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Sample records for alanine transaminase

  1. 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 ...

  2. beta-Chloro-L-alanine inhibition of the Escherichia coli alanine-valine transaminase.

    OpenAIRE

    Whalen, W A; Wang, M D; Berg, C M

    1985-01-01

    beta-Chloro-L-alanine, an amino acid analog which inhibits a number of enzymes, reversibly inhibited the Escherichia coli K-12 alanine-valine transaminase, transaminase C. This inhibition, along with the inhibition of transaminase B, accounted for the isoleucine-plus-valine requirement of E. coli in the presence of beta-chloro-L-alanine.

  3. Porcine alanine transaminase after liver allo-and xenotransplantation

    OpenAIRE

    Ekser, Burcin; Gridelli, Bruno; Cooper, David K.C.

    2012-01-01

    Aspartate transaminase (AST) and alanine transaminase (ALT) are measured following liver transplantation as indicators of hepatocellular injury. During a series of orthotopic liver allo-and xenotransplants, we observed that there was an increase in AST in all cases. The anticipated concomitant rise in ALT did not occur when a wild-type (WT) pig was the source of the liver graft, but did occur when a baboon or a genetically engineered (α1,3-galactosyltransferase gene-knockout [GTKO]) pig was t...

  4. Porcine alanine transaminase after liver allo-and xenotransplantation.

    Science.gov (United States)

    Ekser, Burcin; Gridelli, Bruno; Cooper, David K C

    2012-01-01

    Aspartate transaminase (AST) and alanine transaminase (ALT) are measured following liver transplantation as indicators of hepatocellular injury. During a series of orthotopic liver allo-and xenotransplants, we observed that there was an increase in AST in all cases. The anticipated concomitant rise in ALT did not occur when a wild-type (WT) pig was the source of the liver graft, but did occur when a baboon or a genetically engineered (α1,3-galactosyltransferase gene-knockout [GTKO]) pig was the source of the graft. We hypothesized that the cience of Galα1,3Gal in GTKO pig livers may render pig hepatocytes similar to human and baboon hepatocytes in their response to hepatocellular injury. Reviewing the literature, after WT pig liver allotransplantation or xenotransplantation, in the majority of reports, although changes in AST were reported, no mention was made of changes in ALT, suggesting that there was no change in ALT. However, Ramirez et al. reported two cases of liver xenotransplants from hCD55 pigs, following which there were increases in both AST and ALT, suggesting that it is not simply the cience of expression of Galα1,3Gal that is the cause. We acknowledge that our observation is based on a small number of experiments, but we believe it is worth recording. © 2012 John Wiley & Sons A/S.

  5. 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

  6. and alanine (EC. 2.6.1.2) transaminases, and alkaline phosphatase

    African Journals Online (AJOL)

    The activities of aspartate (E.C. 2.6.1.1) and alanine (E.C. 2.6.1.2) transaminases (AST and ALT, respectively), and alkaline phosphatase (ALP) (E.C. 3.1.3.1) were determined in erythrocytes obtained from 20 HbAA, 15 HbAS and 12 HbSS human subjects. The results showed that the three enzymes had different levels of ...

  7. Occult hepatitis B virus among the patients with abnormal alanine transaminase.

    Science.gov (United States)

    Makvandi, Manoochehr; Neisi, Niloofar; Khalafkhany, Davod; Makvandi, Kamyar; Hajiani, Eskandar; Shayesteh, Ali Akbar; Masjedi Zadeh, Abdolrahim; Sina, Amir Hosein; Hamidifard, Mojtaba; Rasti, Mojtaba; Aryan, Ehsan; Ahmadi, Kambiz; Yad Yad, Mohammad Jafar

    2014-08-01

    The occult hepatitis B infection (OBI) is defined as the presence of hepatitis B virus (HBV) DNA in the sera or in the liver biopsy and the absence of hepatitis B surface antigen (HBsAg) by serological test. The current study aimed to evaluate the occult HBV infection by polymerase chain reaction (PCR) and determine HBV genotyping among the patients with abnormal alanine transaminase (ALT) in Ahvaz city, Iran. The sera of 120 patients, 54 (45%) females and 66 (55%) males, with abnormal ALT 40-152 IU were collected. All the patients were negative for HBsAg for more than one year. The patients` sera were tested by PCR using primers specified for the S region of HBV. Then the positive PCR products were sequenced to determine HBV genotyping and phylogenic tree. Of these 120 subjects, 12 (10%) patients including 6 (5%) males and 6 (5%) females were found positive for HBV DNA by PCR, which indicated the presence of occult HBV infection among these patients. The sequencing results revealed that genotype D was predominant with sub-genotyping D1 among OBI patients. Occult hepatitis B infection is remarkably prevalent in Ahvaz, Iran, and should be considered as a potential risk factor for the transmission of Hepatitis B Virus throughout the community by the carriers.

  8. The predictive value of hospital admission serum alanine transaminase activity in patients treated for paracetamol overdose.

    Science.gov (United States)

    Al-Hourani, K; Mansi, R; Pettie, J; Dow, M; Bateman, D N; Dear, J W

    2013-06-01

    Paracetamol is a major cause of poisoning. Treatment decisions are predominately based on the dose ingested and a timed blood paracetamol concentration because most patients present to hospital soon after overdose, before hepatotoxicity can be confirmed/excluded using serum alanine transaminase (ALT). Nonetheless, ALT is measured at hospital presentation; we investigated its value in predicting hepatotoxicity. From March 2011 to May 2012, patients admitted to the Royal Infirmary of Edinburgh for paracetamol overdose treatment were identified. We determined the value of admission ALT (below or above our upper limit of normal-50 IU/l) at predicting three endpoints: 1-doubling of ALT; 2-peak ALT >1000 IU/l; 3-peak international normalized ratio (INR) >2. From 500 patients, 410 met the entry criteria; 264 presented within 8 h of overdose, 54 between 8 and 24 h, 53 after 24 h and 39 were staggered ingestions. Admission ALT was increased in 71. For endpoint 1 (ALT doubling), the positive predictive value (PPV) of admission ALT was 19% [95% confidence interval (CI) 12-30] with a negative predictive value (NPV) of 98% (95% CI 96-99); endpoint 2 (ALT >1000 IU/l: PPV 23% (95% CI 14-34) and NPV 100% (95% CI 99-100) and for endpoint 3 (INR >2): PPV 14% (95% CI 7-25) and NPV of 100% (95% CI 99-100). The NPV remained high when only late presenters were included. Admission ALT within the normal range has a high NPV and could be used, alone or in combination with newer biomarkers, to identify lower risk patients at hospital presentation.

  9. AKTIVITAS ASPARTAT AMINOTRANSFERASE (AST DAN ALANIN TRANSAMINASE (ALT PADA MONYET EKOR PANJANG (Macaca fascicularis OBESITAS DI PURA LUHUR ULUWATU, BALI

    Directory of Open Access Journals (Sweden)

    Ayu Paramita Lestari

    2016-08-01

    Full Text Available Penelitian obsevasional-deskriptif dengan pendekatan cross-sectional telah dilakukan untuk mengetahui aktivitas aspartat aminotransferase (AST dan alanin transaminase (ALT pada monyet ekor panjang (Macaca fascicularis obesitas yang hidup liar di kawasan Pura Luhur Uluwatu, Bali. Sebanyak 16 ekor monyet ekor panjang berhasil dibius menggunakan ketamine dosis 10 mg/kg berat badan dicampur dengan premedikasi xylasin dosis 1-2 mg/kg berat badan. Sampel yang digunakan adalah serum darah monyet obesitas yang diambil dari monyet dalam keadaan terbius. Dari 16 ekor monyet, 12 ekor tergolong obesitas berdasarkan Indeks Massa Tubuh dan Berat Badan. Aktivitas aspartat aminotransferase (AST dan alanin transaminase (ALT ditentukan menggunakan mesin automatic chemistry analyzer (Indiko-Thermo Scientific. Hasil penelitian menunjukkan nilai AST bervariasi dari 43-88 U/L dan ALT 26-77 U/L dengan rataan 69,4 ± 14,9 U/L. Dari hasil penelitian dapat disimpulkan bahwa nilai AST dan ALT monyet obesitas cenderung lebih tinggi dari nilai AST dan ALT normal.

