β-hydroxybutyrate, glycerol) using a COBAS FARA centrifugal analyser

OpenAIRE

Monti, L. D.; Sandoli, P. E.; Costa, S.; Phan, V. C.; Piatti, P. M.

1993-01-01

Intermediate products of the metabolism of glucose, fat and amino-acid are important in the evaluation of such metabolic disorders as diabetes mellitus, liver disease and metabolic acidosis. In the present study, methods for the measurement of intermediate metabolites (lactate, pyruvate, alanine, β-hydroxybutyrate and glycerol) have been adapted to a fast centrifugal analyzer: the COBAS FARA. Correlation coeffcients rangedfrom 0.90 to 0.99, compared to established manual spectrophotometric me...

Combined supplementation of carbohydrate, alanine, and proline is effective in maintaining blood glucose and increasing endurance performance during long-term exercise in mice.

Science.gov (United States)

Nogusa, Yoshihito; Mizugaki, Ami; Hirabayashi-Osada, Yuri; Furuta, Chie; Ohyama, Kana; Suzuki, Katsuya; Kobayashi, Hisamine

2014-01-01

Carbohydrate supplementation is extremely important during prolonged exercise because it maintains blood glucose levels during later stages of exercise. In this study, we examined whether maintaining blood glucose levels by carbohydrate supplementation could be enhanced during long-term exercise by combining this supplementation with alanine and proline, which are gluconeogenic amino acids, and whether such a combination would affect exercise endurance performance. Male C57BL/6J mice were orally administered either maltodextrin (1.25 g/kg) or maltodextrin (1.0 g/kg) with alanine (0.225 g/kg) and proline (0.025 g/kg) 15 min before running for 170 min. Combined supplementation of maltodextrin, alanine, and proline induced higher blood glucose levels than isocaloric maltodextrin alone during the late exercise phase (100-170 min). The hepatic glycogen content of mice administered maltodextrin, alanine, and proline was higher than that of mice ingesting maltodextrin alone 60 min after beginning exercise, but the glycogen content of the gastrocnemius muscle showed no difference. We conducted a treadmill running test to determine the effect of alanine and proline on endurance performance. The test showed that running time to exhaustion of mice that were supplemented with maltodextrin (2.0 g/kg) was longer than that of mice that were supplemented with water alone. Maltodextrin supplementation (1.0 g/kg) with alanine (0.9 g/kg) and proline (0.1 g/kg) further increased running time to exhaustion compared to maltodextrin alone (2.0 g/kg). These results indicate that combined supplementation of carbohydrate, alanine, and proline is effective for maintaining blood glucose and hepatic glycogen levels and increasing endurance performance during long-term exercise in mice.

Chemical mechanism of D-amino acid oxidase from Rhodotorula gracilis: pH dependence of kinetic parameters.

Science.gov (United States)

Ramón, F; Castillón, M; De La Mata, I; Acebal, C

1998-01-01

The variation of kinetic parameters of d-amino acid oxidase from Rhodotorula gracilis with pH was used to gain information about the chemical mechanism of the oxidation of D-amino acids catalysed by this flavoenzyme. d-Alanine was the substrate used. The pH dependence of Vmax and Vmax/Km for alanine as substrate showed that a group with a pK value of 6.26-7.95 (pK1) must be unprotonated and a group with a pK of 10.8-9.90 (pK2) must be protonated for activity. The lower pK value corresponded to a group on the enzyme involved in catalysis and whose protonation state was not important for binding. The higher pK value was assumed to be the amino group of the substrate. Profiles of pKi for D-aspartate as competitive inhibitor showed that binding is prevented when a group on the enzyme with a pK value of 8.4 becomes unprotonated; this basic group was not detected in Vmax/Km profiles suggesting its involvement in binding of the beta-carboxylic group of the inhibitor. PMID:9461524

Partial alanine scan of mast cell degranulating peptide (MCD): importance of the histidine- and arginine residues.

Science.gov (United States)

Buku, Angeliki; Mendlowitz, Milton; Condie, Barry A; Price, Joseph A

2004-06-01

The influence of the two histidine and two arginine residues of mast cell degranulating peptide (MCD) in activity and binding was studied by replacing these amino acids in the MCD sequence with L-alanine. Their histamine releasing activity was determined on rat peritoneal mast cells. Their binding affinity to the FcepsilonRIalpha binding subunit of the human mast cell receptor protein, was carried out using fluorescence polarization. The histamine assay showed that replacement of His13 by Ala o ccurred without loss of activity compared with the activity of MCD. Alanine substitutions for Arg7 and His8 resulted in an approximately 40 fold increase, and for Arg16 in a 14-fold increase in histamine-releasing activity of MCD. The binding affinities of the analogs were tested by competitive displacement of bound fluorescent MCD peptide from the FcepsilonRIalpha binding protein of the mast cell receptor by the Ala analogs using fluorescence polarization. The analogs Ala8 (for His) and Ala16 (for Arg) showed the same binding affinities as MCD, whereas analog Ala7 (for Arg) and analog Ala13 (for His) showed slightly better binding affinity than the parent compound. This study showed that the introduction of alanine residues in these positions resulted in MCD agonists of diverse potency. These findings will be useful in further MCD structure-activity studies.

Robotized synthesis of [3-11C]-L-alanine using reaction of asymmetric alkylation of 11CH3I nickel complex of glycine Schiff base with S-2-N-(N'-benzylpropyl)aminobenzophenone

International Nuclear Information System (INIS)

Mosevich, I.K.; Kuznetsova, O.F.; Vasil'ev, D.A.; Anichkov, A.A.; Korsakov, M.V.

1999-01-01

Synthesis of [3- 11 C]-L-alanine based on 11 CH 3 I nickel complex (1) alkylation using different solvents (tetrahydrofuran, dimethylformamide, acetonitrile, acetone) and catalysts (potassium butylate, sodium hydride) was investigated. It was shown that synthesis of amino acids labelled with 11 C based on complex (1) use permits to obtain preparations with high degree of enantiomeric enrichment. The best results (enantiomeric excess of L-alanine up to 99 %) were obtained in reaction with acetonitrile as a solvent and potassium tret-butylate as a catalyst

Dietary supplementation of Lonicera macranthoides leaf powder improves amino acid profiles in serum and longissimus thoracis muscle of growing-finishing pigs

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Cimin Long

2016-12-01

Full Text Available A 60-days feeding trial was conducted to determine the effect of dietary supplementation of Lonicera macranthoides leaf (LML on growth performance, meat quality, and free amino acid profiles in growing-finishing pigs. L. macranthoides leaves were harvested, air-dried and milled to powder. A total of 288 crossbred barrows (Large White × Landrace with an average initial BW of 54.0 ± 3.4 kg were randomly allocated to 3 treatment groups and fed either a basal diet or a diet supplemented with 0.5% or 1.0% LML powder. Results showed that diet supplemented with LML powder did not affect average daily feed intake (ADFI and feed: gain ratio (P > 0.05 in growing-finishing pigs. Moreover, diet supplemented with LML powder showed no significant effects on carcass traits (P > 0.05 including dressing percentage, backfat thickness and loin muscle area, and meat quality traits (P > 0.05 including meat color, intramuscular fat and cooking loss. However, diet supplemented with LML powder significantly improved free amino acid profiles in serum and longissimus thoracis muscle in growing-finishing pigs. Most importantly, dietary LML powder increased total free amino acids content (P < 0.001 and essential free amino acid content (P < 0.001 in longissimus thoracis muscle. These results indicate that LML has the potential to improve the nutritional value of meat through improving free amino acid profiles.

Enantioselective Collision-Activated Dissociation of Gas-Phase Tryptophan Induced by Chiral Recognition of Protonated l-Alanine Peptides

Science.gov (United States)

Fujihara, Akimasa; Matsuyama, Hiroki; Tajiri, Michiko; Wada, Yoshinao; Hayakawa, Shigeo

2017-06-01

Enantioselective dissociation in the gas phase is important for enantiomeric enrichment and chiral transmission processes in molecular clouds regarding the origin of homochirality in biomolecules. Enantioselective collision-activated dissociation (CAD) of tryptophan (Trp) and the chiral recognition ability of l-alanine peptides ( l-Ala n ; n = 2-4) were examined using a linear ion trap mass spectrometer. CAD spectra of gas-phase heterochiral H+( d-Trp)( l-Ala n ) and homochiral H+( l-Trp)( l-Ala n ) noncovalent complexes were obtained as a function of the peptide size n. The H2O-elimination product was observed in CAD spectra of both heterochiral and homochiral complexes for n = 2 and 4, and in homochiral H+( l-Trp)( l-Ala3), indicating that the proton is attached to the l-alanine peptide, and H2O loss occurs from H+( l-Ala n ) in the noncovalent complexes. H2O loss did not occur in heterochiral H+( d-Trp)( l-Ala3), where NH3 loss and (H2O + CO) loss were the primary dissociation pathways. In heterochiral H+( d-Trp)( l-Ala3), the protonation site is the amino group of d-Trp, and NH3 loss and (H2O + CO) loss occur from H+( d-Trp). l-Ala peptides recognize d-Trp through protonation of the amino group for peptide size n = 3. NH3 loss and (H2O + CO) loss from H+( d-Trp) proceeds via enantioselective CAD in gas-phase heterochiral H+( d-Trp)( l-Ala3) at room temperature, whereas l-Trp dissociation was not observed in homochiral H+( l-Trp)( l-Ala3). These results suggest that enantioselective dissociation induced by chiral recognition of l-Ala peptides through protonation could play an important role in enantiomeric enrichment and chiral transmission processes of amino acids.

Alanine aminotransferase variants conferring diverse NUE phenotypes in Arabidopsis thaliana.

Science.gov (United States)

McAllister, Chandra H; Good, Allen G

2015-01-01

Alanine aminotransferase (AlaAT, E.C. 2.6.1.2), is a pyridoxal-5'-phosphate-dependent (PLP) enzyme that catalyzes the reversible transfer of an amino group from alanine to 2-oxoglutarate to produce glutamate and pyruvate, or vice versa. It has been well documented in both greenhouse and field studies that tissue-specific over-expression of AlaAT from barley (Hordeum vulgare, HvAlaAT) results in a significant increase in plant NUE in both canola and rice. While the physical phenotypes associated with over-expression of HvAlaAT have been well characterized, the role this enzyme plays in vivo to create a more N efficient plant remains unknown. Furthermore, the importance of HvAlaAT, in contrast to other AlaAT enzyme homologues in creating this phenotype has not yet been explored. To address the role of AlaAT in NUE, AlaAT variants from diverse sources and different subcellular locations, were expressed in the wild-type Arabidopsis thaliana Col-0 background and alaat1;2 (alaat1-1;alaat2-1) knockout background in various N environments. The analysis and comparison of both the physical and physiological properties of AlaAT over-expressing transgenic plants demonstrated significant differences between plants expressing the different AlaAT enzymes under different external conditions. This analysis indicates that the over-expression of AlaAT variants other than HvAlaAT in crop plants could further increase the NUE phenotype(s) previously observed.

Relationship between serum amino-terminal propeptide of type III procollagen and changes of left ventricular function after acute myocardial infarction

DEFF Research Database (Denmark)

Poulsen, S H; Høst, N B; Jensen, S E

2000-01-01

BACKGROUND: The amino-terminal propeptide of type III procollagen (PIIINP) is a marker of type III collagen synthesis, which has previously been shown to correlate with infarct size in nonthrombolyzed myocardial infarction (MI) and to provide prognostic information after MI. METHODS AND RESULTS......: The relationship between PIIINP and changes of left ventricular (LV) function was studied in 47 consecutive patients with first acute MI and 16 control subjects. Serum PIIINP analysis was measured daily during hospitalization and on days 90, 180, and 360. LV function was assessed by echocardiography on days 1, 5......, 90, and 360. Patients with MI were stratified according to their serum PIIINP value at day 4 (group A, 5.0 microg/L). On arrival, LV function and size were comparable between groups A (n=31) and B (n=16). LV ejection fraction, initially depressed (day 1: group A, 47...

Oxidative kinetics of amino acids by peroxydisulfate: Effect of dielectric constant

International Nuclear Information System (INIS)

Khalid, Mohammad A. A.

2008-01-01

The kinetics and mechanism of oxidation of alanine, asparagines, cysteine, glutamic acid, lysine, phenylalanine and serine by peroxydisulfate ion have been studied in aqueous acidic (sulfuric acid) medium at the temperature range 60-80C. The rate shows first order dependence on peroxydisulfate concentration and zero order dependence on amino acid concentration. The rate law observed is: -d [S2O82-] /dt = Kobs [S2O82-] [amino acid]0. An autocatalytic effect has been observed in amino acids oxidation due to formation of Schiff's base between the formed aldehyde and parent amino acid. A decrease in the dielectric constant of the medium-adding acetic acid (5-15% v/v) results in a decrease in the rate in all cases studied. Reactions were carried out at different temperature (60-80C) and the thermodynamics parameters have been calculated. The logarithm of the rate constant is linearly interrelated to the square root of the ionic strength. (author)

Identification of amino acids in Securigera securidaca, a popular medicinal herb in Iranian folk medicine

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S.E. Sadat-Ebrahimi

2014-01-01

Full Text Available Securigera securidaca (L. Degen & Dorfl grows in different parts of Iran. The seeds of the species are used in Iranian folk medicine as an anti-diabetic agent. Many studies have established hypoglycemic effects of amino acids and in the present investigation, amino acids of Securigera securidaca seeds have been evaluated. The ground seeds were extracted using petroleum ether, hot ethanol and ethanol 50%, respectively. ethanol 50% extract was chromatographed over cation exchanging resin and the resulting amino acid fraction was subjected to HPLC after OPA derivatization and the amino acids were identified by comparing to standards. The results evidenced the presence of 19 amino acids in the plant extract including alanine, arginine, asparagine, aspartic acid, citrulline, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, serine, threonine, tyrosine and valine. Considering the role of some amino acids in diabetes the above amino acids could be noted as hypoglycemic agents of the plant seeds but further studies are necessary.

Alanine-ESR dosimetry for radiotherapy IAEA experience

International Nuclear Information System (INIS)

Mehta, K.; Girzikowsky, R.; )

1997-01-01

At present, the most commonly used transfer dosimeters for radiotherapy applications are TL dosemeters. They are being used for intercomparison between SSDLs (about 70) and the IAEA dosimetry laboratory. However, there are some undesirable characteristics of this dosimetry system. We have a study in progress at the IAEA to evaluate the alanine-ESR systems as an alternative to TLDs. There are several desirable qualities which make alanine an attractive dosemeter. Preliminary data suggest that the alanine-ESR dosimetry system has the potential to replace TLDs for intercomparison amongst SSDLs in the therapy-level dose regions. (Author)

Simultaneous determination of free amino acids in Pu-erh tea and their changes during fermentation.

Science.gov (United States)

Zhu, Yuchen; Luo, Yinghua; Wang, Pengpu; Zhao, Mengyao; Li, Lei; Hu, Xiaosong; Chen, Fang

2016-03-01

Pu-erh ripened tea is produced through a unique microbial fermentation process from the sun-dried leaves of large-leaf tea species (Camellia sinensis (Linn.) var. assamica (Masters) Kitamura) in Yunnan province of China. In this study, the changes of amino acid profiles during fermentation of Pu-erh tea were investigated, based on the improved HPLC-UV method with PITC pre-column derivatization for the simultaneous determination of twenty free amino acids. Results showed that aspartic acid, glutamic acid, arginine, alanine, theanine and tyrosine were the major amino acids in tea samples. Fermentation significantly influenced on the amino acid profiles. The total free amino acid contents significantly decreased during fermentation (pfermentation and then decreased gradually. The results provided the useful information for the manipulation of fermentation process according to the changes of amino acids and acrylamide contents in Pu-erh ripened tea. Copyright © 2015 Elsevier Ltd. All rights reserved.

Isolation and complete amino acid sequence of human thymopoietin and splenin

International Nuclear Information System (INIS)

Audhya, T.; Schlesinger, D.H.; Goldstein, G.

1987-01-01

Human thymopoietin and splenin were isolated from human thymus and spleen, respectively, by monitoring tissue fractionation with a bovine thymopoietin RIA cross-reactive with human thymopoietin and splenin. Bovine thymopoietin and splenin are 49-amino acid polypeptides that differ by only 2 amino acids at positions 34 and 43; the change at position 34 in the active-site region changes the receptor specificities and biological activities. The complete amino acid sequences of purified human thymopoietin and splenin were determined and shown to be 48-amino acid polypeptides differing at four positions. Ten amino acids, constant within each species for thymopoietin and splenin, differ between the human and bovine polypeptides. The pentapeptide active side of thymopoietin (residues 32-36) is constant between the human and bovine thymopoietins, but position 34 in the active site of splenin has changed from glutamic acid in bovine splenin to alanine in human splenin, accounting for the biological activity of the human but not the bovine splenin on the human T-cell line MOLT-4

Effect of different concentrations of dl-isoleucine, dl-valine, and dl-alanine on growth and sporulation in Fusarium oxysporum f. udum (Butl.) Sn. et H.