  10. Effect of 28-Day Exposure to Fenpropathrin on the Activities of Serum Alanine Transaminase and Liver Antioxidant Enzymes in Mice

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    Nieradko-Iwanicka Barbara

    2015-04-01

    Full Text Available Fenpropathrin (FEN was administered intraperitoneally in doses of 2.38 mg/kg, 5.9 5mg/kg, or 11.9 mg/kg b.w. to mice for 28 consecutive days. FEN did not significantly affect the activity of alanine transaminase (ALT in the sera. Superoxide dismutase and glutathione peroxidase activities were significantly elevated in the liver after a 28-d exposure to moderate or highest doses of the pesticide. These results demonstrate that the 28-day exposure to FEN leads to an up-regulation of expression of antioxidant enzymes in response to an oxidative stress in mouse liver without causing a significant increase in ALT activity.

  11. Detection of Hepatitis C virus RNA in peripheral blood mononuclear cells of patients with abnormal alanine transaminase in Ahvaz

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    M Makvandi

    2014-01-01

    Full Text Available Purpose: Hepatitis C virus (HCV is an important agent for chronic and acute hepatitis. Occult hepatitis C remains a major health problem worldwide. Patients with chronic occult HCV may progress to cirrhosis and hepatocellular carcinoma. The aim of this study was to determine prevalence of occult hepatitis C by IS-PCR-ISH (in situ PCR in situ hybridisation in the patients with abnormal ALT. Materials and Methods: The blood samples were taken from 53 patients including 17 females (32.1% and 36 (67.9% males who had abnormal alanine transaminase (ALT for more than 1 year. The mean ALT and aspartate transaminase (AST level were 41.02 ± 9.3 and 24.17 ± 7.3, respectively. The patients′ age were between 4 and 70-years old with mean age 38 ± 13. All the patients were negative for HCV antibody, HCV RNA and HBs Ag. The peripheral blood mononuclear cells (PBMC were separated with ficoll gradient from each blood sample, then the cells were fixed on slides by cold acetone and followed by IS-PCR-ISH for HCV RNA detection. Results: Seventeen (32% patients including 6 (11.3% females and 11 (20.7% males showed positive results for HCV RNA by in situ-PCR in situ hybridisation. Ten (18.8% positive cases were between 20 and 40-years old and 6 (11.3% positive patients were between 40 and 60 years old. Ten (19.6% patients who were positive for IS-PCR-ISH also had positive anti-HBc IgG and 7 (13.2% patients were negative for HBc-IgG. Conclusion: In the present study high rate of 32% occult hepatitis C were found among the patients with elevated ALT.

  12. The effectiveness of fermented turmeric powder in subjects with elevated alanine transaminase levels: a randomised controlled study

    Science.gov (United States)

    2013-01-01

    Background Previous animal studies have shown that Curcuma longa (turmeric) improves liver function. Turmeric may thus be a promising ingredient in functional foods aimed at improving liver function. The purpose of the study is to investigate the hepatoprotective effect of fermented turmeric powder (FTP) on liver function in subjects with elevated alanine transaminase (ALT) levels. Methods A randomised, double-blind, placebo-controlled trial was conducted between November 2010 and April 2012 at the clinical trial center for functional foods of the Chonbuk National University Hospital. The trial included 60 subjects, 20 years old and above, who were diagnosed mild to moderate elevated ALT levels between 40 IU/L and 200 IU/L. Sixty subjects were randomised to receive FTP 3.0 g per day or placebo 3.0 g per day for 12 weeks. The treatment group received two capsules of FTP three times a day after meals, for 12 weeks. The primary efficacy endpoint was change in the ALT levels in the two groups. The secondary efficacy endpoints included its effect on aspartate aminotransferase (AST), gamma-glutamyl transferase (GGT), total bilirubin (TB), and lipid profiles. Safety was assessed throughout the study using ongoing laboratory tests. Adverse events (AEs) were also recorded. Results Sixty subjects were randomised in the study (30 into the FTP group, 30 into the placebo group), and among them, twelve subjects were excluded from the analysis for protocol violation, adverse events or consent withdrawal. The two groups did not differ in baseline characteristics. After 12 weeks of treatment, 48 subjects were evaluated. Of the 48 subjects, 26 randomly received FTP capsules and 22 received placebo. The FTP group showed a significant reduction in ALT levels after 12 weeks of treatment compared with the placebo group (p = 0.019). There was also observed that the serum AST levels were significantly reduce in the FTP group than placebo group (p = 0.02). The GGT levels

  13. Evaluation of Preoperative Aspartate Transaminase/Alanine Transaminase Ratio as an Independent Predictive Biomarker in Patients With Metastatic Renal Cell Carcinoma Undergoing Cytoreductive Nephrectomy: A Propensity Score Matching Study.

    Science.gov (United States)

    Ishihara, Hiroki; Kondo, Tsunenori; Yoshida, Kazuhiko; Omae, Kenji; Takagi, Toshio; Iizuka, Junpei; Tanabe, Kazunari

    2017-10-01

    The usefulness of the aspartate transaminase (AST)/alanine transaminase (ALT) ratio (De Ritis ratio) as a predictive biomarker for patients with metastatic renal cell carcinoma (mRCC) undergoing cytoreductive nephrectomy (CN) remains unclear. The data from 118 patients were retrospectively evaluated. The endpoints were cancer-specific survival (CSS) and overall survival (OS) after CN. We compared these according to the AST/ALT ratio before and after 1:1 propensity score matching. The independent predictors for CSS and OS were also analyzed. The area under the receiver operating characteristic curve was 0.603. The maximum Youden index indicated that the cutoff value for the AST/ALT ratio was 1.24. Before matching, a high AST/ALT ratio was significantly associated with inferior CSS and OS (P AST/ALT ratio groups. In the matched cohort, CSS and OS tended to be lower in the high AST/ALT ratio group, although the results were not statistically significant (median CSS, 18.4 months vs. not reached, P = .121; OS, 18.4 months vs. not reached, P = .0957). Furthermore, multivariate analyses revealed that the AST/ALT ratio was an independent predictor for CSS and OS (CSS hazard ratio, 2.17, P = .0472; OS hazard ratio, 2.30, P = .0258). The preoperative AST/ALT ratio can be an effective predictive biomarker for CSS and OS in patients with mRCC. Copyright © 2017 Elsevier Inc. All rights reserved.

  14. 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.

  15. Correlations of Complete Blood Count with Alanine and Aspartate Transaminase in Chinese Subjects and Prediction Based on Back-Propagation Artificial Neural Network (BP-ANN).

    Science.gov (United States)

    Yu, Jiong; Pan, Qiaoling; Yang, Jinfeng; Zhu, Chengxing; Jin, Linfeng; Hao, Guangshu; Shi, Xiaowei; Cao, Hongcui; Lin, Feiyan

    2017-06-19

    BACKGROUND The complete blood count (CBC) is the most common examination used to monitor overall health in clinical practice. Whether there is a relationship between CBC indexes and alanine transaminase (ALT) and aspartate aminotransferase (AST) has been unclear. MATERIAL AND METHODS In this study, 572 normal-weight and 346 overweight Chinese subjects were recruited. The relationship between CBC indexes with ALT and AST were analyzed by Pearson and Spearman correlations according to their sex, then we conducted colinearity diagnostics and multiple linear regression (MLR) analysis. A prediction model was developed by a back-propagation artificial neural network (BP-ANN). RESULTS ALT was related to 4 CBC indexes in the male normal-weight group and 3 CBC indexes in the female group. In the overweight group, ALT had a similar relationship with the normal group, but there was only 1 index related with AST in the normal-weight group and male overweight groups. The ALT regression models were developed in normal-weight and overweight people, which had better correlation coefficient (R>0.3). After training 1000 epochs, the BP-ANN models of ALT achieved higher correlations than MLR models in normal-weight and overweight people. CONCLUSIONS ALT is a more suitable index than AST for developing a regression model. ALT can be predicted by CBC indexes in normal-weight and overweight individuals based on a BP-ANN model, which was better than MLR analysis.

  16. Factors Predicting HBsAg Seroclearance and Alanine Transaminase Elevation in HBeAg-Negative Hepatitis B Virus-Infected Patients with Persistently Normal Liver Function.

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    Tai-Long Chien

    Full Text Available A certain proportion of hepatitis B virus (HBV-infected patients with persistently normal alanine transaminase (ALT levels have significant fibrosis. Using liver stiffness measurements (Fibroscan® and laboratory data, including serum ALT, quantitative HBsAg (qHBsAg, and HBV DNA, we attempted to predict the natural histories of these patients.Non-cirrhotic HBeAg-negative chronic hepatitis B patients with persistently normal ALT were followed up prospectively with the end points of HBsAg seroclearance and ALT elevation above the upper limit of normal. The factors that were predictive of the end points were identified.A total of 235 patients with an average age of 48.1 +/- 10.7 years were followed up for 7 years. Eight patients (3.4% lost HBsAg, and 15 patients (6.4% experienced ALT elevation. The overall cumulative HBsAg seroclearances were 0.4%, 1.3% and 2.3% at years 1, 3 and 5, respectively. Regarding HBsAg seroclearance, the qHBsAg (< 30 IU/ml cutoff resulted in a hazard ratio (HR of 19.6 with a 95% confidence interval (CI of 2.2-166.7 (P = 0.008. The baseline ALT level (odd ratio (OR 1.075, 95% CI 1.020-1.132, P = 0.006 and a qHBsAg above 1000 IU/ml (3.7, 1.1-12.4, P = 0.032 were associated with ALT elevation. Limited to men, the baseline liver stiffness (1.6, 1.0-2.5, P = 0.031 and a qHBsAg above 1000 IU/ml (10.4, 2.1-52.4, P = 0.004 were factors that were independently associated with ALT elevation.A low qHBsAg level predicted HBsAg clearance. Baseline ALT and a qHBsAg above 1000 IU/ml were independent predictive factors for ALT elevation. Among the men, the independent predictive factors for ALT elevation were qHBsAg and liver stiffness.

  17. EFFECT OF CIGARETTE SMOKING ON SERUM TRANSAMINASES ...

    African Journals Online (AJOL)

    EFFECT OF CIGARETTE SMOKING ON SERUM TRANSAMINASES. ... Global Journal of Medical Sciences ... Alanine transaminase and aspartate transaminase activity were assessed in 20 mild to moderate smokers, 20 heavy smokers and 20 non smokers, all males, aged between 17 and 35 years in Calabar Cross River ...

  18. Incidence and predictive factors of transaminase elevation in patients consulting for dengue fever in Cayenne Hospital, French Guiana.

    Science.gov (United States)

    Djossou, Félix; Vesin, Guillaume; Walter, Gaelle; Epelboin, Loïc; Mosnier, Emilie; Bidaud, Bastien; Abboud, Philippe; Okandze, Antoine; Mattheus, Severine; Elenga, Narcisse; Demar, Magalie; Malvy, Denis; Nacher, Mathieu

    2016-02-01

    The objective of the study was to determine the incidence of transaminase elevation during dengue, and its predictive factors. In 2013, a longitudinal study was performed using data from all cases of dengue seen in Cayenne Hospital. Cox proportional modeling was used. Signs of major transaminase elevation were defined as an increase in aspartate amino transferase (AST) or alanine amino transferase (ALT) concentration over 10 times the normal value (10N). There were 1574 patients and 13 249 person-days of follow-up. The incidence rate for signs of transaminase elevation (10N) was 0.55 per 100 person-days. Six patients had major transaminase elevation with AST>1000 units (0.43 per 1000 patient-days), and 73 patients (4.6%) developed transaminase elevation with AST >10N. The variables independently associated with major transaminase elevation were hyponatremia, low platelets, dehydration, hematocrit increase, food intolerance, positive nonstructural protein 1 (NS1), age over 15 years and the notion of paracetamol intake. Although very frequent, the incidence of major transaminase elevation was lower than reported elsewhere perhaps because of good access to care, or of the particular serotype causing this epidemic. The patients with transaminase elevation tended to be older, more severe and taking paracetamol. . © The Author 2016. Published by Oxford University Press on behalf of Royal Society of Tropical Medicine and Hygiene. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  19. Glutamic Pyruvate Transaminase GPT2 Promotes Tumorigenesis of Breast Cancer Cells by Activating Sonic Hedgehog Signaling

    OpenAIRE

    Cao, Yuan; Lin, Shu-Hai; Wang, Yongbin; Chin, Y. Eugene; Kang, Lan; Mi, Jun

    2017-01-01

    Increased glutamine metabolism is a hallmark of cancer. Mitochondrial glutamic pyruvate transaminase (GPT2) catalyzes the reversible transamination between alanine and ?-ketoglutarate (?-KG), also known as 2-oxoglutarate, to generate pyruvate and glutamate during cellular glutamine catabolism. However, the precise role of GPT2 in tumorigenesis remains elusive. Here, we report that in breast cancer tissue samples and breast cancer cell lines, GPT2 expression level was markedly elevated and cor...

  20. The effect of antioxidant supplementation on hepatitis C viral load, transaminases and oxidative status: a randomized trial among chronic hepatitis C virus-infected patients

    DEFF Research Database (Denmark)

    Groenbaek, K.; Friis, H.; Hansen, Max

    2006-01-01

    Objective To assess the effect of antioxidant supplementation on hepatitis C viral load, transaminases and oxidative status. Methods We performed a randomized, placebo-controlled, double-blind trial to assess the effect of antioxidant supplementation on serum alanine aminotransferase, plasma hepa...... and catalase) or plasma levels of oxidative markers (malondialdehyde and 2-amino-adipic semialdehyde) were found. Conclusion Supplementation with vitamin C, E and selenium increased the antioxidant status, but had no effects on alanine aminotransferase, viral load or oxidative markers....

  1. A Process Concept for High-Purity Production of Amines by Transaminase-Catalyzed Asymmetric Synthesis: Combining Enzyme Cascade and Membrane-Assisted ISPR

    DEFF Research Database (Denmark)

    Börner, Tim; Rehn, Gustav; Grey, Carl

    2015-01-01

    For the amine transaminase (ATA)-catalyzed synthesis of chiral amines, the choice of donor substrate is of high importance for reaction and process design. Alanine was investigated as an amine donor for the reductive amination of a poorly water-soluble ketone (4-phenyl-2-butanone) in a combined...

  2. Alanine water complexes.

    Science.gov (United States)

    Vaquero, Vanesa; Sanz, M Eugenia; Peña, Isabel; Mata, Santiago; Cabezas, Carlos; López, Juan C; Alonso, José L

    2014-04-10

    Two complexes of alanine with water, alanine-(H2O)n (n = 1,2), have been generated by laser ablation of the amino acid in a supersonic jet containing water vapor and characterized using Fourier transform microwave spectroscopy. In the observed complexes, water molecules bind to the carboxylic group of alanine acting as both proton donors and acceptors. In alanine-H2O, the water molecule establishes two intermolecular hydrogen bonds forming a six-membered cycle, while in alanine-(H2O)2 the two water molecules establish three hydrogen bonds forming an eight-membered ring. In both complexes, the amino acid moiety is in its neutral form and shows the conformation observed to be the most stable for the bare molecule. The microsolvation study of alanine-(H2O)n (n = 1,2) can be taken as a first step toward understanding bulk properties at a microscopic level.

  3. Reference values for alanine and aspartate aminotransferase (ALT and AST) optimized by addition of pyridoxal phosphate.

    Science.gov (United States)

    Plebani, M; Bonvicini, P; De Besi, T; Giordano, A; Pesarin, F; Ceriotti, G

    1980-01-01

    The increases of the activity of aspartate and alanine transaminases observed in sera of healthy male and female subjects by addition of pyridoxal phosphate have been determined. The reference values of the activated reactions, distinguished by sex and age classes, are presented.