Science.gov (United States)

Prasad, M; Chaudhary, S K

1977-01-01

D1-alanine and dl-valine, when added as an extra nitrogen for fortifying the already present inorganic nitrogen source, actually acted as growth retardant for F. oxysporum f. udum (Butl.) Sn. et H. Sporulation of microconidia was indifferently affected by these two amino acids. DI-valine stimulated microconidial formation in young cultures only. In both young and old cultures the lowest concentration of dl-valine depressed macronidial sporulation. In old cultures the lowest concentration of valine stimulated chlamydospore differentiation rapidly, higher concentrations being less effective. D1-alanine, as an additional nitrogen source, depressed both macro- and microconidal sporulation. It did not even invigorate chlamydospore formation. D1-isoleucine, on the other hand, belongs to the category of growth promoters and profuse and stimulative sporulators of macro- and microconidia. This pathogen needs very specific and preferential doses of the three amino acids, if these are used as a booster in addition to the already present nitrogen source. The response, both in terms of mycelial growth and sporulation of the three spore forms, was also conditioned by the age of the culture.

[Effect of glucocorticoides on the release of amino acids in the perfused rat hindquarter (author's transl)].

Science.gov (United States)

Thienhaus, R; Tharandt, L; Zais, U; Staib, W

1975-06-01

The release of amino acids by skeletal muscle was studied in the isolated perfused rat hindquarter. Adrenalectomy depressed the formation of glutamine and alanine as well as the efflux of all other amino acids measured. Betamethasone--a synthetic glucocorticoid--caused a significant increase in the efflux of nearly all amino acids up to the level of normal controls. The release of amino acids was also increased in perfused hindquarters of diabetic rats. On the other hand, insulin exhibited a depressing effect on the release of amino acids by hindquarters of normal rats. The metabolic integrity of the muscle tissue was proved by measuring creatine phosphate, ATP, ADP and water content as well as by the significant insulin effect on glucose uptake and on [14C]leucine incorporation into muscle proteins.

A Search for Amino Acids and Nucleobases in the Martian Meteorite Roberts Massif 04262 Using Liquid Chromatography-Mass Spectrometry

Science.gov (United States)

Callahan, Michael P.; Burton, Aaron S.; Elsila, Jamie E.; Baker, Eleni M.; Smith, Karen E.; Glavin, Daniel P.; Dworkin, Jason P.

2013-01-01

The investigation into whether Mars contains signatures of past or present life is of great interest to science and society. Amino acids and nucleobases are compounds that are essential for all known life on Earth and are excellent target molecules in the search for potential Martian biomarkers or prebiotic chemistry. Martian meteorites represent the only samples from Mars that can be studied directly in the laboratory on Earth. Here, we analyzed the amino acid and nucleobase content of the shergottite Roberts Massif (RBT) 04262 using liquid chromatography-mass spectrometry. We did not detect any nucleobases above our detection limit in formic acid extracts; however, we did measure a suite of protein and nonprotein amino acids in hot-water extracts with high relative abundances of beta-alanine and gamma-amino-eta-butyric acid. The presence of only low (to absent) levels of several proteinogenic amino acids and a lack of nucleobases suggest that this meteorite fragment is fairly uncontaminated with respect to these common biological compounds. The distribution of straight-chained amine-terminal eta-omega-amino acids in RBT 04262 resembled those previously measured in thermally altered carbonaceous meteorites. A carbon isotope ratio of -24(0/00) +/- 6(0/00) for beta-alanine in RBT 04262 is in the range of reduced organic carbon previously measured in Martian meteorites (Steele et al. 2012). The presence of eta-omega-amino acids may be due to a high temperature Fischer-Tropschtype synthesis during igneous processing on Mars or impact ejection of the meteorites from Mars, but more experimental data are needed to support these hypotheses.

Effect of Arsenic and Chromium on the Serum Amino-Transferases Activity in Indian Major Carp, Labeo rohita

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Anjaneyulu Yerramilli

2007-09-01

Full Text Available Arsenic and hexavalent chromium toxicity results from their ability to interact with sulfahydryl groups of proteins and enzymes, and to substitute phosphorus in a variety of biochemical reactions. Alanine aminotransferase (ALT; E.C: 2.6.1.2 and Aspartate amino transferase (AST; EC 2.6.1.1 play a crucial role in transamination reactions and can be used as potential biomarkers to indicate hepatotoxicity and cellular damage. While histopathological studies in liver tissue require more time and expertise, simple and reliable biochemical analysis of ALT and AST can be used for a rapid assessment of tissue and cellular damage within 96 h. The main objective of this study was to determine the acute effects of arsenic and hexavalent chromium on the activity of ALT and AST in the Indian major carp, Labeo rohita for 24 h and 96 h. Significant increase in the activity of ALT (P < 0.01 from controls in arsenic exposed fish indicates serious hepatic damage and distress condition to the fish. However, no such significant changes were observed in chromium-exposed fish suggesting that arsenic is more toxic to the fish. These findings indicate that ALT and AST are candidate biomarkers for arsenic-induced hepatotoxicity in Labeo rohita.

Corrosion control of vanadium in aqueous solutions by amino acids

International Nuclear Information System (INIS)

El-Rabiee, M.M.; Helal, N.H.; El-Hafez, Gh.M. Abd; Badawy, W.A.

2008-01-01

The electrochemical behavior of vanadium in amino acid free and amino acid containing aqueous solutions of different pH was studied using open-circuit potential measurements, polarization techniques and electrochemical impedance spectroscopy (EIS). The corrosion current density, i corr , the corrosion potential, E corr and the corrosion resistance, R corr , were calculated. A group of amino acids, namely, glycine, alanine, valine, histidine, glutamic and cysteine has been investigated as environmentally safe inhibitors. The effect of Cl - on the corrosion inhibition efficiency especially in acid solutions was investigated. In neutral and basic solutions, the presence of amino acids increases the corrosion resistance of the metal. The electrochemical behavior of V before and after the corrosion inhibition process has shown that some amino acids like glutamic acid and histidine have promising corrosion inhibition efficiency at low concentration (≅25 mM). The inhibition efficiency (η) was found to depend on the structure of the amino acid and the constituents of the corrosive medium. The corrosion inhibition process is based on the adsorption of the amino acid molecules on the metal surface and the adsorption process follows the Freundlich isotherm. The adsorption free energy for valine on V in acidic solutions was found to be -9.4 kJ/mol which reveals strong physical adsorption of the amino acid molecules on the vanadium surface

Pyridyl-alanine as a Hydrophilic, Aromatic Element in Peptide Structural Optimization.

Science.gov (United States)

Mroz, Piotr A; Perez-Tilve, Diego; Liu, Fa; Gelfanov, Vasily; DiMarchi, Richard D; Mayer, John P

2016-09-08

Glucagon (Gcg) 1 serves a seminal physiological role in buffering against hypoglycemia, but its poor biophysical properties severely complicate its medicinal use. We report a series of novel glucagon analogues of enhanced aqueous solubility and stability at neutral pH, anchored by Gcg[Aib16]. Incorporation of 3- and 4-pyridyl-alanine (3-Pal and 4-Pal) enhanced aqueous solubility of glucagon while maintaining biological properties. Relative to native hormone, analogue 9 (Gcg[3-Pal6,10,13, Aib16]) demonstrated superior biophysical character, better suitability for medicinal purposes, and comparable pharmacology against insulin-induced hypoglycemia in rats and pigs. Our data indicate that Pal is a versatile surrogate to natural aromatic amino acids and can be employed as an alternative or supplement with isoelectric adjustment to refine the biophysical character of peptide drug candidates.

The Cyanobacteria Derived Toxin Beta-N-Methylamino-L-Alanine and Amyotrophic Lateral Sclerosis

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Elijah W. Stommel

2010-12-01

Full Text Available There is mounting evidence to suggest that environmental factors play a major role in the development of neurodegenerative diseases like ALS (Amyotrophic Lateral Sclerosis. The non-protein amino acid beta-N-methylamino-L-alanine (BMAA was first associated with the high incidence of Amyotrophic Lateral Sclerosis/Parkinsonism Dementia Complex (ALS/PDC in Guam, and has been implicated as a potential environmental factor in ALS, Alzheimer’s disease, and other neurodegenerative diseases. BMAA has a number of toxic effects on motor neurons including direct agonist action on NMDA and AMPA receptors, induction of oxidative stress, and depletion of glutathione. As a non-protein amino acid, there is also the strong possibility that BMAA could cause intraneuronal protein misfolding, the hallmark of neurodegeneration. While an animal model for BMAA-induced ALS is lacking, there is substantial evidence to support a link between this toxin and ALS. The ramifications of discovering an environmental trigger for ALS are enormous. In this article, we discuss the history, ecology, pharmacology and clinical ramifications of this ubiquitous, cyanobacteria-derived toxin.

Metabolism of leucine and alanine in growing rats fed the diets with various protein to energy ratios

International Nuclear Information System (INIS)

Tanaka, Hideyuki; Yamaguchi, Michio; Kametaka, Masao

1975-01-01

In order to clarify the nutritional significance of metabolism of the carbon skeleton of individual amino acids, the metabolic fates of L-leucine-U- 14 C and L-alanine-U- 14 C were investigated in growing rats fed the diets with various protein calories percents (PC%) at 410 kcal of metabolizable energy. The incorporation of 14 C into body protein in 12 hr after the injection of leucine- 14 C was about 73% of the dose in the 0 and 5 PC% groups, though it decreased with increasing the levels of dietary protein from 10 to 30 PC%. The value of 14 C recovery in body protein almost agreed with the net protein utilization (NPU) determined for the whole egg protein in a similar experimental condition. The 14 C recovery in expired CO 2 and body lipid suggested that the carbon skeleton of leucine is well utilized as an energy source when the dietary carbohydrate is extensively replaced by protein. While, the incorporation of 14 C into body protein from alanine- 14 C was less than about 11% of the dose in all the dietary groups, and the majority of 14 C was recovered in expired CO 2 and body lipid in a remarked contrast to leucine. A similar pattern in urinary excretion of 14 C was obtained for these amino acids, and the refracted rise of 14 C from 10 PC% may give an indication for minimum protein requirements. (auth.)

EFFECT OF THIOPROPANOL ON AMINO ACID TURNOVER AND REDOX STATUS IN ALLOXAN DIABETIC RAT LIVER

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Vickram

2016-07-01

Full Text Available BACKGROUND Decreased cellular thiol levels seen in diabetes mellitus (DM may be in part attributed to increased free radical generation. The free radical mediated oxidative stress has been implicated in the pathogenesis of DM and its complications. The relative deficiency or non-availability of insulin in DM affects the metabolism of biomolecules, specifically the carbohydrate metabolism. The insulin-mimicking actions of various thiols have been studied. In our previous study, we have documented that 3-mercapto- 1-propanol (Thiopropanol, a low molecular weight thiol, at the dosage employed has increased glucose utilisation in alloxandiabetic rat liver tissue probably by favouring utilisation of glucose through glycolysis and HMP pathway. It is known that insulin inhibits gluconeogenesis by inhibiting the key enzymes of the same and by controlling the channelling of amino acids for the glucose biosynthesis through gluconeogenic pathway. A study was undertaken to assess the effects of thiopropanol (TP on amino acid turnover and the redox status in alloxan diabetic rat liver. METHODS Male albino rats weighing 150-250 g were used. Diabetes was induced using alloxan monohydrate. Rats were divided into normal and diabetic groups. Levels of amino acid nitrogen (AAN, alanine, total thiol (-SH groups, TBARS (Thiobarbituric acid reactive substances, and activities of alanine transaminase (ALT and aspartate transaminase (AST were estimated in liver specimens of normal, control-alloxan diabetic and TP-exposed-alloxan-diabetic rats. RESULTS The results showed a significant increase (p<0.001 in AAN levels, alanine levels, and total -SH groups concentration; and a significant decrease (p<0.001 in TBARS levels, ALT and AST activities in TP-exposed-alloxan diabetic liver slices as compared to control-alloxan diabetic liver slices. CONCLUSIONS Hence, it may be concluded that TP, at the concentration employed, inhibits gluconeogenesis from amino acids probably by

Performance effects of acute β-alanine induced paresthesia in competitive cyclists.

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Bellinger, Phillip M; Minahan, Clare L

2016-01-01

β-alanine is a common ingredient in supplements consumed by athletes. Indeed, athletes may believe that the β-alanine induced paresthesia, experienced shortly after ingestion, is associated with its ergogenic effect despite no scientific mechanism supporting this notion. The present study examined changes in cycling performance under conditions of β-alanine induced paresthesia. Eight competitive cyclists (VO2max = 61.8 ± 4.2 mL·kg·min(-1)) performed three practices, one baseline and four experimental trials. The experimental trials comprised a 1-km cycling time trial under four conditions with varying information (i.e., athlete informed β-alanine or placebo) and supplement content (athlete received β-alanine or placebo) delivered to the cyclist: informed β-alanine/received β-alanine, informed placebo/received β-alanine, informed β-alanine/received placebo and informed placebo/received placebo. Questionnaires were undertaken exploring the cyclists' experience of the effects of the experimental conditions. A possibly likely increase in mean power was associated with conditions in which β-alanine was administered (±95% CL: 2.2% ± 4.0%), but these results were inconclusive for performance enhancement (p = 0.32, effect size = 0.18, smallest worthwhile change = 56% beneficial). A possibly harmful effect was observed when cyclists were correctly informed that they had ingested a placebo (-1.0% ± 1.9%). Questionnaire data suggested that β-alanine ingestion resulted in evident sensory side effects and six cyclists reported placebo effects. Acute ingestion of β-alanine is not associated with improved 1-km TT performance in competitive cyclists. These findings are in contrast to the athlete's "belief" as cyclists reported improved energy and the ability to sustain a higher power output under conditions of β-alanine induced paresthesia.

Effect of glycine, DL-alanine and DL-2-aminobutyric acid on the temperature of maximum density of water

International Nuclear Information System (INIS)

Romero, Carmen M.; Torres, Andres Felipe

2015-01-01

Highlights: • Effect of α-amino acids on the temperature of maximum density of water is presented. • The addition of α-amino acids decreases the temperature of maximum density of water. • Despretz constants suggest that the amino acids behave as water structure breakers. • Despretz constants decrease as the number of CH 2 groups of the amino acid increase. • Solute disrupting effect becomes smaller as its hydrophobic character increases. - Abstract: The effect of glycine, DL-alanine and DL-2-aminobutyric acid on the temperature of maximum density of water was determined from density measurements using a magnetic float densimeter. Densities of aqueous solutions were measured within the temperature range from T = (275.65 to 278.65) K at intervals of T = 0.50 K over the concentration range between (0.0300 and 0.1000) mol · kg −1 . A linear relationship between density and concentration was obtained for all the systems in the temperature range considered. The temperature of maximum density was determined from the experimental results. The effect of the three amino acids is to decrease the temperature of maximum density of water and the decrease is proportional to molality according to Despretz equation. The effect of the amino acids on the temperature of maximum density decreases as the number of methylene groups of the alkyl chain becomes larger. The results are discussed in terms of (solute + water) interactions and the effect of amino acids on water structure

Enzymatic properties of the glycine D-alanine [corrected] aminopeptidase of Aspergillus oryzae and its activity profiles in liquid-cultured mycelia and solid-state rice culture (rice koji).

Science.gov (United States)

Marui, Junichiro; Matsushita-Morita, Mayumi; Tada, Sawaki; Hattori, Ryota; Suzuki, Satoshi; Amano, Hitoshi; Ishida, Hiroki; Yamagata, Youhei; Takeuchi, Michio; Kusumoto, Ken-Ichi

2012-01-01

The gdaA gene encoding S12 family glycine-D-alanine aminopeptidase (GdaA) was found in the industrial fungus Aspergillus oryzae. GdaA shares 43% amino acid sequence identity with the D-aminopeptidase of the Gram-negative bacterium Ochrobactrum anthropi. GdaA purified from an A. oryzae gdaA-overexpressing strain exhibited high D-stereospecificity and efficiently released N-terminal glycine and D-alanine of substrates in a highly specific manner. The optimum pH and temperature were 8 to 9 and 40°C, respectively. This enzyme was stable under alkaline conditions at pH 8 to 11 and relatively resistant to acidic conditions until pH 5.0. The chelating reagent EDTA, serine protease inhibitors such as AEBSF, benzamidine, TPCK, and TLCK, and the thiol enzyme inhibitor PCMB inhibited the enzyme. The aminopeptidase inhibitor bestatin did not affect the activity. GdaA was largely responsible for intracellular glycine and D-alanine aminopeptidase activities in A. oryzae during stationary-phase growth in liquid media. In addition, the activity increased in response to the depletion of nitrogen or carbon sources in the growth media, although the GdaA-independent glycine aminopeptidase activity highly increased simultaneously. Aminopeptidases of A. oryzae attract attention because the enzymatic release of a variety of amino acids and peptides is important for the enhancement of the palatability of fermented foods. GdaA activity was found in extracts of a solid-state rice culture of A. oryzae (rice koji), which is widely used as a starter culture for Japanese traditional fermented foods, and was largely responsible for the glycine and D-alanine aminopeptidase activity detected at a pH range of 6 to 9.