  4. Liver alanine aminotransferase, insulin resistance and endothelial dysfunction in normotriglyceridaemic subjects with type 2 diabetes mellitus

    NARCIS (Netherlands)

    Schindhelm, RK; Diamant, M; Bakker, SJL; van Dijk, RAJM; Scheffer, PG; Teerlink, T; Kostense, PJ; Heine, RJ

    Background Plasma levels of liver transaminases, including alanine aminotransferase (ALT), are elevated in most cases of nonalcoholic fatty liver disease (NAFLD). Elevated ALT levels are associated with insulin resistance, and subjects with NAFLD have features of the metabolic syndrome that confer

  5. Studies on transaminase reactions in some parasitic helminths.

    Science.gov (United States)

    Min, Yong Ok; Seo, Byong Seol

    1966-12-01

    By an application of Sigma-Frankel methods, two transaminase systems, glutamic-pyruvic transaminase and glutamic-oxaloacetic transaminase, were found to operate at a mesurable rate in 2 species of nematodes(Ascaris lumbricoides and Ascaridia galli), 5 species of trematodes (Clonorchis sinensis, Fasciola hepatica, Eurytrema pancreaticum, Paramphistomum cervi and Paragonimus westermani) and 5 kinds of cestodes (Diphyllobothrium mansoni, Dipylidium caninum, Taenia pisiformis, Cysticercus cellulosae and Cysticercus pisiformis). A comparison was made of the transamination reactions in nematodes and those of trematodes and cestodes. And the significance of transaminase in these parasites is discussed in relation to protein synthesis and its utilization.

  6. Impact of antiretroviral therapy on glycemia and transaminase levels in patients living with HIV/AIDS in Limbe, Cameroon.

    Science.gov (United States)

    Ottop, Francisca M; Atashili, Julius; Ndumbe, Peter M

    2013-01-01

    This study aimed to determine the effect of antiretroviral therapy (ART) on glycemia and transaminase levels in HIV-infected patients in Limbe, Cameroon. A total of 200 ART-experienced patients and 30 ART-naive HIV-infected controls were recruited after submitting signed informed consent forms. The blood samples collected were screened for hepatitis B and C, and liver transaminases and random blood sugar (RBS) levels were determined spectrophotometrically. No significant correlation existed between the RBS and transaminase levels with either the duration of ART or the duration since HIV diagnosis. Although both groups of patients appeared to have similar transaminase and RBS levels, after controlling for confounders, patients on ART had significantly higher aspartate aminotransferase levels, significantly lower alanine aminotransferase levels, and slightly (but not significantly) lower glycemia levels (respective mean differences of 14.92 IU/L, P = .00; 11.95 IU/L, P = .00; and 7.60 mg/dL, P = .19). These changes need to be considered in monitoring patients on ART in this setting and other similar settings.

  7. Role of L-alanine for redox self-sufficient amination of alcohols.

    Science.gov (United States)

    Klatte, Stephanie; Wendisch, Volker F

    2015-01-23

    In white biotechnology biocatalysis represents a key technology for chemical functionalization of non-natural compounds. The plasmid-born overproduction of an alcohol dehydrogenase, an L-alanine-dependent transaminase and an alanine dehydrogenase allows for redox self-sufficient amination of alcohols in whole cell biotransformation. Here, conditions to optimize the whole cell biocatalyst presented in (Bioorg Med Chem 22:5578-5585, 2014), and the role of L-alanine for efficient amine functionalization of 1,10-decanediol to 1,10-diaminodecane were analyzed. The enzymes of the cascade for amine functionalization of alcohols were characterized in vitro to find optimal conditions for an efficient process. Transaminase from Chromobacterium violaceum, TaCv, showed three-fold higher catalytic efficiency than transaminase from Vibrio fluvialis, TaVf, and improved production at 37°C. At 42°C, TaCv was more active, which matched thermostable alcohol dehydrogenase and alanine dehydrogenase and improved the 1,10-diaminodecane production rate four-fold. To study the role of L-alanine in the whole cell biotransformation, the L-alanine concentration was varied and 1,10.diaminodecane formation tested with constant 10 mM 1,10- decanediol and 100 mM NH4Cl. Only 5.6% diamine product were observed without added L-alanine. L-alanine concentrations equimolar to that of the alcohol enabled for 94% product formation but higher L-alanine concentrations allowed for 100% product formation. L-alanine was consumed by the E. coli biocatalyst, presumably due to pyruvate catabolism since up to 16 mM acetate accumulated. Biotransformation employing E. coli strain YYC202/pTrc99a-ald-adh-ta Cv, which is unable to catabolize pyruvate, resulted in conversion with a selectivity of 42 mol-%. Biotransformation with E. coli strains only lacking pyruvate oxidase PoxB showed similar reduced amination of 1,10-decanediol indicating that oxidative decarboxylation of pyruvate to acetate by PoxB is primarily

  8. Effects of lixisenatide on elevated liver transaminases

    DEFF Research Database (Denmark)

    Gluud, Lise L; Knop, Filip K; Vilsbøll, Tina

    2014-01-01

    identified on AST, alkaline phosphatase or bilirubin. No evidence of bias was identified. Mixed effect multilevel meta-regression analyses suggest that the benefit of lixisenatide on ALT was limited to patients who were overweight or obese. CONCLUSIONS: This review suggests that lixisenatide increases......) on lixisenatide versus placebo or active comparators for type 2 diabetes were included. PARTICIPANTS: Individual patient data were retrieved to calculate outcomes for patients with elevated liver blood tests. MAIN OUTCOME MEASURES: Normalisation of alanine aminotransferase (ALT) and aspartate aminotransferase...... (AST). DATA SYNTHESIS: The results of included trials were combined in meta-analyses. Sequential, subgroup and regression analyses were performed to evaluate heterogeneity and bias. RESULTS: We included 12 RCTs on lixisenatide versus placebo and 3 RCTs with the active comparators liraglutide, exenatide...

  9. A case study on robust optimal experimental design for model calibration of ω-Transaminase

    DEFF Research Database (Denmark)

    Daele, Timothy, Van; Van Hauwermeiren, Daan; Ringborg, Rolf Hoffmeyer

    Proper calibration of models describing enzyme kinetics can be quite challenging. This is especially the case for more complex models like transaminase models (Shin and Kim, 1998). The latter fitted model parameters, but the confidence on the parameter estimation was not derived. Hence, the usabi......Proper calibration of models describing enzyme kinetics can be quite challenging. This is especially the case for more complex models like transaminase models (Shin and Kim, 1998). The latter fitted model parameters, but the confidence on the parameter estimation was not derived. Hence......, the usability of the parameter estimates is difficult to assess.In this paper, the confidence is derived, using the Fisher Information Matrix (FIM) for the backward reaction (conversion of acetophenone and alanine to α-methylbenzylamine and pyruvate). FIM computation requires local parameter sensitivities...... is to make the design less dependent on one specific parameter set, but make it suitable for a subset of parameters in a local parameter space.This robust OED methodology is currently being applied to the backward part of the model of Shin and Kim (1998) to design experiments for the conversion of 1-methyl-2...

  10. PEDIATRIC VISCERAL ADIPOSITY INDEX ADAPTATION CORRELATES WITH HOMA-IR, MATSUDA, AND TRANSAMINASES.

    Science.gov (United States)

    Hernández, María José Garcés; Klünder, Miguel; Nieto, Nayely Garibay; Alvarenga, Juan Carlos López; Gil, Jenny Vilchis; Huerta, Samuel Flores; Siccha, Rosa Quispe; Hernandez, Joselin

    2018-03-01

    Visceral adiposity index (VAI) is a mathematical model associated with cardiometabolic risk in adults, but studies on children failed to support this association. Our group has proposed a pediatric VAI model using pediatric ranges, but it has not yet been evaluated and needs further adjustments. The objective of this study was to further adjust the proposed pediatric VAI by age, creating a new pediatric metabolic index (PMI), and assess the correlation of the PMI with insulin resistance indexes and hepatic enzymes. A cross-sectional design with data from 396 children (age 5 to 17 years) was analyzed with a generalized linear model to find the coefficients for triglycerides, high-density-lipoprotein cholesterol, and waist circumference-body mass index quotient. The model was constructed according to sex and age and designated PMI. A cross-validation analysis was performed and a receiver operating characteristic curve was used to determine cut-off points. Significant moderate correlation was found between PMI and homeostatic model assessment of insulin resistance (HOMA-IR) ( r = 0.452; P = .003), Matsuda ( r = -0.366; P = .019), alanine aminotransferase ( r = 0.315, P = .045), and γ-glutamyltransferase ( r = 0.397; P = .010). A PMI score >1.7 was considered as risk. PMI correlates with HOMA-IR, Matsuda, and hepatic enzymes. It could be helpful for identifying children at risk for cardiometabolic diseases. ALT = alanine transaminase BMI = body mass index GGT = γ-glutamyltransferase HDL-C = high-density-lipoprotein cholesterol HOMA-IR = homeostatic model assessment of insulin resistance hs-CRP = high sensitivity C-reactive protein ISI = insulin sensitivity index NAFLD = nonalcoholic fatty liver disease PMI = pediatric metabolic index QUICKI = quantitative insulin sensitivity check index ROC = receiver operating characteristic TG = triglyceride TNF-α = tumor necrosis factor-alpha VAI = visceral adiposity index VAT = visceral adipose tissue WC = waist circumference.