NMR chemical shifts in amino acids: Effects of environments, electric field, and amine group rotation

International Nuclear Information System (INIS)

Yoon, Young-Gui; Pfrommer, Bernd G.; Louie, Steven G.; Canning, Andrew

2002-01-01

The authors present calculations of NMR chemical shifts in crystalline phases of some representative amino acids such as glycine, alanine, and alanyl-alanine. To get an insight on how different environments affect the chemical shifts, they study the transition from the crystalline phase to completely isolated molecules of glycine. In the crystalline limit, the shifts are dominated by intermolecular hydrogen-bonds. In the molecular limit, however, dipole electric field effects dominate the behavior of the chemical shifts. They show that it is necessary to average the chemical shifts in glycine over geometries. Tensor components are analyzed to get the angle dependent proton chemical shifts, which is a more refined characterization method

Occurrence of β-N-methylamino-l-alanine (BMAA and Isomers in Aquatic Environments and Aquatic Food Sources for Humans

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Emilie Lance

2018-02-01

Full Text Available The neurotoxin β-N-methylamino-l-alanine (BMAA, a non-protein amino acid produced by terrestrial and aquatic cyanobacteria and by micro-algae, has been suggested to play a role as an environmental factor in the neurodegenerative disease Amyotrophic Lateral Sclerosis-Parkinsonism-Dementia complex (ALS-PDC. The ubiquitous presence of BMAA in aquatic environments and organisms along the food chain potentially makes it public health concerns. However, the BMAA-associated human health risk remains difficult to rigorously assess due to analytical challenges associated with the detection and quantification of BMAA and its natural isomers, 2,4-diamino butyric acid (DAB, β-amino-N-methyl-alanine (BAMA and N-(2-aminoethyl glycine (AEG. This systematic review, reporting the current knowledge on the presence of BMAA and isomers in aquatic environments and human food sources, was based on a selection and a score numbering of the scientific literature according to various qualitative and quantitative criteria concerning the chemical analytical methods used. Results from the best-graded studies show that marine bivalves are to date the matrix containing the higher amount of BMAA, far more than most fish muscles, but with an exception for shark cartilage. This review discusses the available data in terms of their use for human health risk assessment and identifies knowledge gaps requiring further investigations.

Occurrence of β-N-methylamino-l-alanine (BMAA) and Isomers in Aquatic Environments and Aquatic Food Sources for Humans.

Science.gov (United States)

Lance, Emilie; Arnich, Nathalie; Maignien, Thomas; Biré, Ronel

2018-02-14

The neurotoxin β- N -methylamino-l-alanine (BMAA), a non-protein amino acid produced by terrestrial and aquatic cyanobacteria and by micro-algae, has been suggested to play a role as an environmental factor in the neurodegenerative disease Amyotrophic Lateral Sclerosis-Parkinsonism-Dementia complex (ALS-PDC). The ubiquitous presence of BMAA in aquatic environments and organisms along the food chain potentially makes it public health concerns. However, the BMAA-associated human health risk remains difficult to rigorously assess due to analytical challenges associated with the detection and quantification of BMAA and its natural isomers, 2,4-diamino butyric acid (DAB), β-amino- N -methyl-alanine (BAMA) and N -(2-aminoethyl) glycine (AEG). This systematic review, reporting the current knowledge on the presence of BMAA and isomers in aquatic environments and human food sources, was based on a selection and a score numbering of the scientific literature according to various qualitative and quantitative criteria concerning the chemical analytical methods used. Results from the best-graded studies show that marine bivalves are to date the matrix containing the higher amount of BMAA, far more than most fish muscles, but with an exception for shark cartilage. This review discusses the available data in terms of their use for human health risk assessment and identifies knowledge gaps requiring further investigations.

Alanine - Valine dynamics in pregnant rabbits | Emudianughe ...

African Journals Online (AJOL)

[15N]-alanine and [15N]–valine dynamics were studied in 29 -30 days pregnant New-Zealand rabbits. Over the experimental period, there was no detectable significant difference of mean ± SD of alanine concentrations within the sampling intervals in maternal, umbilical venous and arterial blood samples suggesting that ...

Free amino acid pools in muscle and hemolymph during the molt cycle of the land crab, Gecarcinus lateralis

Energy Technology Data Exchange (ETDEWEB)

Yamaoka, L.H.; Skinner, D.M.

1976-01-01

The total free amino acids in muscle water of the land crab, Gecarcinus lateralis, decrease almost 3-fold during the premolt period in comparison to the intermolt period (193 ..mu..M/g during intermolt and 67 ..mu..M/g during late premolt). This decrease is accounted for primarily by changes in the nonessential amino acids proline, glycine and alanine. At the same stage, several essential amino acids (lysine, methionine and tyrosine) increase 2- to 4-fold. Although free amino acid levels in hemolymph are lower and more variable than those in muscle, the same amino acids show similar molt-stage-related changes. The decreases in glycine and proline may be associated with the synthesis of the new exoskeleton and connective tissue during the premolt period.

Role of urea in controlling radiation-induced changes in some amino acids level and protein end products in blood serum of rats

International Nuclear Information System (INIS)

El-Kashef, H.S.; Saada, H.N.

1991-01-01

In previous studies (Helen Saada, 1982), urea could successfully act as a good radio-protector. It helps the recovery from changes induced by gamma irradiation in soluble protein contents, and percentage of protein fractions as well as some enzymatic activity. In the present study, it has been found that urea acts also as adequate radioprotector against radiation induced changes in the level of free and tied amino acids, glucosamine,free ammonia and urea concentration in the blood serum of white rats. The radioprotective role of urea was very clear on the first day after irradiation as manifested by the levels of free amino acids and protein end products (urea and ammonia). While in the case of tied amino acids, the action of urea as a radioprotector could be noticed lately, on the third post-irradiation day. The recovery from radiation hazards depended not only on the time lapse after irradiation but also on the chemical structure of the molecule which presents or constitutes the biological matter.2 tab.,1 fig

Amino acid metabolism during exercise in trained rats: the potential role of carnitine in the metabolic fate of branched-chain amino acids.

Science.gov (United States)

Ji, L L; Miller, R H; Nagle, F J; Lardy, H A; Stratman, F W

1987-08-01

The influence of endurance training and an acute bout of exercise on plasma concentrations of free amino acids and the intermediates of branched-chain amino acid (BCAA) metabolism were investigated in the rat. Training did not affect the plasma amino acid levels in the resting state. Plasma concentrations of alanine (Ala), aspartic acid (Asp), asparagine (Asn), arginine (Arg), histidine (His), isoleucine (Ile), leucine (Leu), lysine (Lys), methionine (Met), phenylalanine (Phe), proline (Pro), serine (Ser), threonine (Thr), and valine (Val) were significantly lower, whereas glutamate (Glu), glycine (Gly), ornithine (Orn), tryptophan (Trp), tyrosine (Tyr), creatinine, urea, and ammonia levels were unchanged, after one hour of treadmill running in the trained rats. Plasma concentration of glutamine (Glu), the branched-chain keto acids (BCKA) and short-chain acyl carnitines were elevated with exercise. Ratios of plasma BCAA/BCKA were dramatically lowered by exercise in the trained rats. A decrease in plasma-free carnitine levels was also observed. These data suggest that amino acid metabolism is enhanced by exercise even in the trained state. BCAA may only be partially metabolized within muscle and some of their carbon skeletons are released into the circulation in forms of BCKA and short-chain acyl carnitines.

Free amino acid profile of Bubalus bubalis L. meat from the Campania region

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Nicola Landi

Full Text Available ABSTRACT In this study, we determined the amount of carnosine and anserine in water buffalo meat without hanging treatment and the free amino acid profile by using amino acid analyser with post-column ninhydrin derivatization procedure. The main free amino acids present in samples were glutamic acid (>60 mg/100 g, followed by alanine, glycine, and arginine. Other protein amino acids were detected in minor amounts (less than 2 mg/100 g. Among the non-protein amine-containing compounds, taurine and urea were the most abundant. The analysis showed that 50% of the total free amino acids was represented by dipeptides carnosine (average ~130.3 mg/100 g and anserine (average ~17.9 mg/100 g. Thus, this study for the first time reports the free amino acids profile of water buffalo meat and the content of carnosine and anserine, potentially involved in the darkening meat process and their ratio, that could be used to estimate the water buffalo meat portion in mixed meat products.

IAEA reference dosimeter: Alanine-ESR

International Nuclear Information System (INIS)

Mehta, K.; Girzikowsky, R.

1999-01-01

Since 1985, the IAEA has been using alanine-ESR as a transfer dosimeter for its dose quality audit service, namely the International Dose Assurance Service. The alanine dosimeters are rod-type containing 70 wt% DL--α-alanine and 30 wt% polystyrene. We have two self-shielded gamma facilities for the calibration of the dosimetry system, where the temperature within the irradiation chamber can be controlled by a specially designed unit. A 4th order polynomial is fitted to the 16 data points in the dose range of 100 Gy to 50 kGy. The measured value of the irradiation temperature coefficient at two dose values (15 and 45 kGy) is 0.23 %/deg. C. Also, the ESR-response was followed for several dosimeters for about 8 months to study the post-irradiation effect. A value of 0.008 %/day was observed for the fading of the response for two dose values (15 and 45 kGy) and three irradiation temperatures (15, 27 and 40 deg. C). The effect of the analysis temperature on the ESR response was also studied. The combined relative uncertainty for the IAEA alanine-ESR dosimetry system is 1.5% (k=1). This includes that transferred from the primary laboratory for the dose rate measurements of the gamma facilities, dosimetry system calibration uncertainties, batch variability and uncertainty in the curve fitting procedure. This value however does not include the contribution due to the irradiation temperature correction which is applied when it differs from that during calibration; this component being specific for each dose measurement. (author)

Alterations in protein and amino acid metabolism in rats fed a branched-chain amino acid- or leucine-enriched diet during postprandial and postabsorptive states.

Science.gov (United States)

Holecek, Milan; Siman, Pavel; Vodenicarovova, Melita; Kandar, Roman

2016-01-01

Many people believe in favourable effects of branched-chain amino acids (BCAAs; valine, leucine, and isoleucine), especially leucine, on muscle protein balance and consume BCAAs for many years. We determined the effects of the chronic intake of a BCAA- or leucine-enriched diet on protein and amino acid metabolism in fed and postabsorptive states. Rats were fed a standard diet, a diet with a high content of valine, leucine, and isoleucine (HVLID), or a high content of leucine (HLD) for 2 months. Half of the animals in each group were sacrificed in the fed state on the last day, and the other half were sacrificed after overnight fast. Protein synthesis was assessed using the flooding dose method (L-[3,4,5-(3)H]phenylalanine), proteolysis on the basis of chymotrypsin-like activity (CHTLA) of proteasome and cathepsin B and L activities. Chronic intake of HVLID or HLD enhanced plasma levels of urea, alanine and glutamine. HVLID also increased levels of all three BCAA and branched-chain keto acids (BCKA), HLD increased leucine, ketoisocaproate and alanine aminotransferase and decreased valine, ketovaline, isoleucine, ketoisoleucine, and LDL cholesterol. Tissue weight and protein content were lower in extensor digitorum longus muscles in the HLD group and higher in kidneys in the HVLID and HLD groups. Muscle protein synthesis in postprandial state was higher in the HVLID group, and CHTLA was lower in muscles of the HVLID and HLD groups compared to controls. Overnight starvation enhanced alanine aminotransferase activity in muscles, and decreased protein synthesis in gastrocnemius (in HVLID group) and extensor digitorum longus (in HLD group) muscles more than in controls. Effect of HVLID and HLD on CHTLA in muscles in postabsorptive state was insignificant. The results failed to demonstrate positive effects of the chronic consumption of a BCAA-enriched diet on protein balance in skeletal muscle and indicate rather negative effects from a leucine-enriched diet. The primary

Phospholyl(borane) Amino Acids and Peptides: Stereoselective Synthesis and Fluorescent Properties with Large Stokes Shift.

Science.gov (United States)

Arribat, Mathieu; Rémond, Emmanuelle; Clément, Sébastien; Lee, Arie Van Der; Cavelier, Florine

2018-01-24

The synthesis of phospholyl(borane) amino acids was stereoselectively achieved by reaction of phospholide anion with iodo α-amino ester derived from l-aspartic acid or l-serine, followed by in situ complexation with borane. Phospholyl(borane) amino acids are easy to store and can be subjected to direct transformation into the corresponding free phospholyl, gold complex, oxide or sulfur derivatives as well as phospholinium salts, thus offering a variety of side chains. After selective deprotection of carboxylic function or amine, C- or N- peptide coupling with an alanine moiety proved the possible incorporation into peptides. Such phospholyl amino acid and peptide derivatives exhibit fluorescent properties with a large Stokes shift (160 nm) and fluorescence up to 535 nm, depending on the phosphole aromaticity and the chemical environment. These phospholyl(borane) amino acids constitute a new class of unnatural amino acids useful for structure-activities relationship studies and appear to be promising fluorophores for the development of labeled peptides.

Refractive properties of imidazolium ionic liquids with alanine anion [C n mim][Ala] ( n = 2, 3, 4, 5, 6)

Science.gov (United States)

Zhou, Cai-bin; Li, Jing; Yi, Zhe; Ai, Hong-jun

2017-10-01

Imidazolium ionic liquids with alanine anion, [C n mim][Ala] ( n = 2, 3, 4, 5, 6), were prepared and characterized. The standard addition method was used to measure refractivity of the synthesized ionic liquids in order to eliminate the effect of water traces in the ionic liquids. The molar refraction and molecular polarizability were also calculated by Lorentz-Lorenz's method. Using liner extrapolation, the semiempirical estimation method for optical properties of other ionic liquids with amino acid anions was established.

Isotopic variants of light and heavy L-pyroglutamic acid succinimidyl esters as the derivatization reagents for DL-amino acid chiral metabolomics identification by liquid chromatography and electrospray ionization mass spectrometry

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Mochizuki, Toshiki; Todoroki, Kenichiro; Inoue, Koichi; Min, Jun Zhe; Toyo’oka, Toshimasa, E-mail: toyooka@u-shizuoka-ken.ac.jp

2014-02-06

Graphical abstract: -- Highlights: •Isotopic variants of chiral labeling reagents were newly synthesized. •Analysis of DL-amino acids was performed by UPLC–ESI–MS/MS. •Highly efficient enantioseparation and detection of DL-amino acids were performed. •Differential analysis of DL-amino acid was successfully performed in real samples. -- Abstract: L-Pyroglutamic acid succinimidyl ester (L-PGA-OSu) and its isotopic variant (L-PGA[d{sub 5}]-OSu) were newly synthesized and evaluated as the chiral labeling reagents for the enantioseparation of amino acids, in terms of separation efficiency by reversed-phase chromatography and detection sensitivity by ESI-MS/MS. The enantiomers of amino acids were easily labeled with the reagents at 60 °C within 10 min in an alkaline medium containing triethylamine. Although all the diastereomers derived from 18 proteolytic amino acids could not be satisfactorily separated, the pairs of 9 amino acids were completely separated by reversed-phase chromatography using the small particle (1.7 μm) ODS column (Rs = 1.95–8.05). The characteristic daughter ions, i.e., m/z 84.04 and m/z 89.04, were detected from all the derivatives by the collision induced dissociation of the protonated molecular ions. A highly sensitive detection at a low-fmol level (0.5–3.2 fmol) was also obtained from the selected reaction monitoring (SRM) chromatograms. An isotope labeling strategy using light and heavy L-PGA-OSu for the differential analysis of the DL-amino acids in different sample groups is also presented in this paper. The differential analysis of biological sample (i.e., human serum) and food product (i.e., yogurt) were tried to demonstrate the efficiency of the proposed method. The ratios of the DL-amino acids in human serum samples, spiked with the different concentrations of D-amino acids, were determined by the procedures using L-PGA-OSu and L-PGA[d{sub 5}]-OSu. The D/L ratios in the two sample groups at different concentrations of

Isotopic variants of light and heavy L-pyroglutamic acid succinimidyl esters as the derivatization reagents for DL-amino acid chiral metabolomics identification by liquid chromatography and electrospray ionization mass spectrometry

International Nuclear Information System (INIS)

Mochizuki, Toshiki; Todoroki, Kenichiro; Inoue, Koichi; Min, Jun Zhe; Toyo’oka, Toshimasa

2014-01-01

Graphical abstract: -- Highlights: •Isotopic variants of chiral labeling reagents were newly synthesized. •Analysis of DL-amino acids was performed by UPLC–ESI–MS/MS. •Highly efficient enantioseparation and detection of DL-amino acids were performed. •Differential analysis of DL-amino acid was successfully performed in real samples. -- Abstract: L-Pyroglutamic acid succinimidyl ester (L-PGA-OSu) and its isotopic variant (L-PGA[d 5 ]-OSu) were newly synthesized and evaluated as the chiral labeling reagents for the enantioseparation of amino acids, in terms of separation efficiency by reversed-phase chromatography and detection sensitivity by ESI-MS/MS. The enantiomers of amino acids were easily labeled with the reagents at 60 °C within 10 min in an alkaline medium containing triethylamine. Although all the diastereomers derived from 18 proteolytic amino acids could not be satisfactorily separated, the pairs of 9 amino acids were completely separated by reversed-phase chromatography using the small particle (1.7 μm) ODS column (Rs = 1.95–8.05). The characteristic daughter ions, i.e., m/z 84.04 and m/z 89.04, were detected from all the derivatives by the collision induced dissociation of the protonated molecular ions. A highly sensitive detection at a low-fmol level (0.5–3.2 fmol) was also obtained from the selected reaction monitoring (SRM) chromatograms. An isotope labeling strategy using light and heavy L-PGA-OSu for the differential analysis of the DL-amino acids in different sample groups is also presented in this paper. The differential analysis of biological sample (i.e., human serum) and food product (i.e., yogurt) were tried to demonstrate the efficiency of the proposed method. The ratios of the DL-amino acids in human serum samples, spiked with the different concentrations of D-amino acids, were determined by the procedures using L-PGA-OSu and L-PGA[d 5 ]-OSu. The D/L ratios in the two sample groups at different concentrations of amino

Energetics of the molecular interactions of L-alanine and L-serine with xylitol, D-sorbitol, and D-mannitol in aqueous solutions at 298.15 K

Science.gov (United States)

Mezhevoi, I. N.; Badelin, V. G.

2013-04-01

Integral enthalpies of dissolution Δsol H m of L-alanine and L-serine are measured via the calorimetry of dissolution in aqueous solutions of xylitol, D-sorbitol, and D-mannitol. Standard enthalpies of dissolution (Δsol H ○) and the transfer (Δtr H ○) of amino acids from water to binary solvent are calculated from the experimental data. Using the McMillan-Mayer theory, enthalpy coefficients of pairwise interactions h xy of amino acids with molecules of polyols are calculated that are negative. The obtained results are discussed within the theory of the prevalence of different types of interactions in mixed solutions and the effect of the structural features of interacting biomolecules on the thermochemical parameters of dissolution of amino acids.

Dissociation constants and thermodynamic properties of amino acids used in CO2 absorption from (293 to 353) K

NARCIS (Netherlands)

Hamborg, E. S.; Niederer, J. P. M.; Versteeg, G. F.

2007-01-01

The second dissociation constants of the amino acids βalanine, taurine, sarcosine, 6-aminohexanoic acid, DL-methionine, glycine, L-phenylalanine, and L-proline and the third dissociation constants of L-glutamic acid and L-aspartic acid have been determined from electromotive force measurements at

Dissociation Constants and Thermodynamic Properties of Amino Acids Used in CO2 Absorption from (293 to 353) K

NARCIS (Netherlands)

Hamborg, Espen; Niederer, John; Versteeg, Geert

2007-01-01

The second dissociation constants of the amino acids β-alanine, taurine, sarcosine, 6-aminohexanoic acid, dl-methionine, glycine, l-phenylalanine, and l-proline and the third dissociation constants of l-glutamic acid and l-aspartic acid have been determined from electromotive force measurements at

Responses of garlic bulbs to gamma irradiation. Changes in major amino acids

International Nuclear Information System (INIS)

Parolo, Maria E.; Orioli, Gustavo A.; Croci, Clara A.

1997-01-01

Studies were conducted to provide information about the amino acids composition of garlic bulbs cv Colorado and to determinate the effect of a dose of 60 Gy of gamma rays on the behavior of the major free amino acids in relation to sprout growth radioinhibition. TLC and HPLC were used for identification and quantification of free amino acids. Eighteen free amino acids were identified in both parts of garlic bulbs: alanine, glycine, proline, methionine, serine, phenylalanine, aspartic acid, glutamic acid, lysine, glutamine, arginine, tyrosine, threonine, cystine, cysteine, leucine + isoleucine and asparagine. In the inner sprout the major amino acids founded were: glutamine, glutamic acid, threonine, asparagine, cystine, cysteine and methionine; in the storage leaf also arginine was also predominant. In general concentration of amino acids appeared to less affected by irradiation in the storage leaf that in the inner sprout. An increase in the short time post-irradiation in glutamine, glutamic acid, asparagine, theorine and methionine was observed. Sprout grouth radioinhibition was evident about 70 days after treatment and was preceded by a decrease in the major amino acids except methionine. It appears that concentration of same major amino acidscan be used as monitors of radioinhibition process in inner sprout of garlic. (author). 15 refs., 8 figs

Contributions of basic amino acids in the autolysis loop of factor XIa to serpin specificity.

Science.gov (United States)

Rezaie, Alireza R; Sun, Mao-fu; Gailani, David

2006-08-08

The autolysis loops (amino acids 143-154, chymotrypsinogen numbering) of plasma serine proteases play key roles in determining the specificity of protease inhibition by plasma serpins. We studied the importance of four basic residues (Arg-144, Lys-145, Arg-147, and Lys-149) in the autolysis loop of the coagulation protease factor XIa (fXIa) for inhibition by serpins. Recombinant fXIa mutants, in which these residues were replaced individually or in combination with alanine, were prepared. The proteases were compared to wild-type fXIa (fXIa-WT) with respect to their ability to activate factor IX in a plasma clotting assay, to hydrolyze the chromogenic substrate S2366, and to undergo inhibition by the C1-inhibitor (C1-INH), protein Z dependent protease inhibitor (ZPI), antithrombin (AT), and alpha(1)-protease inhibitor (alpha(1)-PI). All mutants exhibited normal activity in plasma and hydrolyzed S2366 with catalytic efficiencies similar to that of fXIa-WT. Inhibition of mutants by C1-INH was increased to varying degrees relative to that of fXIa-WT, with the mutant containing alanine replacements for all four basic residues (fXIa-144-149A) exhibiting an approximately 15-fold higher rate of inhibition. In contrast, the inhibition by ZPI was impaired 2-3-fold for single amino acid substitutions, and fXIa-144-149A was essentially resistant to inhibition by ZPI. Alanine substitution for Arg-147 impaired inhibition by AT approximately 7-fold; however, other substitutions did not affect it or slightly enhanced inhibition. Arg-147 was also required for inhibition by alpha(1)-PI. Cumulatively, the results demonstrate that basic amino acids in the autolysis loop of fXIa are important determinants of serpin specificity.

Characterization of the l-alanine exporter AlaE of Escherichia coli and its potential role in protecting cells from a toxic-level accumulation of l-alanine and its derivatives.

Science.gov (United States)

Kim, Seryoung; Ihara, Kohei; Katsube, Satoshi; Hori, Hatsuhiro; Ando, Tasuke; Isogai, Emiko; Yoneyama, Hiroshi

2015-08-01

We previously reported that the alaE gene of Escherichia coli encodes the l-alanine exporter AlaE. The objective of this study was to elucidate the mechanism of the AlaE exporter. The minimum inhibitory concentration of l-alanine and l-alanyl-l-alanine in alaE-deficient l-alanine-nonmetabolizing cells MLA301ΔalaE was 4- and >4000-fold lower, respectively, than in the alaE-positive parent cells MLA301, suggesting that AlaE functions as an efflux pump to avoid a toxic-level accumulation of intracellular l-alanine and its derivatives. Furthermore, the growth of the alaE-deficient mutant derived from the l-alanine-metabolizing strain was strongly inhibited in the presence of a physiological level of l-alanyl-l-alanine. Intact MLA301ΔalaE and MLA301ΔalaE/pAlaE cells producing plasmid-borne AlaE, accumulated approximately 200% and 50%, respectively, of the [(3) H]l-alanine detected in MLA301 cells, suggesting that AlaE exports l-alanine. When 200 mmol/L l-alanine-loaded inverted membrane vesicles prepared from MLA301ΔalaE/pAlaE were placed in a solution containing 200 mmol/L or 0.34 μmol/L l-alanine, energy-dependent [(3) H]l-alanine accumulation occurred under either condition. This energy-dependent uphill accumulation of [(3) H]l-alanine was strongly inhibited in the presence of carbonyl cyanide m-chlorophenylhydrazone but not by dicyclohexylcarbodiimide, suggesting that the AlaE-mediated l-alanine extrusion was driven by proton motive force. Based on these results, physiological roles of the l-alanine exporter are discussed. © 2015 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.

EPR/alanine dosimetry for two therapeutic proton beams

Energy Technology Data Exchange (ETDEWEB)

Marrale, Maurizio, E-mail: maurizio.marrale@unipa.it [Dipartimento di Fisica e Chimica, Università di Palermo, Viale delle Scienze, Edificio 18, 90128 Palermo (Italy); Gruppo V Sezione INFN di Catania, Via Santa Sofia, 64, 95123 Catania (Italy); Carlino, Antonio [Dipartimento di Fisica e Chimica, Università di Palermo, Viale delle Scienze, Edificio 18, 90128 Palermo (Italy); EBG MedAustron GmbH, Marie Curie-Straße 5, A-2700 Wiener Neustadt (Austria); Gallo, Salvatore [Dipartimento di Fisica e Chimica, Università di Palermo, Viale delle Scienze, Edificio 18, 90128 Palermo (Italy); Gruppo V Sezione INFN di Catania, Via Santa Sofia, 64, 95123 Catania (Italy); Laboratorio PH3DRA, Dipartimento di Fisica e Astronomia, Università di Catania, Via Santa Sofia 64, 95123 Catania (Italy); Longo, Anna; Panzeca, Salvatore [Dipartimento di Fisica e Chimica, Università di Palermo, Viale delle Scienze, Edificio 18, 90128 Palermo (Italy); Gruppo V Sezione INFN di Catania, Via Santa Sofia, 64, 95123 Catania (Italy); Bolsi, Alessandra; Hrbacek, Jan; Lomax, Tony [Center for Proton Therapy, Paul Scherrer Institute, 5232 Villigen PSI (Switzerland)

2016-02-01

In this work the analysis of the electron paramagnetic resonance (EPR) response of alanine pellets exposed to two different clinical proton beams employed for radiotherapy is performed. One beam is characterized by a passive delivery technique and is dedicated to the eyes treatment (OPTIS2 beam line). Alanine pellets were irradiated with a 70 MeV proton beam corresponding to 35 mm range in eye tissue. We investigated how collimators with different sizes and shape used to conform the dose to the planned target volume influence the delivered dose. For this purpose we performed measurements with varying the collimator size (Output Factor) and the results were compared with those obtained with other dosimetric techniques (such as Markus chamber and diode detector). This analysis showed that the dosimeter response is independent of collimator diameter if this is larger than or equal to 10 mm. The other beam is characterized by an active spot-scanning technique, the Gantry1 beam line (maximum energy 230 MeV), and is used to treat deep-seated tumors. The dose linearity of alanine response in the clinical dose range was tested and the alanine dose response at selected locations in depth was measured and compared with the TPS planned dose in a quasi-clinical scenario. The alanine response was found to be linear in the dose in the clinical explored range (from 10 to 70 Gy). Furthermore, a depth dose profile in a quasi-clinical scenario was measured and compared to the dose computed by the Treatment Planning System PSIPLAN. The comparison of calibrated proton alanine measurements and TPS dose shows a difference under 1% in the SOBP and a “quenching” effect up to 4% in the distal part of SOBP. The positive dosimetric characteristics of the alanine pellets confirm the feasibility to use these detectors for “in vivo” dosimetry in clinical proton beams.

EPR/alanine dosimetry for two therapeutic proton beams

International Nuclear Information System (INIS)

Marrale, Maurizio; Carlino, Antonio; Gallo, Salvatore; Longo, Anna; Panzeca, Salvatore; Bolsi, Alessandra; Hrbacek, Jan; Lomax, Tony

2016-01-01

In this work the analysis of the electron paramagnetic resonance (EPR) response of alanine pellets exposed to two different clinical proton beams employed for radiotherapy is performed. One beam is characterized by a passive delivery technique and is dedicated to the eyes treatment (OPTIS2 beam line). Alanine pellets were irradiated with a 70 MeV proton beam corresponding to 35 mm range in eye tissue. We investigated how collimators with different sizes and shape used to conform the dose to the planned target volume influence the delivered dose. For this purpose we performed measurements with varying the collimator size (Output Factor) and the results were compared with those obtained with other dosimetric techniques (such as Markus chamber and diode detector). This analysis showed that the dosimeter response is independent of collimator diameter if this is larger than or equal to 10 mm. The other beam is characterized by an active spot-scanning technique, the Gantry1 beam line (maximum energy 230 MeV), and is used to treat deep-seated tumors. The dose linearity of alanine response in the clinical dose range was tested and the alanine dose response at selected locations in depth was measured and compared with the TPS planned dose in a quasi-clinical scenario. The alanine response was found to be linear in the dose in the clinical explored range (from 10 to 70 Gy). Furthermore, a depth dose profile in a quasi-clinical scenario was measured and compared to the dose computed by the Treatment Planning System PSIPLAN. The comparison of calibrated proton alanine measurements and TPS dose shows a difference under 1% in the SOBP and a “quenching” effect up to 4% in the distal part of SOBP. The positive dosimetric characteristics of the alanine pellets confirm the feasibility to use these detectors for “in vivo” dosimetry in clinical proton beams.

Alanine repeats influence protein localization in splicing speckles and paraspeckles.

Science.gov (United States)

Chang, Shuo-Hsiu; Chang, Wei-Lun; Lu, Chia-Chen; Tarn, Woan-Yuh

2014-12-16

Mammalian splicing regulatory protein RNA-binding motif protein 4 (RBM4) has an alanine repeat-containing C-terminal domain (CAD) that confers both nuclear- and splicing speckle-targeting activities. Alanine-repeat expansion has pathological potential. Here we show that the alanine-repeat tracts influence the subnuclear targeting properties of the RBM4 CAD in cultured human cells. Notably, truncation of the alanine tracts redistributed a portion of RBM4 to paraspeckles. The alanine-deficient CAD was sufficient for paraspeckle targeting. On the other hand, alanine-repeat expansion reduced the mobility of RBM4 and impaired its splicing activity. We further took advantage of the putative coactivator activator (CoAA)-RBM4 conjoined splicing factor, CoAZ, to investigate the function of the CAD in subnuclear targeting. Transiently expressed CoAZ formed discrete nuclear foci that emerged and subsequently separated-fully or partially-from paraspeckles. Alanine-repeat expansion appeared to prevent CoAZ separation from paraspeckles, resulting in their complete colocalization. CoAZ foci were dynamic but, unlike paraspeckles, were resistant to RNase treatment. Our results indicate that the alanine-rich CAD, in conjunction with its conjoined RNA-binding domain(s), differentially influences the subnuclear localization and biogenesis of RBM4 and CoAZ. © The Author(s) 2014. Published by Oxford University Press on behalf of Nucleic Acids Research.

The result of Alanine/ESR dosimetry at Wolsung unit 1

International Nuclear Information System (INIS)

Park, Byeong Ryong; Choi, Hoon; Lim, Young Khi

2008-01-01

It needs accurate estimation of radiation level for verifying machinery and cable in Nuclear Power Plant. Therefore, in this study, we used ESR(Electron Spin Resonance) system for estimate dose of Alanine dosimeter. Alanine/ESR dosimetry, already known as a dosimetric method in medical and industrial field, was applied to estimate dose quantity at cable locations within a nuclear power plant as a part of equipment qualification program. Alanine/ESR dosimetry of absorbed dose range is 1 - 100 KGy. The alanine dosimeter is not significantly affected by temperature and fading is limited to 1% per year. The alanine dosimeters were fixed on the targeted cable or nearest position to measure dose quantity to get accurate value. Alanine dosimeters were scanned by commercially used two different ESR systems, e-scan and EMX series for alanine dosimeters. To estimate more accurate dose, two environmental correction factors, irradiation temperature and dosimeter weight, were used in calculation of absorbed dose quantity. In this study, dose values which are alinine dosimeter from Wolsong unit 1 are measured by two ESR systems. And then the results was compared each other

L-alanine as a precursor of ethylamine in camellia sinensis

International Nuclear Information System (INIS)

Takeo, Tadakazu

1975-01-01

After absorption of ammonium nitrogen, nitrogen-deficient Camellia sinensis synthesized theanine following synthesis of glutamic acid and alanine. The rate of incorporation of 14 C from L-alanine U- 14 C into theanine was faster than from acetaldehyde 1-2 14 C. Incorporation of 14 C from L-alanine U- 14 C into the ethylamide of theanine was prevented by adding an excess of ethylamine to the culture solution. Green seedlings converted alanine to ethylamine more rapidly than did etiolated seedlings. (auth.)