  11. Assessment of Metabolic Changes in Mycobacterium smegmatis Wild-Type and alr Mutant Strains: Evidence of a New Pathway of d-Alanine Biosynthesis.

    Science.gov (United States)

    Marshall, Darrell D; Halouska, Steven; Zinniel, Denise K; Fenton, Robert J; Kenealy, Katie; Chahal, Harpreet K; Rathnaiah, Govardhan; Barletta, Raúl G; Powers, Robert

    2017-03-03

    In mycobacteria, d-alanine is an essential precursor for peptidoglycan biosynthesis. The only confirmed enzymatic pathway to form d-alanine is through the racemization of l-alanine by alanine racemase (Alr, EC 5.1.1.1). Nevertheless, the essentiality of Alr in Mycobacterium tuberculosis and Mycobacterium smegmatis for cell survivability in the absence of d-alanine has been a point of controversy with contradictory results reported in the literature. To address this issue, we examined the effects of alr inactivation on the cellular metabolism of M. smegmatis. The M. smegmatis alr insertion mutant TAM23 exhibited essentially identical growth to wild-type mc 2 155 in the absence of d-alanine. NMR metabolomics revealed drastically distinct phenotypes between mc 2 155 and TAM23. A metabolic switch was observed for TAM23 as a function of supplemented d-alanine. In the absence of d-alanine, the metabolic response directed carbon through an unidentified transaminase to provide the essential d-alanine required for survival. The process is reversed when d-alanine is available, in which the d-alanine is directed to peptidoglycan biosynthesis. Our results provide further support for the hypothesis that Alr is not an essential function of M. smegmatis and that specific Alr inhibitors will have no bactericidal action.

  12. 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)

  13. Carnosine and anserine homeostasis in skeletal muscle and heart is controlled by β‐alanine transamination

    Science.gov (United States)

    Blancquaert, Laura; Baba, Shahid P.; Kwiatkowski, Sebastian; Stautemas, Jan; Stegen, Sanne; Barbaresi, Silvia; Chung, Weiliang; Boakye, Adjoa A.; Hoetker, J. David; Bhatnagar, Aruni; Delanghe, Joris; Vanheel, Bert; Veiga‐da‐Cunha, Maria; Derave, Wim

    2016-01-01

    Key points Using recombinant DNA technology, the present study provides the first strong and direct evidence indicating that β‐alanine is an efficient substrate for the mammalian transaminating enzymes 4‐aminobutyrate‐2‐oxoglutarate transaminase and alanine‐glyoxylate transaminase.The concentration of carnosine and anserine in murine skeletal and heart muscle depends on circulating availability of β‐alanine, which is in turn controlled by degradation of β‐alanine in liver and kidney.Chronic oral β‐alanine supplementation is a popular ergogenic strategy in sports because it can increase the intracellular carnosine concentration and subsequently improve the performance of high‐intensity exercises. The present study can partly explain why the β‐alanine supplementation protocol is so inefficient, by demonstrating that exogenous β‐alanine can be effectively routed toward oxidation. Abstract The metabolic fate of orally ingested β‐alanine is largely unknown. Chronic β‐alanine supplementation is becoming increasingly popular for improving high‐intensity exercise performance because it is the rate‐limiting precursor of the dipeptide carnosine (β‐alanyl‐l‐histidine) in muscle. However, only a small fraction (3–6%) of the ingested β‐alanine is used for carnosine synthesis. Thus, the present study aimed to investigate the putative contribution of two β‐alanine transamination enzymes, namely 4‐aminobutyrate‐2‐oxoglutarate transaminase (GABA‐T) and alanine‐glyoxylate transaminase (AGXT2), to the homeostasis of carnosine and its methylated analogue anserine. We found that, when transfected into HEK293T cells, recombinant mouse and human GABA‐T and AGXT2 are able to transaminate β‐alanine efficiently. The reaction catalysed by GABA‐T is inhibited by vigabatrin, whereas both GABA‐T and AGXT2 activity is inhibited by aminooxyacetic acid (AOA). Both GABA‐T and AGXT2 are highly expressed in the mouse liver and

  14. Crystal Structures of Aedes Aegypt Alanine Glyoxylate Aminotransferase

    Energy Technology Data Exchange (ETDEWEB)

    Han,Q.; Robinson, H.; Gao, Y.; Vogelaar, N.; Wilson, S.; Rizzi, M.; Li, J.

    2006-01-01

    Mosquitoes are unique in having evolved two alanine glyoxylate aminotransferases (AGTs). One is 3-hydroxykynurenine transaminase (HKT), which is primarily responsible for catalyzing the transamination of 3-hydroxykynurenine (3-HK) to xanthurenic acid (XA). Interestingly, XA is used by malaria parasites as a chemical trigger for their development within the mosquito. This 3-HK to XA conversion is considered the major mechanism mosquitoes use to detoxify the chemically reactive and potentially toxic 3-HK. The other AGT is a typical dipteran insect AGT and is specific for converting glyoxylic acid to glycine. Here we report the 1.75{angstrom} high-resolution three-dimensional crystal structure of AGT from the mosquito Aedes aegypti (AeAGT) and structures of its complexes with reactants glyoxylic acid and alanine at 1.75 and 2.1{angstrom} resolution, respectively. This is the first time that the three-dimensional crystal structures of an AGT with its amino acceptor, glyoxylic acid, and amino donor, alanine, have been determined. The protein is dimeric and adopts the type I-fold of pyridoxal 5-phosphate (PLP)-dependent aminotransferases. The PLP co-factor is covalently bound to the active site in the crystal structure, and its binding site is similar to those of other AGTs. The comparison of the AeAGT-glyoxylic acid structure with other AGT structures revealed that these glyoxylic acid binding residues are conserved in most AGTs. Comparison of the AeAGT-alanine structure with that of the Anopheles HKT-inhibitor complex suggests that a Ser-Asn-Phe motif in the latter may be responsible for the substrate specificity of HKT enzymes for 3-HK.

  15. Process considerations for protein engineering of ω-Transaminase

    DEFF Research Database (Denmark)

    Lima Afonso Neto, Watson; Schwarze, Daniel; Tufvesson, Pär

    . However, often wild type enzyme does not fit the requirements of the process conditions, where high substrate and product concentrations as well as high productivity demands of the catalyst (g product per g biocatalyst), are key to economic feasibility. The question thus arises whether to fit the process...... to a wild type transaminase through protein engineering changed the characteristics of the biocatalyst and the implications this would have on a process. A methodology for characterizing the biocatalyst was developed which was subsequently applied to the wild type and 5 mutants selected. It was seen...

  16. Glutamate-pyruvate transaminase subtypes in Singapore ethnic groups.

    Science.gov (United States)

    Saha, N; Bhattacharyya, S P

    1989-01-01

    Autopsy liver samples from 244 Chinese, 119 Malays and 136 Indians were screened for glutamate-pyruvate transaminase (GPT) subtypes by starch-gel electrophoresis and isoelectric focusing at pH 5-7. Altogether, ten phenotypes controlled by four alleles (GPT1, GPT2A, GPT2B and GPT3) were identified. There was no significant difference in the frequency of GPT alleles between the ethnic groups. The distribution of GPT types was in agreement with the Hardy-Weinberg equilibrium in all the ethnic groups.