Potential transfer of neurotoxic amino acid β-N-methylamino-alanine (BMAA) from mother to infant during breast-feeding: Predictions from human cell lines

International Nuclear Information System (INIS)

Andersson, Marie; Ersson, Lisa; Brandt, Ingvar; Bergström, Ulrika

2017-01-01

β-N-methylamino-alanine (BMAA) is a non-protein amino acid produced by cyanobacteria, diatoms and dinoflagellates. BMAA has potential to biomagnify in a terrestrial food chain, and to bioaccumulate in fish and shellfish. We have reported that administration of [ 14 C]L-BMAA to lactating mice and rats results in a mother to off-spring transfer via the milk. A preferential enantiomer-specific uptake of [ 14 C]L-BMAA has also been demonstrated in differentiated murine mammary epithelium HC11 cells. These findings, together with neurotoxic effects of BMAA demonstrated both in vitro and in vivo, highlight the need to determine whether such transfer could also occur in humans. Here, we used four cell lines of human origin to examine and compare the transport of the two BMAA enantiomers in vitro. The uptake patterns of [ 14 C]L- and [ 14 C]D-BMAA in the human mammary MCF7 cell line were in agreement with the results in murine HC11 cells, suggesting a potential secretion of BMAA into human breast milk. The permeability coefficients for both [ 14 C]L- and [ 14 C]D-BMAA over monolayers of human intestinal Caco2 cells supported an efficient absorption from the human intestine. As a final step, transport experiments confirmed that [ 14 C]L-and [ 14 C]D-BMAA can be taken up by human SHSY5Y neuroblastoma cells and even more efficiently by human U343 glioblastoma cells. In competition experiments with various amino acids, the ASCT2 specific inhibitor benzylserine was the most effective inhibitor of [ 14 C]L-BMAA uptake tested here. Altogether, our results suggest that BMAA can be transferred from an exposed mother, via the milk, to the brain of the nursed infant. - Highlights: • Transport of BMAA in human intestinal, mammary and CNS cell lines was examined. • The transport of L-BMAA over intestinal cell monolayers was unidirectional. • Enantiomer-selective uptake of L-BMAA in breast, neuron and glia cells was evident. • Competition experiments indicate that L-BMAA uptake

Potential transfer of neurotoxic amino acid β-N-methylamino-alanine (BMAA) from mother to infant during breast-feeding: Predictions from human cell lines

Energy Technology Data Exchange (ETDEWEB)

Andersson, Marie [Department of Environmental Toxicology, Uppsala University, Norbyvägen 18A, SE-752 36 Uppsala (Sweden); Ersson, Lisa [Department of Pharmaceutical Biosciences, Uppsala University, Box 591, SE-751 24 Uppsala (Sweden); Brandt, Ingvar, E-mail: Ingvar.Brandt@ebc.uu.se [Department of Environmental Toxicology, Uppsala University, Norbyvägen 18A, SE-752 36 Uppsala (Sweden); Bergström, Ulrika [Department of Environmental Toxicology, Uppsala University, Norbyvägen 18A, SE-752 36 Uppsala (Sweden); Swedish Defence Research Agency, Division of CBRN Defence and Security, SE-164 90 Stockholm (Sweden)

2017-04-01

β-N-methylamino-alanine (BMAA) is a non-protein amino acid produced by cyanobacteria, diatoms and dinoflagellates. BMAA has potential to biomagnify in a terrestrial food chain, and to bioaccumulate in fish and shellfish. We have reported that administration of [{sup 14}C]L-BMAA to lactating mice and rats results in a mother to off-spring transfer via the milk. A preferential enantiomer-specific uptake of [{sup 14}C]L-BMAA has also been demonstrated in differentiated murine mammary epithelium HC11 cells. These findings, together with neurotoxic effects of BMAA demonstrated both in vitro and in vivo, highlight the need to determine whether such transfer could also occur in humans. Here, we used four cell lines of human origin to examine and compare the transport of the two BMAA enantiomers in vitro. The uptake patterns of [{sup 14}C]L- and [{sup 14}C]D-BMAA in the human mammary MCF7 cell line were in agreement with the results in murine HC11 cells, suggesting a potential secretion of BMAA into human breast milk. The permeability coefficients for both [{sup 14}C]L- and [{sup 14}C]D-BMAA over monolayers of human intestinal Caco2 cells supported an efficient absorption from the human intestine. As a final step, transport experiments confirmed that [{sup 14}C]L-and [{sup 14}C]D-BMAA can be taken up by human SHSY5Y neuroblastoma cells and even more efficiently by human U343 glioblastoma cells. In competition experiments with various amino acids, the ASCT2 specific inhibitor benzylserine was the most effective inhibitor of [{sup 14}C]L-BMAA uptake tested here. Altogether, our results suggest that BMAA can be transferred from an exposed mother, via the milk, to the brain of the nursed infant. - Highlights: • Transport of BMAA in human intestinal, mammary and CNS cell lines was examined. • The transport of L-BMAA over intestinal cell monolayers was unidirectional. • Enantiomer-selective uptake of L-BMAA in breast, neuron and glia cells was evident. • Competition

Amino Acid Signatures to Evaluate the Beneficial Effects of Weight Loss

Directory of Open Access Journals (Sweden)

Nina Geidenstam

2017-01-01

Full Text Available Aims. We investigated the relationship between circulating amino acid levels and obesity; to what extent weight loss followed by weight maintenance can correct amino acid abnormalities; and whether amino acids are related to weight loss. Methods. Amino acids associated with waist circumference (WC and BMI were studied in 804 participants from the Malmö Diet and Cancer Cardiovascular Cohort (MDC-CC. Changes in amino acid levels were analyzed after weight loss and weight maintenance in 12 obese subjects and evaluated in a replication cohort (n=83. Results. Out of the eight identified BMI-associated amino acids from the MDC-CC, alanine, isoleucine, tyrosine, phenylalanine, and glutamate decreased after weight loss, while asparagine increased after weight maintenance. These changes were validated in the replication cohort. Scores that were constructed based on obesity-associated amino acids and known risk factors decreased in the ≥10% weight loss group with an associated change in BMI (R2=0.16–0.22, p<0.002, whereas the scores increased in the <10% weight loss group (p<0.0004. Conclusions. Weight loss followed by weight maintenance leads to differential changes in amino acid levels associated with obesity. Treatment modifiable scores based on epidemiological and interventional data may be used to evaluate the potential metabolic benefit of weight loss.

Analysis of an Alanine/Arginine Mixture by Using TLC/FTIR Technique

Directory of Open Access Journals (Sweden)

Jun Liu

2014-01-01

Full Text Available We applied TLC/FTIR coupled with mapping technique to analyze an alanine/arginine mixture. Narrow band TLC plates prepared by using AgI as a stationary phase were used to separate alanine and arginine. The distribution of alanine and arginine spots was manifested by a 3D chromatogram. Alanine and arginine can be successfully separated by the narrow band TLC plate. In addition, the FTIR spectra of the separated alanine and arginine spots on the narrow band TLC plate are roughly the same as the corresponding reference IR spectra.

Environmentally safe corrosion inhibition of Mg-Al-Zn alloy in chloride free neutral solutions by amino acids

Energy Technology Data Exchange (ETDEWEB)

Helal, N.H. [Chemistry Department, Faculty of Science, Fayoum University, Fayoum (Egypt); Badawy, W.A., E-mail: wbadawy@cu.edu.eg [Chemistry Department, Faculty of Science, Cairo University, 12 613 Giza (Egypt)

2011-07-30

Highlights: > Phenyl alanine at a concentration of 2 x 10{sup -3} mol dm{sup -3} gives 93% corrosion inhibition efficiency for the corrosion of the Mg-Al-Zn alloy. > The corrosion inhibition process is based on the adsorption of the amino acid molecules on the active sites of the alloy surface by physical adsorption mechanism. > The adsorption free energy was 15.72 kJ mol{sup -1}. - Abstract: The corrosion inhibition of Mg-Al-Zn alloy was investigated in stagnant naturally aerated chloride free neutral solutions using amino acids as environmentally safe corrosion inhibitors. The corrosion rate was calculated in the absence and presence of the corrosion inhibitor using the polarization technique and electrochemical impedance spectroscopy. The experimental impedance data were fitted to theoretical data according to a proposed electronic circuit model to explain the behavior of the alloy/electrolyte interface under different conditions. The corrosion inhibition process was found to depend on the adsorption of the amino acid molecules on the metal surface. Phenyl alanine has shown remarkably high corrosion inhibition efficiency up to 93% at a concentration of 2 x 10{sup -3} mol dm{sup -3}. The corrosion inhibition efficiency was found to depend on the concentration of the amino acid and its structure. The mechanism of the corrosion inhibition process was discussed and different adsorption isotherms were investigated. The free energy of the adsorption process was calculated for the adsorption of different amino acids on the Mg-Al-Zn alloy and the obtained values reveal a physical adsorption of the inhibitor molecules on the alloy surface.

Volumetric properties of l-alanine, and l-valine in aqueous sucrose solutions at T=(288.15 and 308.15) K

International Nuclear Information System (INIS)

Pal, Amalendu; Kumar, Suresh

2005-01-01

Densities of l-alanine, and l-valine have been measured at T=(288.15 and 308.15) K in aqueous sucrose solutions ranging from pure water to 25 mass% of sucrose. From these densities, apparent molar volumes (V φ ) and limiting partial molar volumes (V φ 0 ) of each amino acid in various aqueous sucrose solutions have been evaluated. These data were combined with the earlier reported V φ 0 values of l-alanine, and l-valine in aqueous sucrose solutions at T=298.15 K to calculate the (-bar V φ 0 /-bar T) P values. The partial molar properties of transfer (ΔV φ 0 ) from water to aqueous sucrose solutions at infinite dilution has been calculated. Transfer parameters have been interpreted in terms of solute-cosolute interactions on the basis of a cosphere overlap model. Pair and triplet interaction coefficients have also been calculated from transfer volume data

Flux Analysis of Free Amino Sugars and Amino Acids in Soils by Isotope Tracing with a Novel Liquid Chromatography/High Resolution Mass Spectrometry Platform.

Science.gov (United States)

Hu, Yuntao; Zheng, Qing; Wanek, Wolfgang

2017-09-05

Soil fluxomics analysis can provide pivotal information for understanding soil biochemical pathways and their regulation, but direct measurement methods are rare. Here, we describe an approach to measure soil extracellular metabolite (amino sugar and amino acid) concentrations and fluxes based on a 15 N isotope pool dilution technique via liquid chromatography and high-resolution mass spectrometry. We produced commercially unavailable 15 N and 13 C labeled amino sugars and amino acids by hydrolyzing peptidoglycan isolated from isotopically labeled bacterial biomass and used them as tracers ( 15 N) and internal standards ( 13 C). High-resolution (Orbitrap Exactive) MS with a resolution of 50 000 allowed us to separate different stable isotope labeled analogues across a large range of metabolites. The utilization of 13 C internal standards greatly improved the accuracy and reliability of absolute quantification. We successfully applied this method to two types of soils and quantified the extracellular gross fluxes of 2 amino sugars, 18 amino acids, and 4 amino acid enantiomers. Compared to the influx and efflux rates of most amino acids, similar ones were found for glucosamine, indicating that this amino sugar is released through peptidoglycan and chitin decomposition and serves as an important nitrogen source for soil microorganisms. d-Alanine and d-glutamic acid derived from peptidoglycan decomposition exhibited similar turnover rates as their l-enantiomers. This novel approach offers new strategies to advance our understanding of the production and transformation pathways of soil organic N metabolites, including the unknown contributions of peptidoglycan and chitin decomposition to soil organic N cycling.

Flux Analysis of Free Amino Sugars and Amino Acids in Soils by Isotope Tracing with a Novel Liquid Chromatography/High Resolution Mass Spectrometry Platform

Science.gov (United States)

2017-01-01

Soil fluxomics analysis can provide pivotal information for understanding soil biochemical pathways and their regulation, but direct measurement methods are rare. Here, we describe an approach to measure soil extracellular metabolite (amino sugar and amino acid) concentrations and fluxes based on a 15N isotope pool dilution technique via liquid chromatography and high-resolution mass spectrometry. We produced commercially unavailable 15N and 13C labeled amino sugars and amino acids by hydrolyzing peptidoglycan isolated from isotopically labeled bacterial biomass and used them as tracers (15N) and internal standards (13C). High-resolution (Orbitrap Exactive) MS with a resolution of 50 000 allowed us to separate different stable isotope labeled analogues across a large range of metabolites. The utilization of 13C internal standards greatly improved the accuracy and reliability of absolute quantification. We successfully applied this method to two types of soils and quantified the extracellular gross fluxes of 2 amino sugars, 18 amino acids, and 4 amino acid enantiomers. Compared to the influx and efflux rates of most amino acids, similar ones were found for glucosamine, indicating that this amino sugar is released through peptidoglycan and chitin decomposition and serves as an important nitrogen source for soil microorganisms. d-Alanine and d-glutamic acid derived from peptidoglycan decomposition exhibited similar turnover rates as their l-enantiomers. This novel approach offers new strategies to advance our understanding of the production and transformation pathways of soil organic N metabolites, including the unknown contributions of peptidoglycan and chitin decomposition to soil organic N cycling. PMID:28776982

Determination of Theophylline Binding to Human Serum Proteins by Isotachophoresis

NARCIS (Netherlands)

Reijenga, J.C.; Gaijkema, A.P.M.; Mikkers, F.E.P.

1984-01-01

Free theophylline was isolated from human serum by ultrafiltration and analysed in a leading electrolyte of 7.5 mM morpholinoethanesulphoric acid with ammediol as a counter ion at pH 8.90 and -alanine as a terminator. The UV (280 nm) absorbance of the theophylline spike between serine and bicine as

EPR of alanine irradiated by neutrons

International Nuclear Information System (INIS)

Pivovarov, S.P.; Seredavina, T.A.; Zhdanov, S.V.; Mul'gin, S.I.; Zhakparov, R.K.

2001-01-01

In the work the first results of EPR studies of alanine, irradiated with diverse doses at neutron cyclotron generator different conditions and on the critical reactor stand are presented. A dose linearity dependence of EPR signal is observing, the methods of γ-background contribution separation are discussed. Obtain results is giving the basis to recommendation of alanine as an effective detector irradiation. However it is demanded the farther study on clarification of radiation sensitivity value dependence on the neutron energy spectrum form

1H NMR-based serum metabolomics reveals erythromycin-induced liver toxicity in albino Wistar rats

Directory of Open Access Journals (Sweden)

Atul Rawat

2016-01-01

Full Text Available Introduction: Erythromycin (ERY is known to induce hepatic toxicity which mimics other liver diseases. Thus, ERY is often used to produce experimental models of drug-induced liver-toxicity. The serum metabolic profiles can be used to evaluate the liver-toxicity and to further improve the understanding of underlying mechanism. Objective: To establish the serum metabolic patterns of Erythromycin induced hepatotoxicity in albino wistar rats using 1H NMR based serum metabolomics. Experimental: Fourteen male rats were randomly divided into two groups (n = 7 in each group: control and ERY treated. After 28 days of intervention, the metabolic profiles of sera obtained from ERY and control groups were analyzed using high-resolution 1D 1H CPMG and diffusion-edited nuclear magnetic resonance (NMR spectra. The histopathological and SEM examinations were employed to evaluate the liver toxicity in ERY treated group. Results: The serum metabolic profiles of control and ERY treated rats were compared using multivariate statistical analysis and the metabolic patterns specific to ERY-induced liver toxicity were established. The toxic response of ERY was characterized with: (a increased serum levels of Glucose, glutamine, dimethylamine, malonate, choline, phosphocholine and phospholipids and (b decreased levels of isoleucine, leucine, valine, alanine, glutamate, citrate, glycerol, lactate, threonine, circulating lipoproteins, N-acetyl glycoproteins, and poly-unsaturated lipids. These metabolic alterations were found to be associated with (a decreased TCA cycle activity and enhanced fatty acid oxidation, (b dysfunction of lipid and amino acid metabolism and (c oxidative stress. Conclusion and Recommendations: Erythromycin is often used to produce experimental models of liver toxicity; therefore, the established NMR-based metabolic patterns will form the basis for future studies aiming to evaluate the efficacy of anti-hepatotoxic agents or the hepatotoxicity of new

Plasma amino acid levels are elevated in young, healthy low birth weight men exposed to short-term high-fat overfeeding

DEFF Research Database (Denmark)

Ribel-Madsen, Amalie; Hellgren, Lars; Brøns, Charlotte

2016-01-01

Low birth weight (LBW) individuals exhibit a disproportionately increased, incomplete fatty acid oxidation and a decreased glucose oxidation, compared with normal birth weight (NBW) individuals, and furthermore have an increased risk of developing insulin resistance and type 2 diabetes. We...... and after a 5‐day high‐fat, high‐calorie diet. We demonstrated that LBW and NBW men increased plasma alanine levels and decreased valine and leucine/isoleucine levels in response to overfeeding. Also, LBW men had higher alanine, proline, methionine, citrulline, and total amino acid levels after overfeeding...

Graphene for amino acid biosensing: Theoretical study of the electronic transport

Science.gov (United States)

Rodríguez, S. J.; Makinistian, L.; Albanesi, E. A.

2017-10-01

The study of biosensors based on graphene has increased in the last years, the combination of excellent electrical properties and low noise makes graphene a material for next generation electronic devices. This work discusses the application of a graphene-based biosensor for the detection of amino acids histidine (His), alanine (Ala), aspartic acid (Asp), and tyrosine (Tyr). First, we present the results of modeling from first principles the adsorption of the four amino acids on a graphene sheet, we calculate adsorption energy, substrate-adsorbate distance, equilibrium geometrical configurations (upon relaxation) and densities of states (DOS) for each biomolecule adsorbed. Furthermore, in order to evaluate the effects of amino acid adsorption on the electronic transport of graphene, we modeled a device using first-principles calculations with a combination of Density Functional Theory (DFT) and Nonequilibrium Greens Functions (NEGF). We provide with a detailed discussion in terms of transmission, current-voltage curves, and charge transfer. We found evidence of differences in the electronic transport through the graphene sheet due to amino acid adsorption, reinforcing the possibility of graphene-based sensors for amino acid sequencing of proteins.

Adsorption behavior of oxidized galactomannans onto amino terminated surfaces and their interaction with bovine serum albumin

International Nuclear Information System (INIS)

Sierakowski, M.-R; Silva, Maria R.V. da; Freitas, R.A.; Moreira, Jose S.R.; Fujimoto, J.; Petri, D.F.S.; Cordeiro, Paulo R.D.; Andrade, Fabiana D.