  17. Applying Enzymatic Cascades for ISCPR in ω-transaminase Systems

    DEFF Research Database (Denmark)

    Janes, Kresimir; Woodley, John; Tufvesson, Pär

    steps. An important fact is also that different types of selectivity make biocatalysis an excellent tool for overcoming difficulties typically associated with organic synthesis. Regioselectivity of the biocatalysts offers potential process simplification compared to the organic synthesis routes......Filtration Membrane Reactor) as a viable process design option and charge analysis showed that ISPR is possible via ion exchange resins or electrodialysis. An ISPR example showed that process intensification could yield significant reductions in the required ω-transaminase activity improvement (up to five fold...

  18. Insulin-induced inhibition of gluconeogenesis genes, including glutamic pyruvic transaminase 2, is associated with reduced histone acetylation in a human liver cell line.

    Science.gov (United States)

    Honma, Kazue; Kamikubo, Michiko; Mochizuki, Kazuki; Goda, Toshinao

    2017-06-01

    Hepatic glutamic pyruvic transaminase (GPT; also known as alanine aminotransferase) is a gluconeogenesis enzyme that catalyzes conversions between alanine and pyruvic acid. It is also used as a blood biomarker for hepatic damage. In this study, we investigated whether insulin regulates GPT expression, as it does for other gluconeogenesis genes, and if this involves the epigenetic modification of histone acetylation. Human liver-derived HepG2 cells were cultured with 0.5-100nM insulin for 8h, and the mRNA expression of GPT, glutamic-oxaloacetic transaminase (GOT), γ-glutamyltransferase (GGT), PCK1, G6PC and FBP1 was measured. We also investigated the extent of histone acetylation around these genes. Insulin suppressed the mRNA expression of gluconeogenesis genes (GPT2, GOT1, GOT2, GGT1, GGT2, G6PC, and PCK1) in HepG2 cells in a dose-dependent manner. mRNA levels of GPT2, but not GPT1, were decreased by insulin. Histone acetylation was also reduced around GPT2, G6PC, and PCK1 in response to insulin. The expression of GPT2 and other gluconeogenesis genes such as G6PC and PCK1 was suppressed by insulin, in association with decreases in histone H3 and H4 acetylation surrounding these genes. Copyright © 2017 Elsevier Inc. All rights reserved.

  19. 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)

  20. 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

  1. 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...

  2. ?-Alanine supplementation and military performance

    OpenAIRE

    Hoffman, Jay R.; Stout, Jeffrey R.; Harris, Roger C.; Moran, Daniel S.

    2015-01-01

    During sustained high-intensity military training or simulated combat exercises, significant decreases in physical performance measures are often seen. The use of dietary supplements is becoming increasingly popular among military personnel, with more than half of the US soldiers deployed or garrisoned reported to using dietary supplements. ?-Alanine is a popular supplement used primarily by strength and power athletes to enhance performance, as well as training aimed at improving muscle grow...

  3. Changes of liver fat content and transaminases in obese children after 12-mo nutritional intervention.

    Science.gov (United States)

    Verduci, Elvira; Pozzato, Carlo; Banderali, Giuseppe; Radaelli, Giovanni; Arrizza, Chiara; Rovere, Antonio; Riva, Enrica; Giovannini, Marcello

    2013-09-27

    To assess a relationship between longitudinal changes in liver fat content and biochemical parameters in obese children after 1-year nutritional intervention. Forty-six obese children, 21 males and 25 females, aged 6-14 years, underwent metabolic measurements, liver ultrasonography (US) and chemical-shift magnetic resonance imaging (MRI) examinations at baseline and after 1-year nutritional intervention. A child was defined obese if her/his body mass index (BMI) was above the age- and sex-adjusted BMI Cole's curve passing through the cut-off of 30 kg/m(2) at 18 years. BMI Z scores were calculated and adjusted for age and gender by using the Cole's LMS-method and Italian reference data. Biochemistry included serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST). Abdominal US and chemical-shift MRI were performed according to a randomized sequence. The same radiologist performed US by a GE Logiq 9 (General Electric Healthcare Medical Systems, Milwaukee, WI, United States) using a 3.5-MHz convex array transducer. Liver echogenicity was evaluated independently on videotape by 3 radiologists unaware of the child and MRI outcomes, and a consensus was established. Another experienced radiologist, unaware of the child and US data, performed the abdominal chemical-shift MRI with a 1-t system NT-Intera (Philips Medical Systems, Best, The Netherlands) and a phased-array coil. Liver fat fraction (FF) on MRI was judged elevated when greater than 9%. A FF > 18% was considered expressing more severe cases of fatty liver according to Fishbein. A nutritional-behavioral intervention was recommended to promote a normocaloric balanced diet and active lifestyle based on the Italian guidelines for treatment of childhood obesity. Compared to baseline, at the end of intervention children showed lower intakes of energy (mean ± SD: 2549 ± 1238 Kcal vs 1770 ± 622 Kcal, P intervention association was found with AST (P = 0.017). Change of FF was associated with change in

  4. Vibrational dynamics of crystalline L-alanine

    Energy Technology Data Exchange (ETDEWEB)

    Bordallo, H.N.; Eckert, J. [Los Alamos National Lab., NM (United States); Barthes, M. [Univ. Montpellier II (France)

    1997-11-01

    The authors report a new, complete vibrational analysis of L-alanine and L-alanine-d{sub 4} which utilizes IINS intensities in addition to frequency information. The use of both isotopomers resulted in a self-consistent force field for and assignment of the molecular vibrations in L-alanine. Some details of the calculation as well as a comparison of calculated and observed IINS spectra are presented. The study clarifies a number of important issues on the vibrational dynamics of this molecule and presents a self-consistent force field for the molecular vibrations in crystalline L-alanine.

  5. Contribution of the GABA shunt to hypoxia-induced alanine accumulation in roots of Arabidopsis thaliana.

    Science.gov (United States)

    Miyashita, Yo; Good, Allen G

    2008-01-01

    When subjected to low oxygen stress, plants accumulate alanine and gamma-aminobutyric acid (GABA). To investigate the function of GABA metabolism under hypoxia and its contribution to alanine accumulation, we studied the genes that encode the two key enzymes of the GABA shunt, glutamate decarboxylase (GAD) and GABA transaminase (GABA-T). Among the five homologous GAD genes found in Arabidopsis thaliana, GAD1 expression was predominantly found in roots, while GAD2 expression was evident in all organs. Expression of the other three GAD genes was generally weak. In response to hypoxia, transcriptional induction was observed for GAD4 only. For GABA-T1, its expression was detected in all organs, but there was no significant transcriptional change under hypoxic conditions. Moreover, we have isolated and characterized Arabidopsis mutants defective in GAD1 and GABA-T1. In gad1 mutants, GAD activity was significantly reduced in roots but was not affected in shoots. In the gaba-t1 mutant, GABA-T activity was decreased to negligible levels in both shoots and roots. These mutants were phenotypically normal under normal growth conditions except for the reduced seed production of the pop2 mutants as described previously. However, metabolite analysis revealed significant changes in GABA content in gad1 and gaba-t1 mutants. The levels of alanine under hypoxic conditions were also affected in the roots of gad1 and gaba-t1 mutants. The partial inhibition of the hypoxia-induced alanine accumulation in roots of these mutants suggests that the GABA shunt is, in part, responsible for the alanine accumulation under hypoxia.

  6. Catalytic Stereoinversion of L-Alanine to Deuterated D-Alanine.

    Science.gov (United States)

    Moozeh, Kimia; So, Soon Mog; Chin, Jik

    2015-08-03

    A combination of an achiral pyridoxal analogue and a chiral base has been developed for catalytic deuteration of L-alanine with inversion of stereochemistry to give deuterated D-alanine under mild conditions (neutral pD and 25 °C) without the use of any protecting groups. This system can also be used for catalytic deuteration of D-alanine with retention of stereochemistry to give deuterated D-alanine. Thus a racemic mixture of alanine can be catalytically deuterated to give an enantiomeric excess of deuterated D-alanine. While catalytic deracemization of alanine is forbidden by the second law of thermodynamics, this system can be used for catalytic deracemization of alanine with deuteration. Such green and biomimetic approach to catalytic stereocontrol provides insights into efficient amino acid transformations. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  7. 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

  8. 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

  9. 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 ...

  10. Hepatic gluconeogenesis from alanine following surgery.

    Science.gov (United States)

    Yoshikawa, K; Setsu, M; Mishina, T; Koyama, S; Muto, T

    1982-01-01

    To clarify alanine metabolism in the liver with special reference to gluconeogenesis, catheters were placed in hepatic, portal, femoral, and external jugular veins of six male mongrel dogs. A trace amount of 14C-alanine was administered as a single pulse into the external jugular vein on the first postoperative day, and the blood samples were taken from each vein for the subsequent two hours to measure 14C-glucose radioactivity. Cumulative radioactivity after 14C-alanine injection showed that 74 per cent of the radioactivity in whole protein-free serum was that of 14C-glucose. Therefore, it is considered that the metabolic pathway of alanine in the liver after surgery is mainly through gluconeogenesis to glucose. Our in vivo experiment clearly showed that hepatic gluconeogenesis from alanine is one of the important factors related to hyperglycemia after surgery.

  11. 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.

  12. 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)

  13. 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.

  14. Implementation of an alanine dosimetry service

    International Nuclear Information System (INIS)

    Gago Arias, A.; Nunez Pelaez, N.; Peteiro Vilaseco, E.; Gomez Rodriguez, F.; Gonzalez Castano, D. M.

    2011-01-01

    This work facing the implementation of an alanine dosimetry service, linked to the installation of Co 6 0 Radio physics Laboratory (LP) and Paramagnetic Resonance Service of the University of Santiago de Compostela (USC).

  15. 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

  16. Solved? The reductive radiation chemistry of alanine.

    Science.gov (United States)

    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.

  17. β-Alanine supplementation and military performance.

    Science.gov (United States)

    Hoffman, Jay R; Stout, Jeffrey R; Harris, Roger C; Moran, Daniel S

    2015-12-01

    During sustained high-intensity military training or simulated combat exercises, significant decreases in physical performance measures are often seen. The use of dietary supplements is becoming increasingly popular among military personnel, with more than half of the US soldiers deployed or garrisoned reported to using dietary supplements. β-Alanine is a popular supplement used primarily by strength and power athletes to enhance performance, as well as training aimed at improving muscle growth, strength and power. However, there is limited research examining the efficacy of β-alanine in soldiers conducting operationally relevant tasks. The gains brought about by β-alanine use by selected competitive athletes appears to be relevant also for certain physiological demands common to military personnel during part of their training program. Medical and health personnel within the military are expected to extrapolate and implement relevant knowledge and doctrine from research performed on other population groups. The evidence supporting the use of β-alanine in competitive and recreational athletic populations suggests that similar benefits would also be observed among tactical athletes. However, recent studies in military personnel have provided direct evidence supporting the use of β-alanine supplementation for enhancing combat-specific performance. This appears to be most relevant for high-intensity activities lasting 60-300 s. Further, limited evidence has recently been presented suggesting that β-alanine supplementation may enhance cognitive function and promote resiliency during highly stressful situations.

  18. 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...

  19. 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

  20. A Rapid Selection Procedure for Simple Commercial Implementation of omega-Transaminase Reactions

    DEFF Research Database (Denmark)

    Gundersen Deslauriers, Maria; Tufvesson, Pär; Rackham, Emma J.

    2016-01-01

    A stepwise selection procedure is presented to quickly evaluate whether a given omega-transaminase reaction is suitable for a so-called "simple" scale-up for fast industrial implementation. Here "simple" is defined as a system without the need for extensive process development or specialized...

  1. Nitrogen metabolism in actinorhizal nodules of Alnus glutinosa: expression of glutamine synthetase and acetylornithine transaminase.

    NARCIS (Netherlands)

    Guan, C.; Ribeiro, A.; Akkermans, A.D.L.; Jing, Y.; Kammen, van A.; Bisseling, T.; Pawlowski, K.

    1996-01-01

    Two nodule cDNA clones representing genes involved in Alnus glutinosa nitrogen metabolism were analysed. ag11 encoded glutamine synthetase (GS), the enzyme responsible for ammonium assimilation, while ag118 encoded acetylornithine transaminase (AOTA), an enzyme involved in the biosynthesis of

  2. A Practical and Fast Method To Predict the Thermodynamic Preference of omega-Transaminase-Based Transformations

    DEFF Research Database (Denmark)

    Meier, Robert J.; Gundersen Deslauriers, Maria; Woodley, John

    2015-01-01

    A simple, easy-to-use, and fast approach method is proposed and validated that can predict whether a transaminase reaction is thermodynamically unfavourable. This allowed us to de-select, in the present case, at least 50% of the reactions because they were thermodynamically unfavourable as confir...

  3. 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...

  4. The effect of alanine on glucagon secretion

    Science.gov (United States)

    Müller, Walter A.; Faloona, Gerald R.; Unger, Roger H.

    1971-01-01

    If glucagon plays a hormonal role in the regulation of gluconeogenesis from endogenous amino acids, its secretion might be stimulated by an increase in the concentration of alanine, which has recently been identified as a principal gluconeogenic precursor. To determine if this is the case, 0.75 mmole of alanine per kilo was infused into conscious dogs immediately after a priming injection of 0.25 mmole per kg for 15 min. A uniform rise in the plasma level of pancreatic glucagon, as determined by a relatively specific radioimmunoassay for pancreatic glucagon, was observed. The rise, which averaged 90 pg per ml, was highly significant at 7½ and 15 min after the start of the infusion. Insulin rose an average of only 8 μU per ml, while glucose rose an average of 10 mg per 100 ml. A lower dose of alanine, 1 mmole per kg, infused over a 1 hr period without an initial priming injection, also elicited a significant rise in glucagon measured in the pancreaticoduodenal venous plasma; glucagon rose from 350 pg per ml to 1066 pg per ml at the end of the infusion. The insulin response was modest and inconsistent, and glucose, again, rose 10 mg per 100 ml. To determine if the availability of exogenous glucose would abolish the alanine-induced rise in glucagon secretion, dogs were made hyperglycemic by a constant intravenous glucose infusion and were then given the high-dose alanine infusion. Under these circumstances, glucagon did not rise above the mean fasting concentration of 75 pg per ml, whereas mean insulin rose dramatically by more than 100 μU per ml. It was concluded that, in the fasting state, alanine does stimulate the secretion of glucagon, while having very little stimulatory effect on insulin secretion. Glucagon could, therefore, be a humoral mediator of gluconeogenesis from endogenous alanine, responding to hyperalaninemia in the fasting state, but not when exogenous glucose is available. PMID:5116210

  5. Identification of branched-chain amino acid aminotransferases active towards (R)-(+)-1-phenylethylamine among PLP fold type IV transaminases.

    Science.gov (United States)

    Bezsudnova, Ekaterina Yu; Dibrova, Daria V; Nikolaeva, Alena Yu; Rakitina, Tatiana V; Popov, Vladimir O

    2018-04-10

    New class IV transaminases with activity towards L-Leu, which is typical of branched-chain amino acid aminotransferases (BCAT), and with activity towards (R)-(+)-1-phenylethylamine ((R)-PEA), which is typical of (R)-selective (R)-amine:pyruvate transaminases, were identified by bioinformatics analysis, obtained in recombinant form, and analyzed. The values of catalytic activities in the reaction with L-Leu and (R)-PEA are comparable to those measured for characteristic transaminases with the corresponding specificity. Earlier, (R)-selective class IV transaminases were found to be active, apart from (R)-PEA, only with some other (R)-primary amines and D-amino acids. Sequences encoding new transaminases with mixed type of activity were found by searching for changes in the conserved motifs of sequences of BCAT by different bioinformatics tools. Copyright © 2018 Elsevier B.V. All rights reserved.

  6. 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...