2001-01-01

A galactomannan (CF) extracted from Cassia fastuosa seeds was purified and oxidized with (2,2,6,6- tetramethylpiperidine-1-oxyl) to form a uronic acid-containing polysaccharide (CFOX) with a degree of oxidation (DO) of 0.22. The chemical structures of CF and CFOX were characterized. The adsorption behavior of CF and CFOX onto amino-terminated surfaces was studied by means of ellipsometric measurements. The influence of p H and ionic strength on the adsorption was also investigated. At p H 4, there was a maximum in the adsorbed amount caused by strong electrostatic attraction between the substrate and the oxidized galactomannans. There was no ionic strength effect on the adsorption behavior. The immobilization of bovine serum albumin onto CF and CFOX was studied as a function of p H. At the isoelectric point a maximum in the adsorbed amount was found. (author)

Adsorption behavior of oxidized galactomannans onto amino terminated surfaces and their interaction with bovine serum albumin

Energy Technology Data Exchange (ETDEWEB)

Sierakowski, M.-R; Silva, Maria R.V. da [Universidade Federal do Parana, Curitiba, PR (Brazil). Dept. de Quimica. Lab. de Biopolimeros]. E-mail: mrbiopol@quimica.ufpr.br; Freitas, R.A.; Moreira, Jose S.R. [Universidade Federal do Parana, Curitiba, PR (Brazil). Dept. de Bioquimica; Fujimoto, J.; Petri, D.F.S.; Cordeiro, Paulo R.D. [Sao Paulo Univ., SP (Brazil). Inst. de Quimica]. E-mail: dfsp@quim.iq.usp.br; Andrade, Fabiana D

2001-07-01

A galactomannan (CF) extracted from Cassia fastuosa seeds was purified and oxidized with (2,2,6,6- tetramethylpiperidine-1-oxyl) to form a uronic acid-containing polysaccharide (CFOX) with a degree of oxidation (DO) of 0.22. The chemical structures of CF and CFOX were characterized. The adsorption behavior of CF and CFOX onto amino-terminated surfaces was studied by means of ellipsometric measurements. The influence of p H and ionic strength on the adsorption was also investigated. At p H 4, there was a maximum in the adsorbed amount caused by strong electrostatic attraction between the substrate and the oxidized galactomannans. There was no ionic strength effect on the adsorption behavior. The immobilization of bovine serum albumin onto CF and CFOX was studied as a function of p H. At the isoelectric point a maximum in the adsorbed amount was found. (author)

Alanine EPR dosimeter response in proton therapy beams

International Nuclear Information System (INIS)

Gall, K.; Serago, C.; Desrosiers, M.; Bensen, D.

1997-01-01

We report a series of measurements directed to assess the suitability of alanine as a mailable dosimeter for dosimetry quality assurance of proton radiation therapy beams. These measurements include dose-response of alanine at 140 MeV, and comparison of response vs energy with a parallel plate ionization chamber. All irradiations were made at the Harvard Cyclotron Laboratory, and the dosimeters were read at NIST. The results encourage us that alanine could be expected to serve as a mailable dosimeter with systematic error due to differential energy response no greater than 3% when doses of 25 Gy are used. (Author)

Reversible uptake of molecular oxygen by heteroligand Co(II)-L-α-amino acid-imidazole systems: equilibrium models at full mass balance.

Science.gov (United States)

Pająk, Marek; Woźniczka, Magdalena; Vogt, Andrzej; Kufelnicki, Aleksander

2017-09-19

The paper examines Co(II)-amino acid-imidazole systems (where amino acid = L-α-amino acid: alanine, asparagine, histidine) which, when in aqueous solutions, activate and reversibly take up dioxygen, while maintaining the structural scheme of the heme group (imidazole as axial ligand and O 2 uptake at the sixth, trans position) thus imitating natural respiratory pigments such as myoglobin and hemoglobin. The oxygenated reaction shows higher reversibility than for Co(II)-amac systems with analogous amino acids without imidazole. Unlike previous investigations of the heteroligand Co(II)-amino acid-imidazole systems, the present study accurately calculates all equilibrium forms present in solution and determines the [Formula: see text]equilibrium constants without using any simplified approximations. The equilibrium concentrations of Co(II), amino acid, imidazole and the formed complex species were calculated using constant data obtained for analogous systems under oxygen-free conditions. Pehametric and volumetric (oxygenation) studies allowed the stoichiometry of O 2 uptake reaction and coordination mode of the central ion in the forming oxygen adduct to be determined. The values of dioxygen uptake equilibrium constants [Formula: see text] were evaluated by applying the full mass balance equations. Investigations of oxygenation of the Co(II)-amino acid-imidazole systems indicated that dioxygen uptake proceeds along with a rise in pH to 9-10. The percentage of reversibility noted after acidification of the solution to the initial pH ranged within ca 30-60% for alanine, 40-70% for asparagine and 50-90% for histidine, with a rising tendency along with the increasing share of amino acid in the Co(II): amino acid: imidazole ratio. Calculations of the share of the free Co(II) ion as well as of the particular complex species existing in solution beside the oxygen adduct (regarding dioxygen bound both reversibly and irreversibly) indicated quite significant values for the

Alanine/ESR dosimetry system for routine use in radiation processing

International Nuclear Information System (INIS)

Kojima, T.; Haruyama, Y.; Tachibana, H.; Tanaka, R.; Okamoto, J.; Hara, H.; Kashiwazaki, S.

1993-01-01

A new alanine-polystyrene(PS) dosimeter prepared with simplified molding procedure and an automatic desk-top dose-reader of alanine dosimeter were developed for the purpose of routine use. Combination of these two allows us to apply a reliable alanine/ESR dosimetry system to routine dosimetric process control in industrial gamma radiation processing. (Author)

Radiation-induced increase in the release of amino acids by isolated, perfused skeletal muscle

International Nuclear Information System (INIS)

Schwenen, M.

1989-01-01

Local exposure of the hindquarter of the rat to 15Gy of gamma-radiation resulted, 4-6h after irradiation, in increased release of amino acids by the isolated, perfused hindquarter preparation, 70% of which is skeletal muscle. This increase in release involves not only alanine and glutamine, but also those amino acids not metabolized by muscle and, therefore, released in proportion to their occurrence in muscle proteins. Because metabolic parameters and content of energy-rich phosphate compounds in muscle remain unchanged, it is unlikely that general cellular damage is the underlying cause of the radiation-induced increase in amino acid release. The findings strongly favour the hypothesis that increased availability of amino acids results from enhanced protein break-down in skeletal muscle which has its onset shortly after irradiation. This radiation-induced disturbance in protein metabolism might be one of the pathogenetic factors in the aetiology of radiation myopathy. (author)

Isolation and characterization of racemase from Ensifer sp. 23-3 that acts on α-aminolactams and α-amino acid amides.

Science.gov (United States)

Matsui, Daisuke; Fuhshuku, Ken-Ichi; Nagamori, Shingo; Takata, Momoko; Asano, Yasuhisa

2017-11-01

Limited information is available on α-amino-ε-caprolactam (ACL) racemase (ACLR), a pyridoxal 5'-phosphate-dependent enzyme that acts on ACL and α-amino acid amides. In the present study, eight bacterial strains with the ability to racemize α-amino-ε-caprolactam were isolated and one of them was identified as Ensifer sp. strain 23-3. The gene for ACLR from Ensifer sp. 23-3 was cloned and expressed in Escherichia coli. The recombinant ACLR was then purified to homogeneity from the E. coli transformant harboring the ACLR gene from Ensifer sp. 23-3, and its properties were characterized. This enzyme acted not only on ACL but also on α-amino-δ-valerolactam, α-amino-ω-octalactam, α-aminobutyric acid amide, and alanine amide.

General synthesis of C-glycosyl amino acids via proline-catalyzed direct electrophilic alpha-amination of C-glycosylalkyl aldehydes.

Science.gov (United States)

Nuzzi, Andrea; Massi, Alessandro; Dondoni, Alessandro

2008-10-16

Non-natural axially and equatorially linked C-glycosyl alpha-amino acids (glycines, alanines, and CH2-serine isosteres) with either S or R alpha-configuration were prepared by D- and L-proline-catalyzed (de >95%) alpha-amination of C-glycosylalkyl aldehydes using dibenzyl azodicarboxylate as the electrophilic reagent.

PPARα regulates the hepatotoxic biomarker alanine aminotransferase (ALT1) gene expression in human hepatocytes

International Nuclear Information System (INIS)

Thulin, Petra; Rafter, Ingalill; Stockling, Kenneth; Tomkiewicz, Celine; Norjavaara, Ensio; Aggerbeck, Martine; Hellmold, Heike; Ehrenborg, Ewa; Andersson, Ulf; Cotgreave, Ian; Glinghammar, Bjoern

2008-01-01

In this work, we investigated a potential mechanism behind the observation of increased aminotransferase levels in a phase I clinical trial using a lipid-lowering drug, the peroxisome proliferator-activated receptor (PPAR) α agonist, AZD4619. In healthy volunteers treated with AZD4619, serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities were elevated without an increase in other markers for liver injury. These increases in serum aminotransferases have previously been reported in some patients receiving another PPARα agonist, fenofibrate. In subsequent in vitro studies, we observed increased expression of ALT1 protein and mRNA in human hepatocytes after treatment with fenofibric acid. The PPAR effect on ALT1 expression was shown to act through a direct transcriptional mechanism involving at least one PPAR response element (PPRE) in the proximal ALT1 promoter, while no effect of fenofibrate and AZD4619 was observed on the ALT2 promoter. Binding of PPARs to the PPRE located at - 574 bp from the transcriptional start site was confirmed on both synthetic oligonucleotides and DNA in hepatocytes. These data show that intracellular ALT expression is regulated by PPAR agonists and that this mechanism might contribute to increased ALT activity in serum

Effect of substituting arginine and lysine with alanine on antimicrobial activity and the mechanism of action of a cationic dodecapeptide (CL(14-25)), a partial sequence of cyanate lyase from rice.

Science.gov (United States)

Taniguchi, Masayuki; Takahashi, Nobuteru; Takayanagi, Tomohiro; Ikeda, Atsuo; Ishiyama, Yohei; Saitoh, Eiichi; Kato, Tetsuo; Ochiai, Akihito; Tanaka, Takaaki

2014-01-01

The antimicrobial activity of analogs obtained by substituting arginine and lysine in CL(14-25), a cationic α-helical dodecapeptide, with alanine against Porphyromonas gingivalis, a periodontal pathogen, varied significantly depending on the number and position of cationic amino acids. The alanine-substituted analogs had no hemolytic activity, even at a concentration of 1 mM. The antimicrobial activities of CL(K20A) and CL(K20A, K25A) were 3.8-fold and 9.1-fold higher, respectively, than that of CL(14-25). The antimicrobial activity of CL(R15A) was slightly lower than that of CL(14-25), suggesting that arginine at position 15 is not essential but is important for the antimicrobial activity. The experiments in which the alanine-substituted analogs bearing the replacement of arginine at position 24 and/or lysine at position 25 were used showed that arginine at position 24 was crucial for the antimicrobial activity whenever lysine at position 25 was substituted with alanine. Helical wheel projections of the alanine-substituted analogs indicate that the hydrophobicity in the vicinity of leucine at position 16 and alanines at positions 18 and/or 21 increased by substituting lysine at positions 20 and 25 with alanine, respectively. The degrees of diSC3 -5 release from P. gingivalis cells and disruption of GUVs induced by the alanine-substituted analogs with different positive charges were not closely related to their antimicrobial activities. The enhanced antimicrobial activities of the alanine-substituted analogs appear to be mainly attributable to the changes in properties such as hydrophobicity and amphipathic propensity due to alanine substitution and not to their extents of positive charge (cationicity). Copyright © 2013 Wiley Periodicals, Inc.

Alanine Enhances Aminoglycosides-Induced ROS Production as Revealed by Proteomic Analysis

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Jin-zhou Ye

2018-01-01

Full Text Available Metabolite-enabled killing of antibiotic-resistant pathogens by antibiotics is an attractive strategy to manage antibiotic resistance. Our previous study demonstrated that alanine or/and glucose increased the killing efficacy of kanamycin on antibiotic-resistant bacteria, whose action is through up-regulating TCA cycle, increasing proton motive force and enhancing antibiotic uptake. Despite the fact that alanine altered several metabolic pathways, other mechanisms could be potentially involved in alanine-mediated kanamycin killing of bacteria which remains to be explored. In the present study, we adopted proteomic approach to analyze the proteome changes induced by exogenous alanine. Our results revealed that the expression of three outer membrane proteins was altered and the deletion of nagE and fadL decreased the intracellular kanamycin concentration, implying their possible roles in mediating kanamycin transport. More importantly, the integrated analysis of proteomic and metabolomic data pointed out that alanine metabolism could connect to riboflavin metabolism that provides the source for reactive oxygen species (ROS production. Functional studies confirmed that alanine treatment together with kanamycin could promote ROS production that in turn potentiates the killing of antibiotic-resistant bacteria. Further investigation showed that alanine repressed the transcription of antioxidant-encoding genes, and alanine metabolism to riboflavin metabolism connected with riboflavin metabolism through TCA cycle, glucogenesis pathway and pentose phosphate pathway. Our results suggest a novel mechanism by which alanine facilitates kanamycin killing of antibiotic-resistant bacteria via promoting ROS production.

Volumetric property of glycine, L-serine, L-alanine and L-proline in aqueous solutions of 1-phenylpiperazinium tetrafluoroborate

International Nuclear Information System (INIS)

Xie, Hujun; Zhao, Lijiang; Liu, Chengcheng; Cao, Yifan; Lu, Xiaoxing; Lei, Qunfang; Fang, Wenjun

2016-01-01

Highlights: • Volumetric property of four amino acids in aqueous solutions of [Phpi][BF_4] were measured. • The standard partial molar volume, transparent partial molar volume and hydration number were calculated. • [Phpi][BF_4] interacts strongly with four kinds of amino acids. • Hydrophilic–hydrophobic and hydrophobic–hydrophobic interactions play the dominant roles in ternary systems. • The ternary systems are generated via multiple hydrogen bond interactions. - Abstract: The densities of aqueous solutions of glycine, L-serine, L-alanine, L-proline with the ionic liquid (IL), 1-phenylpiperazinium tetrafluoroborate ([Phpi][BF_4]) at the IL concentrations of (0.025, 0.055 and 0.100) mol·kg"−"1 have been measured at the temperatures of (298.15, 303.15 and 308.15) K. On the basis of the experimental results, the apparent molar volume (V_Φ), standard partial molar volume (V_Φ"0), transfer partial molar volume (Δ_t_rV_Φ"0) and hydration number (n_H) have been calculated. The hydrophilic–hydrophilic, hydrophobic–hydrophilic and hydrophobic–hydrophobic interactions are involved in the studied systems of {[Phpi][BF_4] + amino acids + H_2O. These volumetric parameters can help to understand the mixing effects and other complex biological processes between amino acids and ionic liquid aqueous solution.

Synthesis of {sup 15}N isotope labeled alanine; Sintese da alanina enriquecida com {sup 15}N

Energy Technology Data Exchange (ETDEWEB)

Oliveira, Claudineia R. de; Bendassolli, Jose Albertino; Sant' Ana, Carlos Roberto; Tagliassachi, Romulo Barbieri; Maximo, Everaldo; Prestes, Clelber Vieira [Centro de Energia Nuclear na Agricultura (CENA), Piracicaba, SP (Brazil). Dept. de Isotopos Estaveis]. E-mail: crolivei@cena.usp.br

2005-07-01

The application of light chemical elements and their stable isotopes in biological studies have been increased over the last years. The use of {sup 15}N labeled amino acids is an important tool for elucidation of peptides structures. This paper describe a method for the synthesis of {sup 15}N isotope labeled alanine at lower costs than international ones, as well as the details of the recovery system of the nitrogen residues. In the present work an amination of {alpha}-haloacids, with the bromopropionic carboxylic acid and labeled aqua ammonia ({sup 15}NH{sub 3} aq) was carried out. In order to avoid eventually losses of {sup 15}NH{sub 3}, special cares were adopted, since the production cost is high. Although the acquisition cost of the {sup 13}N (radioactive) labeled compounds is lower, the obtained stable tracer will allow the accomplishment of important studies of the nitrogen cycling in living things, less occupational and environment hazards, and the time limitation problems in field studies. The tests took place in triplicates with NH{sub 3} (aq) being employed. With the establishment of the system for {sup 15}NH{sub 3} recovery, an average of 94 % of the ammonia employed in the synthesis process was recovered. The purity of the amino acid was state determined by TLC (Thin Layer Chromatography) and HPLC (High-Performance Liquid Chromatography) with a fluorescence detector. The Rf and the retention time of the synthesized sample were similar the sigma standard. Finally, regarding the established conditions, it was possible to obtain the alanine with a production cost about 40 % lower than the international price. (author)

Amino Acids Regulate mTORC1 by an Obligate Two-step Mechanism*

Science.gov (United States)

Dyachok, Julia; Earnest, Svetlana; Iturraran, Erica N.; Cobb, Melanie H.

2016-01-01

The mechanistic target of rapamycin complex 1 (mTORC1) coordinates cell growth with its nutritional, hormonal, energy, and stress status. Amino acids are critical regulators of mTORC1 that permit other inputs to mTORC1 activity. However, the roles of individual amino acids and their interactions in mTORC1 activation are not well understood. Here we demonstrate that activation of mTORC1 by amino acids includes two discrete and separable steps: priming and activation. Sensitizing mTORC1 activation by priming amino acids is a prerequisite for subsequent stimulation of mTORC1 by activating amino acids. Priming is achieved by a group of amino acids that includes l-asparagine, l-glutamine, l-threonine, l-arginine, l-glycine, l-proline, l-serine, l-alanine, and l-glutamic acid. The group of activating amino acids is dominated by l-leucine but also includes l-methionine, l-isoleucine, and l-valine. l-Cysteine predominantly inhibits priming but not the activating step. Priming and activating steps differ in their requirements for amino acid concentration and duration of treatment. Priming and activating amino acids use mechanisms that are distinct both from each other and from growth factor signaling. Neither step requires intact tuberous sclerosis complex of proteins to activate mTORC1. Concerted action of priming and activating amino acids is required to localize mTORC1 to lysosomes and achieve its activation. PMID:27587390

AMINO ACID COMPOSITION OF SNAKEHEAD FISH (CHANNA STRIATUS OF VARIOUS SIZES OBTAINED AT DIFFERENT TIMES OF THE YEAR

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LAY-HARN GAM

2005-01-01

Full Text Available Snakehead fish (Channa striatus or Haruan is one of the favourite fresh water fish in the Asia-Pacific countries. The fish has been traditionally used to heal wounds. The amino acid composition of wild type Haruan was analyzed in this study. The most abundant amino acid in Haruan was glutamic acid, followed by aspartic acid, lysine, arginine, leucine, alanine, valine, threonine and glycine, in a decreasing order. The Haruan caught during rainy season was found to contain higher amount of total amino acids. The essential amino acids made up 56% of its total amino acids content. Furthermore, each of the essential amino acids (except lysine was found in higher quantity compared to other types of fishes. Haruan was found significantly rich in arginine, an important constituent in the process of wound healing. The amino acid composition of Haruan indicates that the fish is an excellent source of dietary protein for human.

Efficient L-Alanine Production by a Thermo-Regulated Switch in Escherichia coli.

Science.gov (United States)

Zhou, Li; Deng, Can; Cui, Wen-Jing; Liu, Zhong-Mei; Zhou, Zhe-Min

2016-01-01

L-Alanine has important applications in food, pharmaceutical and veterinary and is used as a substrate for production of engineered thermoplastics. Microbial fermentation could reduce the production cost and promote the application of L-alanine. However, the presence of L-alanine significantly inhibit cell growth rate and cause a decrease in the ultimate L-alanine productivity. For efficient L-alanine production, a thermo-regulated genetic switch was designed to dynamically control the expression of L-alanine dehydrogenase (alaD) from Geobacillus stearothermophilus on the Escherichia coli B0016-060BC chromosome. The optimal cultivation conditions for the genetically switched alanine production using B0016-060BC were the following: an aerobic growth phase at 33 °C with a 1-h thermo-induction at 42 °C followed by an oxygen-limited phase at 42 °C. In a bioreactor experiment using the scaled-up conditions optimized in a shake flask, B0016-060BC accumulated 50.3 g biomass/100 g glucose during the aerobic growth phase and 96 g alanine/100 g glucose during the oxygen-limited phase, respectively. The L-alanine titer reached 120.8 g/l with higher overall and oxygen-limited volumetric productivities of 3.09 and 4.18 g/l h, respectively, using glucose as the sole carbon source. Efficient cell growth and L-alanine production were reached separately, by switching cultivation temperature. The results revealed the application of a thermo-regulated strategy for heterologous metabolic production and pointed to strategies for improving L-alanine production.

Corrosion inhibition of nickel in H2SO4 solution by alanine

International Nuclear Information System (INIS)

Hamed, E.; Abd El-REhim, S.S.; El-Shahat, M.F.; Shaltot, A.M.

2012-01-01

Highlights: ► Corrosion of Ni in 1 M H 2 SO 4 in the absence and the presence of alanine. ► Alanine acts as a moderate mixed type inhibitor. ► Physical adsorption of alanine and formation of protective film are on Ni surface. ► Addition of KI improves the inhibition efficiency (synergistic effect). ► EFM technique is in reasonably good agreement with the different techniques used. - Abstract: The effect of alanine, as a safe inhibitor, was studied by measuring the corrosion of Ni in aerated and stagnant 1 M H 2 SO 4 solution (pH ∼0.2). Measurements were performed under various conditions using potentiodynamic polarization, electrochemical impedance spectroscopy (EIS) and the new electrochemical frequency modulation (EFM) methods. The obtained results showed that the addition of alanine alone gives a moderate inhibition and acts as an anodic-type inhibitor. The inhibition is due to physical adsorption of alanine on the metal surface. The inhibition efficiency enhances with increasing alanine concentration and immersion time but decreases with rise in temperature. The apparent activation energy, E a , is higher in the presence than in the absence of alanine. Addition of I − ions greatly improves the inhibition efficiency of alanine. The synergistic effect is due to enhanced adsorption of alanine cations by chemisorbed I − anions on the metal surface. The results obtained from polarization, EIS and EFM techniques are in good agreement indicating that EFM method can be used successfully for monitoring corrosion rate of Ni in H 2 SO 4 solution with and without alanine.

The simultaneous biosynthesis and uptake of amino acids by Lactococcus lactis studied by C-13-labeling experiments

DEFF Research Database (Denmark)

Jensen, N.B.S.; Christensen, B.; Nielsen, Jette

2002-01-01

Uniformly C-13 labeled glucose was fed to a lactic acid bacterium growing on a defined medium supplemented with all proteinogenic amino acids except glutamate. Aspartate stemming from the protein pool and from the extracellular medium was enriched with C-13 disclosing a substantial de novo...... biosynthesis of this amino acid simultaneous to its uptake from the growth medium and a rapid exchange flux of aspartate over the cellular membrane. Phenylalanine, alanine, and threonine were also synthesized de novo in spite of their presence in the growth medium....

Preparation of α-alanine-3H by the interaction of atomic tritium heated up to 2000 K with a solid alanine target at 77 K

International Nuclear Information System (INIS)

Filatov, Eh.S.; Simonov, E.F.; Shishkov, A.V.; Mogil'nikov, V.P.

1979-01-01

Absorption of hydrogen by alanine targets, the target behaviour and the yield of α-alanine- 3 H were studied in experiments involving straight passage of H and T atoms from the sourse (2000 K) to the target (77 K) as a function of the exposure time. In the studies with 3 H 2 the radioactivity of the gas phase was decreasing more rapidly than the overall pressure of hydrogen: H 3 H accumulates more rapidly in the gas phase. Alanine decomposition products were identified. The conditions for the studies of α-alanine- 3 H are suggested

Assessing microbial utilization of free versus sorbed Alanine by using position-specific 13C labeling and 13C-PLFA analysis

Science.gov (United States)

Herschbach, Jennifer; Apostel, Carolin; Spielvogel, Sandra; Kuzyakov, Yakov; Dippold, Michaela

2016-04-01

Microbial utilization is a key transformation process of soil organic matter (SOM). Sorption of low molecular weight organic substances (LMWOS) to soil mineral surfaces blocks or delays microbial uptake and therefore mineralization of LMWOS to CO2, as well as all other biochemical transformations. We used position-specific labeling, a tool of isotope applications novel to soil science, combined with 13C-phospholipid fatty acid (PLFA) analysis, to assess microbial utilization of sorbed and non-sorbed Alanine in soil. Alanine has various functional groups enabling different sorption mechanisms via its positive charge (e.g. to clay minerals by cation exchange), as well as via its negative charge (e.g. to iron oxides by ligand exchange). To assess changes in the transformation pathways caused by sorption, we added uniformly and position-specifically 13C and 14C labeled Alanine to the Ap of a loamy Luvisol in a short-term (10 days) incubation experiment. To allow for sorption of the tracer solution to an aliquot of this soil, microbial activity was minimized in this subsample by sterilizing the soil by γ-radiation. After shaking, the remaining solutions were filtered and the non-sorbed Alanine was removed with Millipore water and then added to non-sterilized soil. For the free Alanine treatment, solutions with Alanine of similar amount and isotopic composition were prepared, added to the soil and incubated as well. The respired CO2 was trapped in NaOH and its 14C-activity was determined at increasing times intervals. Microbial utilization of Alanine's individual C positions was evaluated in distinct microbial groups classified by 13C-PLFA analysis. Sorption to soil minerals delayed respiration to CO2 and reduced initial respiration rate by 80%. Irrespective of sorption, the highest amount was respired from the carboxylic position (C-1), whereas the amino-bound (C-2) and the methylic position (C-3) were preferentially incorporated into PLFA of microorganisms due to the

Factors that influence serum hyaluronan levels in hemodialysis patients.

Science.gov (United States)

de Medina, M; Ashby, M; Diego, J; Pennell, J P; Hill, M; Schiff, E R; Perez, G O

1999-01-01

Serum hyaluronan levels are increased in dialysis patients. We evaluated several factors that influence serum hyaluronan levels in 184 patients on chronic hemodialysis (duration 2.3 +/- 2.3 [SD] years). The levels were higher than normal in the whole group and in a subgroup of 133 patients without chronic infection, liver disease, or rheumatoid arthritis (215 +/- 19 and 205 +/- 22 microg/L, respectively). There was a tendency for the levels to be higher in a subgroup of patients with hepatitis c virus (HCV) infection. There was no correlation between hyaluronan levels, alanine aminotransferase (ALT), and duration or dose of dialysis. A weak but highly significant negative correlation between serum albumin levels and serum hyaluronan and ferritin levels was seen. The data suggest that chronic inflammation may explain, at least in part, the increased hyaluronan levels found in chronic dialysis patients.

Two alanine racemase genes in Salmonella typhimurium that differ in structure and function.

OpenAIRE

Wasserman, S A; Walsh, C T; Botstein, D

1983-01-01

Mutations were isolated in a previously undescribed Salmonella typhimurium gene encoding an alanine racemase essential for utilization of L-alanine as a source of carbon, energy, and nitrogen. This new locus, designated dadB, lies within one kilobase of the D-alanine dehydrogenase locus (dadA), which is also required for alanine catabolism. The dadA and dadB genes are coregulated. Mutants (including insertions) lacking the dadB alanine racemase do not require D-alanine for growth unless a mut...

EPR of gamma-irradiated polycrystalline alanine-in-glass dosimeter

International Nuclear Information System (INIS)

Al-Karmi, Anan M.; Morsy, M.A.

2008-01-01

This study attempts to overcome some of the reported discrepancies in alanine-EPR reproducibility that may be related to alanine dosimeter preparation and/or EPR spectrometer settings. The dosimeters were prepared by packing pure polycrystalline L-α-alanine directly as supplied by the manufacturer in glass tubes. This dosimeter production scheme avoids any possible contribution to the EPR signal from a binding material. The dosimeters were irradiated with gamma ray to low-dose ranges typical for medical therapy (0-20 Gy). Special attention has been paid to the study of minimum detectable dose, measurement repeatability and reproducibility, and post-irradiation stability. The dosimeter exhibited a linear dose response in the dose range from 0.1 to 20 Gy. These positive properties favor the polycrystalline alanine-in-glass tube as a radiation dosimeter

Theoretical and experimental approach on the molecular interactions of the DL-Alanine with an electrolytic environment

Science.gov (United States)

Cantero-López, Plinio; Yañez-Osses, Osvaldo; Páez-Meza, Manuel Silvestre; López, Johana E.; Páez-Hernández, Dayán; Arratia-Pérez, Ramiro

2017-11-01

The molecular interactions that promote the stability of proteins and amino acids in saline solutions is a central topic of molecular biophysics. However, a well-supported molecular picture of the phenomena has not been established yet. In this paper, we studied as model system the mix between DL-Alanine in aqueous solutions of STP (Na2S2O3·5H2O) at different temperatures, from volumetric and viscometric properties. The thermophysical properties obtained indicate the presence of a strong preferential solvation, structure-making action and a possible salt in effect. Quantum chemical calculations and molecular dynamic (MD) simulations provide a new insight to support these arguments.

Alanine scanning of the rabies virus glycoprotein antigenic site III using recombinant rabies virus: implication for post-exposure treatment.

Science.gov (United States)

Papaneri, Amy B; Wirblich, Christoph; Marissen, Wilfred E; Schnell, Matthias J

2013-12-02

The safety and availability of the human polyclonal sera that is currently utilized for post-exposure treatment (PET) of rabies virus (RABV) infection remain a concern. Recombinant monoclonal antibodies have been postulated as suitable alternatives by WHO. To this extent, CL184, the RABV human antibody combination comprising monoclonal antibodies (mAbs) CR57 and CR4098, has been developed and has delivered promising clinical data to support its use for RABV PET. For this fully human IgG1 cocktail, mAbs CR57 and CR4098 are produced in the PER.C6 human cell line and combined in equal amounts in the final product. During preclinical evaluation, CR57 was shown to bind to antigenic site I whereas CR4098 neutralization was influenced by a mutation of position 336 (N336) located within antigenic site III. Here, alanine scanning was used to analyze the influence of mutations within the potential binding site for CR4098, antigenic site III, in order to evaluate the possibility of mutated rabies viruses escaping neutralization. For this approach, twenty flanking amino acids (10 upstream and 10 downstream) of the RABV glycoprotein (G) asparagine (N336) were exchanged to alanine (or serine, if already alanine) by site-directed mutagenesis. Analysis of G expression revealed four of the twenty mutant Gs to be non-functional, as shown by their lack of cell surface expression, which is a requirement for the production of infectious RABV. Therefore, these mutants were excluded from further study. The remaining sixteen mutants were introduced in an infectious clone of RABV, and recombinant RABVs (rRABVs) were recovered and utilized for in vitro neutralization assays. All of the viruses were effectively neutralized by CR4098 as well as by CR57, indicating that single amino acid exchanges in this region does not affect the broad neutralizing capability of the CL184 mAb combination. Copyright © 2013 Elsevier Ltd. All rights reserved.

The Pseudomonas aeruginosa antimetabolite L-2-amino-4-methoxy-trans-3-butenoic acid (AMB is made from glutamate and two alanine residues via a thiotemplate-linked tripeptide precursor

Directory of Open Access Journals (Sweden)

Nelson eRojas Murcia

2015-03-01

Full Text Available The Pseudomonas aeruginosa toxin L-2-amino-4-methoxy-trans-3-butenoic acid (AMB is a non-proteinogenic amino acid which is toxic for prokaryotes and eukaryotes. Production of AMB requires a five-gene cluster encoding a putative LysE-type transporter (AmbA, two nonribosomal peptide synthetases (AmbB and AmbE, and two iron(II/α-ketoglutarate-dependent oxygenases (AmbC and AmbD. Bioinformatics analysis predicts one thiolation (T domain for AmbB and two T domains (T1 and T2 for AmbE, suggesting that AMB is generated by a processing step from a precursor tripeptide assembled on a thiotemplate. Using a combination of ATP-PPi exchange assays, aminoacylation assays, and mass spectrometry-based analysis of enzyme-bound substrates and pathway intermediates, the AmbB substrate was identified to be L-alanine (L-Ala, while the T1 and T2 domains of AmbE were loaded with L-glutamate (L-Glu and L-Ala, respectively. Loading of L-Ala at T2 of AmbE occurred only in the presence of AmbB, indicative of a trans loading mechanism. In vitro assays performed with AmbB and AmbE revealed the dipeptide L-Glu-L-Ala at T1 and the tripeptide L-Ala-L-Glu-L-Ala attached at T2. When AmbC and AmbD were included in the assay, these peptides were no longer detected. Instead, an L-Ala-AMB-L-Ala tripeptide was found at T2. These data are in agreement with a biosynthetic model in which L-Glu is converted into AMB by the action of AmbC, AmbD and tailoring domains of AmbE. The importance of the flanking L-Ala residues in the precursor tripeptide is discussed.

Formation of new heterocyclic amine mutagens by heating creatinine, alanine, threonine and glucose.

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Skog, K; Knize, M G; Felton, J S; Jägerstad, M

1992-08-01

A mixture of alanine, threonine, creatinine and glucose was heated in diethylene glycol and water (5:1, v/v) for 15 min at 200 degrees C. The mutagens formed were purified by high-performance liquid chromatography using the Ames/Salmonella mutagenic activity to guide the purification. The structures of the purified mutagens were determined using UV absorption, mass and NMR spectrometry. A new mutagenic compound with a mass number of 217 was found and its mass spectrum did not correspond to any known mutagen derived from food. This new compound accounted for 4% of the total mutagenic activity. Other mutagenic compounds were identified as MeIQx (2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline), 4,8-DiMeIQx (2-amino-3,4,8-trimethylimidazo[4,5-f]quinoxaline), and a new mutagen 4,7,8-TriMeIQx (2-amino-3,4,7,8-tetramethylimidazo[4,5-f]quinoxaline) with a mutagenic activity of 73,000 TA98 revertants per microgram. The percentage of the mutagenic activity attributable to MeIQx, 4,8-DiMeIQx and 4,7,8-TriMeIQx was 10%, 70% and 3%, respectively. The yield of MeIQx, 4,8-DiMeIQx and 4,7,8-TriMeIQx was 10, 36 and 6 nmole/mmole creatinine. The formation of TriMeIQx from natural meat components suggests that this new quinoxaline mutagen may be present in cooked foods.

Beta-alanine and dopamine in the reddish brown scales of Papilio butterflies

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Umebachi, Yoshishige; Ishizaki, Yumi

1983-01-01

(1) Reddish brown scales of the anal eye spot in the hind-wings of P. demoleus and P. machaon have been examined for β-alanine and dopamine. (2) The scales were fractionated into 70% ethanol-soluble fraction, 4% HCl-methanol-soluble fraction, and the residua l scales, and the β-alanine content of each fraction was determined. Most of the β-alanine present in the scales has been found in the residual scales. On acid hydrolysis of the residual scales, the β-alanine has been rather rapidly released, and the hydrolysate has contained a large amount of β-alanine. (3) The protein-bound brown pigment (HCl-ppt fraction), which was extracted with 1 N NaOH and precipitated by being acidified with HCl, has contained a large amount of β-alanine. In most or at least some of the β-alanine, the NH 2 -group has been proved to be free. (4) 14 C-Labelled β-alanine and 14 C-dopamine, which were injected at prepupal or pupal stage, have been incorporated in the highest degree into the residual scales. And the 14 C has been confirmed to be present in the HCl-ppt fraction. (5) All these results indicate that the pigment of the reddish brown scales contains β-alanine and dopamine. (author)

Gamma Amino Butyric Acid Attenuates Liver and Kidney Damage Associated with Insulin Alteration in γ-Irradiated and Streptozotocin-Treated Rats

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Saada, H.N.; Eltahawy, N.A.; Hammad, A.S.; Morcos, N.Y.S.

2016-01-01

Gamma aminobutyric acid (GABA) is one of the inhibitory neurotransmitters that may have the ability to relive the intensity of stress. The aim of the current study was to evaluate the role of γ-amino butyric acid (GABA) in modulating insulin disturbance associated with liver and kidney damage in γ-irradiated and streptozotocin-treated rats. Irradiation was performed by whole body exposure to 6 Gy from a Cs-137 source. Streptozotocin (STZ) was administered in a single intraperitoneal dose (60 mg/kg body weight). GABA (200 mg/Kg body weight/day) was administered daily via gavages during 3 weeks to γ-irradiated and STZ-treated-rats. The results obtained showed that γ-irradiation induced hyperglycemia, hyperinsulinaemia and insulin resistance (similar to type 2 Diabetes), while STZ-treatment produced hyperglycemia, insulin deficiency with no insulin resistance detected (similar to type 1 Diabetes). In both cases, significant increases of alanine amino transferase (ALT) and aspartate amino transferase (AST) activities, urea and creatinine levels were recorded in the serum. These changes were associated with oxidative damage to the liver and kidney tissues notified by significant decreases of superoxide dismutase (SOD ), catalase and glutathione peroxidase ( GSH-Px) activities in parallel to significant increases of malondialdehyde (MDA) and advanced oxidation protein products ( AOPP) levels. The administration of GABA to irradiated as well as STZ-treated rats regulated insulin and glucose levels, minimized oxidative stress and reduced the severity of liver and kidney damage. It could be concluded that GABA could be a useful adjunct to reduce some metabolic complications associated with insulin deficiency and insulin resistance

Crystal structures of active fully assembled substrate- and product-bound complexes of UDP-N-acetylmuramic acid:L-alanine ligase (MurC) from Haemophilus influenzae.

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Mol, Clifford D; Brooun, Alexei; Dougan, Douglas R; Hilgers, Mark T; Tari, Leslie W; Wijnands, Robert A; Knuth, Mark W; McRee, Duncan E; Swanson, Ronald V

2003-07-01

UDP-N-acetylmuramic acid:L-alanine ligase (MurC) catalyzes the addition of the first amino acid to the cytoplasmic precursor of the bacterial cell wall peptidoglycan. The crystal structures of Haemophilus influenzae MurC in complex with its substrate UDP-N-acetylmuramic acid (UNAM) and Mg(2+) and of a fully assembled MurC complex with its product UDP-N-acetylmuramoyl-L-alanine (UMA), the nonhydrolyzable ATP analogue AMPPNP, and Mn(2+) have been determined to 1.85- and 1.7-A resolution, respectively. These structures reveal a conserved, three-domain architecture with the binding sites for UNAM and ATP formed at the domain interfaces: the N-terminal domain binds the UDP portion of UNAM, and the central and C-terminal domains form the ATP-binding site, while the C-terminal domain also positions the alanine. An active enzyme structure is thus assembled at the common domain interfaces when all three substrates are bound. The MurC active site clearly shows that the gamma-phosphate of AMPPNP is positioned between two bound metal ions, one of which also binds the reactive UNAM carboxylate, and that the alanine is oriented by interactions with the positively charged side chains of two MurC arginine residues and the negatively charged alanine carboxyl group. These results indicate that significant diversity exists in binding of the UDP moiety of the substrate by MurC and the subsequent ligases in the bacterial cell wall biosynthesis pathway and that alterations in the domain packing and tertiary structure allow the Mur ligases to bind sequentially larger UNAM peptide substrates.

Evolutionary Diversification of Alanine Transaminases in Yeast: Catabolic Specialization and Biosynthetic Redundancy

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Ximena Escalera-Fanjul

2017-06-01

Full Text Available Gene duplication is one of the major evolutionary mechanisms providing raw material for the generation of genes with new or modified functions. The yeast Saccharomyces cerevisiae originated after an allopolyploidization event, which involved mating between two different ancestral yeast species. ScALT1 and ScALT2 codify proteins with 65% identity, which were proposed to be paralogous alanine transaminases. Further analysis of their physiological role showed that while ScALT1 encodes an alanine transaminase which constitutes the main pathway for alanine biosynthesis and the sole pathway for alanine catabolism, ScAlt2 does not display alanine transaminase activity and is not involved in alanine metabolism. Moreover, phylogenetic studies have suggested that ScALT1 and ScALT2 come from each one of the two parental strains which gave rise to the ancestral hybrid. The present work has been aimed to the understanding of the properties of the ancestral type Lacchancea kluyveri LkALT1 and Kluyveromyces lactis KlALT1, alanine transaminases in order to better understand the ScALT1 and ScALT2 evolutionary history. These ancestral -type species were chosen since they harbor ALT1 genes, which are related to ScALT2. Presented results show that, although LkALT1 and KlALT1 constitute ScALT1 orthologous genes, encoding alanine transaminases, both yeasts display LkAlt1 and KlAlt1 independent alanine transaminase activity and additional unidentified alanine biosynthetic and catabolic pathway(s. Furthermore, phenotypic analysis of null mutants uncovered the fact that KlAlt1 and LkAlt1 have an additional role, not related to alanine metabolism but is necessary to achieve wild type growth rate. Our study shows that the ancestral alanine transaminase function has been retained by the ScALT1 encoded enzyme, which has specialized its catabolic character, while losing the alanine independent role observed in the ancestral type enzymes. The fact that ScAlt2 conserves 64

Alanine-ESR dosimeter: application for dosimetry in industrial electron beam accelerator

International Nuclear Information System (INIS)

Murali, S.; Venkataramani, R.; Pushparaja; Sarma, K.S.S.; Natarajan, V.; Sastry, M.D.

2000-01-01

The feasibility of DL-α-alanine, as ESR dosimeter in powder form, was examined under the conditions of pulse electron accelerator used as an industrial irradiator. The investigations were carried out to examine the following aspects: (i) Alanine-ESR dose response in irradiator characteristics viz. various beam energies, beam currents, product conveying speeds, (ii) linearity of dose response of irradiated alanine signal for suitable range, (iii) dose uniformity of the irradiated samples and (iv) depth dose measurements using alanine powder dosimeters sandwiched between polyethylene layers. Experiments were carried out by varying some of the irradiator parameters at mobile mode of the conveyor (product under movement) and also at stationary mode for different EB energies and pulse rates. For estimation of EB dose, signal intensities of gamma irradiated DL--alanine powder calibrated with Fricke dosimetry have been used. Feasibility of application of alanine ESR dosimeter for low dose measurement down to 350 Gy has been experimentally established. The present studies show that under variable operating conditions of irradiator, alanine ESR dosimetry is suitable for dosimetric applications from low dose (350 Gy) to high dose (53 kgy). (author)

2-Phenyl-tetrahydropyrimidine-4(1H-ones – cyclic benzaldehyde aminals as precursors for functionalised β2-amino acids

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Markus Nahrwold

2009-09-01

Full Text Available Novel procedures have been developed to condense benzaldehyde effectively with β-amino acid amides to cyclic benzyl aminals. Double carbamate protection of the heterocycle resulted in fully protected chiral β-alanine derivatives. These serve as universal precursors for the asymmetric synthesis of functionalised β2-amino acids containing acid-labile protected side chains. Diastereoselective alkylation of the tetrahydropyrimidinone is followed by a chemoselective two step degradation of the heterocycle to release the free β2-amino acid. In the course of this study, an L-asparagine derivative was condensed with benzaldehyde and subsequently converted to orthogonally protected (R-β2-homoaspartate.

Study of Stationary Phase Metabolism Via Isotopomer Analysis of Amino Acids from an Isolated Protein

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Shaikh, AfshanS.; Tang, YinjieJ.; Mukhopadhyay, Aindrila; Martin, Hector Garcia; Gin, Jennifer; Benke, Peter; Keasling, Jay D.

2009-09-14

Microbial production of many commercially important secondary metabolites occurs during stationary phase, and methods to measure metabolic flux during this growth phase would be valuable. Metabolic flux analysis is often based on isotopomer information from proteinogenic amino acids. As such, flux analysis primarily reflects the metabolism pertinent to the growth phase during which most proteins are synthesized. To investigate central metabolism and amino acids synthesis activity during stationary phase, addition of fully 13C-labeled glucose followed by induction of green fluorescent protein (GFP) expression during stationary phase was used. Our results indicate that Escherichia coli was able to produce new proteins (i.e., GFP) in the stationary phase, and the amino acids in GFP were mostly from degraded proteins synthesized during the exponential growth phase. Among amino acid biosynthetic pathways, only those for serine, alanine, glutamate/glutamine, and aspartate/asparagine had significant activity during the stationary phase.

Effect of amino acids on the repression of alkaline protease synthesis in haloalkaliphilic Nocardiopsis dassonvillei

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Amit K. Sharma

2016-12-01

Full Text Available A newly isolated salt-tolerant alkaliphilic actinomycete, Nocardiopsis dassonvillei strain OK-18 grows on mineral salts medium with glucose as carbon source. It also grows and produces protease with amino acids as sole carbon source. The synthesis of extracellular alkaline protease parallel to growth was repressible by substrate concentrations. The absolute production of the protease was delinked with growth under nutritional stress, as protease production was high, despite poor growth. When amino acids served as the sole source of carbon and nitrogen, the enzyme production was significantly controlled by the number of amino acids. Maximal protease production was achieved with proline, asparagine, tyrosine, alanine, methionine and valine as sole source of carbon and nitrogen in minimal medium. With the increasing number of different amino acids in the presence and absence of glucose, the protease production was synergistically lower as compared to complex medium.

Dosimetry of electron and gamma radiation with DL-alanine

International Nuclear Information System (INIS)

Costa, Z.M. da; Campos, L.L.

1996-01-01

A dosimetric method based on the quantitative determination of stabilised free radicals in irradiated crystalline DL-alanine by electron spin resonance (ESR) spectroscopy was proposed as early in 1962. Since then, alanine dosemeters owing to their unique properties have been investigated by many authors and used in dosimetry of various types of radiation, namely gamma rays, electron and neutrons. Alanine is a simple aminoacid, on irradiation at room temperature predominantly free paramagnetic radicals of the type CH 3 -CH-COOH are produced. This paper reports the application of powder DL-alanine/ESR dosemeter for measurement of absorbed dose of gamma radiation from 60 Co sources and reactor nucleus and electron beams from accelerator. The obtained results give useful information about the instrumental care necessary to obtain the needed overall accuracy in determination of absorbed dose. (author)

Alteration in plasma free amino acid levels and its association with gout.

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Mahbub, M H; Yamaguchi, Natsu; Takahashi, Hidekazu; Hase, Ryosuke; Amano, Hiroki; Kobayashi-Miura, Mikiko; Kanda, Hideyuki; Fujita, Yasuyuki; Yamamoto, Hiroshi; Yamamoto, Mai; Kikuchi, Shinya; Ikeda, Atsuko; Kageyama, Naoko; Nakamura, Mina; Ishimaru, Yasutaka; Sunagawa, Hiroshi; Tanabe, Tsuyoshi

2017-03-16

Studies on the association of plasma-free amino acids with gout are very limited and produced conflicting results. Therefore, we sought to explore and characterize the plasma-free amino acid (PFAA) profile in patients with gout and evaluate its association with the latter. Data from a total of 819 subjects (including 34 patients with gout) undergoing an annual health examination program in Shimane, Japan were considered for this study. Venous blood samples were collected from the subjects and concentrations of 19 plasma amino acids were determined by high-performance liquid chromatography-electrospray ionization-mass spectrometry. Student's t-test was applied for comparison of variables between patient and control groups. The relationships between the presence or absence of gout and individual amino acids were investigated by logistic regression analysis controlling for the effects of potential demographic confounders. Among 19 amino acids, the levels of 10 amino acids (alanine, glycine, isoleucine, leucine, methionine, phenylalanine, proline, serine, tryptophan, valine) differed significantly (P gout. The observed significant changes in PFAA profiles may have important implications for improving our understanding of pathophysiology, diagnosis and prevention of gout. The findings of this study need further confirmation in future large-scale studies involving a larger number of patients with gout.

Bidirectional Interaction of Alanine with Sulfuric Acid in the Presence of Water and the Atmospheric Implication.

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Wang, Chun-Yu; Ma, Yan; Chen, Jiao; Jiang, Shuai; Liu, Yi-Rong; Wen, Hui; Feng, Ya-Juan; Hong, Yu; Huang, Teng; Huang, Wei

2016-04-21

Amino acids are recognized as important components of atmospheric aerosols, which impact on the Earth's climate directly and indirectly. However, much remains unknown about the initial events of nucleation. In this work, the interaction of alanine [NH2CH(CH3)COOH or Ala], one of the most abundant amino acids in the atmosphere, with sulfuric acid (SA) and water (W) has been investigated at the M06-2X/6-311++G(3df, 3pd) level of theory. We have studied thermodynamics of the hydrated (Ala)(SA) core system with up to four water molecules. We found that Ala, with one amino group and one carboxyl group, can interact with H2SO4 and H2O in two directions and that it has a high cluster stabilizing effect similar to that of ammonia, which is one of the key nucleation precursor. The corresponding Gibbs free energies of the (Ala)(SA)(W)n (n = 0-4) clusters formation at 298.15 K predicted that Ala can contribute to the stabilization of small binary clusters. Our results showed that the hydrate distribution is temperature-dependent and that a higher humidity and temperature can contribute to the formation of hydrated clusters.

Effects of preservation methods on amino acids and 5'-nucleotides of Agaricus bisporus mushrooms.

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Liu, Ying; Huang, Fan; Yang, Hong; Ibrahim, S A; Wang, Yan-Feng; Huang, Wen

2014-04-15

In this study, the proximate composition, free amino acids content and 5'-nucleotides in frozen, canned and salted Agaricus bisporus (A. bisporus) were investigated. We found that the three kinds of A. bisporus products were good sources of protein, with amount varying in the ranges of 16.54-24.35g/100g (dry weight). Freezing, canning and salting process, followed by 6months of storage led to a significant reduction in free amino acids, especially tyrosine, alanine, glutamine and cysteine. There were medium levels of MSG-like amino acids in frozen A. bisporus and canned A. bisporus, and low levels of MSG-like amino acids in salted A. bisporus. The mount of flavor 5'-nucleotides in frozen A. bisporus was higher than that of canned and salted A. bisporus. The present study thus suggests that freezing is beneficial for the preservation of A. bisporus. Copyright © 2013 Elsevier Ltd. All rights reserved.

Gross and true ileal digestible amino acid contents of several animal body proteins and their hydrolysates.

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Cui, J; Chong, B; Rutherfurd, S M; Wilkinson, B; Singh, H; Moughan, P J

2013-07-01

Amino acid compositions of ovine muscle, ovine myofibrillar protein, ovine spleen, ovine liver, bovine blood plasma, bovine blood globulins and bovine serum albumin and the amino acid compositions and in vivo (laboratory rat) true ileal amino acid digestibilities of hydrolysates (sequential hydrolysis with Neutrase, Alcalase and Flavourzyme) of these protein sources were determined. True ileal amino acid digestibility differed (Pprotein hydrolysates. The ovine myofibrillar protein and liver hydrolysates were the most digestible, with a mean true ileal digestibility across all amino acids of 99%. The least digestible protein hydrolysate was bovine serum albumin with a comparable mean true ileal digestibility of 93%. When the digestible amino acid contents were expressed as proportions relative to lysine, considerable differences, across the diverse protein sources, were found in the pattern of predicted absorbed amino acids. Copyright © 2013 Elsevier Ltd. All rights reserved.

Effect of amino acids on red pigments and citrinin production in Monascus ruber.

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Hajjaj, Hassan; François, Jean-Marie; Goma, Gérard; Blanc, Philippe J

2012-03-01

Amino acids were used as sole nitrogen sources to examine their effects on the production of water-soluble red pigments and citrinin by Monascus ruber ATCC 96218 cultivated on chemically defined media. In general, when glycine, tyrosine, arginine, serine, or histidine were used as sole nitrogen sources, they favored the production of red pigments, and restricted the synthesis of the mycotoxin. In contrast, the production of citrinin was enhanced in media supplemented with either glutamate, alanine, or proline. Histidine was found to be the most valuable amino acid as it resulted in the highest production of red pigments and almost completely eliminated the formation of mycotoxin. © 2012 Institute of Food Technologists®

Photolabile protection for amino acids: studies on the release from novel benzoquinolone cages.

Science.gov (United States)

Fonseca, Andrea S C; Soares, Ana M S; Gonçalves, M Sameiro T; Costa, Susana P G

2015-12-01

The synthesis of a novel fused nitrogen heterocycle, benzoquinolone, for evaluation as a photocleavable protecting group is described for the first time by coupling to model amino acids (alanine, phenylalanine and glutamic acid). Conversion of the phenylalanine ester conjugate to the thionated derivative was accomplished by reaction with Lawesson's reagent. Photocleavage studies of the carbonyl and thiocarbonyl benzoquinolone conjugates in various solvents and at different wavelengths (300, 350 and 419 nm) showed that the most interesting result was obtained at 419 nm for the thioconjugate, revealing that the presence of the thiocarbonyl group clearly improved the photolysis rates, giving practicable irradiations times for the release of the amino acids (less than 1 min).

Status report of the ESR/alanine project of the IAEA Dosimetry Laboratory

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