  7. A molecular receptor selective for zwitterionic alanine.

    Science.gov (United States)

    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.

  8. Alanine racemase from the acidophile Acetobacter aceti.

    Science.gov (United States)

    Francois, Julie A; Kappock, T Joseph

    2007-01-01

    Acetobacter aceti converts ethanol to acetic acid, and survives acetic acid exposure by tolerating cytoplasmic acidification. Alanine racemase (Alr) is a pyridoxal 5' phosphate (PLP) -dependent enzyme that catalyzes the interconversion of the d- and l-isomers of alanine and has a basic pH optimum. Since d-alanine is essential for peptidoglycan biosynthesis, Alr must somehow function in the acidic cytoplasm of A. aceti. We report the partial purification of native A. aceti Alr (AaAlr) and evidence that it is a rather stable enzyme. The C-terminus of AaAlr has a strong resemblance to the ssrA-encoded protein degradation signal, which thwarted initial protein expression experiments. High-activity AaAlr forms lacking a protease recognition sequence were expressed in Escherichia coli and purified. Biophysical and enzymological experiments confirm that AaAlr is intrinsically acid-resistant, yet has the catalytic properties of an ordinary Alr.

  9. Changes of ammonia, urea contents and transaminase activity in the body during aerial exposure and ammonia loading in Chinese loach Paramisgurnus dabryanus.

    Science.gov (United States)

    Zhang, Yun-Long; Zhang, Hai-Long; Wang, Ling-Yu; Gu, Bei-Yi; Fan, Qi-Xue

    2017-04-01

    The Paramisgurnus dabryanus was exposed to 30 mmol L -1 NH 4 Cl solution and air to assessing the change of body ammonia and urea contents and the activities of alanine aminotransferase (ALT) and aspartate transaminase (AST). After 48 h of ammonia exposure, ammonia concentration in the plasma, brain, liver and muscle were 3.3-fold, 5.6-fold, 3.5-fold and 4.2-fold, respectively, those of the control values. Plasma, brain, liver and muscle ammonia concentrations increased to 2.2-fold, 3.3-fold, 2.5-fold and 2.9-fold, respectively, those of control values in response to 48 h of aerial exposure. Within the given treatment (ammonia or aerial exposure), there was no change in plasma, brain and liver urea concentrations between exposure durations. The plasma ALT activity was significantly affected by exposure time during aerial exposure, while the liver ALT activity was not affected by ammonia or aerial exposure. Exposure to NH 4 Cl or air had no effect on either plasma or liver AST activity. Our results suggested that P. dabryanus could accumulate quite high level of internal ammonia because of the high ammonia tolerance in its cells and tissues, and NH 3 volatilization would be a possible ammonia detoxification strategy in P. dabryanus. Urea synthesis was not an effective mechanism to deal with environmental or internal ammonia problem. The significant increase of ALT activity in plasma during aerial exposure, indicating that alanine synthesis through certain amino acid catabolism may be subsistent in P. dabryanus.

  10. 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)

  11. Dose Response of Alanine Detectors Irradiated with Carbon Ion Beams

    DEFF Research Database (Denmark)

    Herrmann, Rochus; Jäkel, Oliver; Palmans, Hugo

    2011-01-01

    Purpose: The dose response of the alanine detector shows a dependence on particle energy and type, when irradiated with ion beams. The purpose of this study is to investigate the response behaviour of the alanine detector in clinical carbon ion beams and compare the results with model predictions....... Methods: Alanine detectors have been irradiated with carbon ions with an energy range of 89-400 MeV/u. The relative effectiveness of alanine has been measured in this regime. Pristine and spread out Bragg peak depth-dose curves have been measured with alanine dosimeters. The track-structure based alanine...

  12. Potential energy surface of alanine polypeptide chains

    DEFF Research Database (Denmark)

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

    2006-01-01

    The multidimensional potential energy surfaces of the peptide chains consisting of three and six alanine (Ala) residues have been studied with respect to the degrees of freedom related to the twist of these molecules relative to the peptide backbone (these degrees of freedom are responsible...

  13. Ursodeoxycholic Acid Can Improve Liver Transaminase Quantities in Children with Anticonvulsant Drugs Hepatotoxicity: a Pilot Study.

    Directory of Open Access Journals (Sweden)

    Masoumeh Asgarshirazi

    2015-06-01

    Full Text Available The present study has been directed to investigate Ursodeoxycholic Acid (UDCA effect in children, to reduce the high Liver transaminases induced by Anticonvulsant drugs (drug induced hepatitis. This idea has been driven from Cytoprotective and antioxidant properties of UDCA to be used in drug induced inflammation in Liver. Twenty two epileptic patients aged between 4 mo - 3 yr whom were under anticonvulsant therapy with drugs such as valperoic acid, primidone, levetiracetam, Phenobarbital or any combination of them and had shown Liver transaminases rise , after rule out of Viral-Autoimmune, Metabolic and Anatomic causes, have been prescribed UDCA in dose of 10-15 mg/kg/day, at least for 6 months. Any patient who have shown confusing factors such as genetic disorders with liver involvement or spontaneous decline in enzymes or had not treatment compliance has been excluded from the study. Transaminases range changes as well as Probable side effects of the drug have been monitored. The results indicated that UDCA is effective and well tolerable in the children with drug induced hyper transaminasemia. No side effect has been seen and recorded in this study. Based on this study and its results, we recommend UDCA as a safe and effective choice in drug induced hepatotoxicities.

  14. Response of regional brain glutamate transaminases of rat to aluminum in protein malnutrition

    Directory of Open Access Journals (Sweden)

    Chatterjee Ajay K

    2002-08-01

    Full Text Available Abstract Background The mechanism of aluminum-induced neurotoxicity is not clear. The involvement of glutamate in the aluminium-induced neurocomplications has been suggested. Brain glutamate levels also found to be altered in protein malnutrition. Alterations in glutamate levels as well as glutamate-α-decarboxylase in different regions of rat brain has been reported in response to aluminum exposure. Thus the study of glutamate metabolising enzymes in different brain regions of rats maintained on either normal or restricted protein diet may be of importance for understanding the neurotoxicity properties of aluminium. Results Dietary protein restrictions does not have an significant impact on regional aluminum content of the brain. The interaction of aluminum intoxication and protein restriction is significant in the thalamic area and the midbrain-hippocampal region in cases of glutamate oxaloacetate transaminase. In the case of gluatmate pyruvate transaminase, this interaction is significant only in thalamic area. Conclusion The metabolism of amino acids, as indicated by activities of specific transaminases, of brain is altered in response to aluminum exposure. These alterations are region specific and are dependent on dietary protein intake or manipulation of the brain amino acid homeostasis.

  15. Response of regional brain glutamate transaminases of rat to aluminum in protein malnutrition

    Science.gov (United States)

    Nayak, Prasunpriya; Chatterjee, Ajay K

    2002-01-01

    Background The mechanism of aluminum-induced neurotoxicity is not clear. The involvement of glutamate in the aluminium-induced neurocomplications has been suggested. Brain glutamate levels also found to be altered in protein malnutrition. Alterations in glutamate levels as well as glutamate-α-decarboxylase in different regions of rat brain has been reported in response to aluminum exposure. Thus the study of glutamate metabolising enzymes in different brain regions of rats maintained on either normal or restricted protein diet may be of importance for understanding the neurotoxicity properties of aluminium. Results Dietary protein restrictions does not have an significant impact on regional aluminum content of the brain. The interaction of aluminum intoxication and protein restriction is significant in the thalamic area and the midbrain-hippocampal region in cases of glutamate oxaloacetate transaminase. In the case of gluatmate pyruvate transaminase, this interaction is significant only in thalamic area. Conclusion The metabolism of amino acids, as indicated by activities of specific transaminases, of brain is altered in response to aluminum exposure. These alterations are region specific and are dependent on dietary protein intake or manipulation of the brain amino acid homeostasis. PMID:12197946

  16. 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

  17. Increase in transaminase levels following the use of raltegravir in a woman with a high HIV viral load at 35 weeks of pregnancy.

    Science.gov (United States)

    Renet, Sophie; Closon, Aurélie; Brochet, Marie-Sophie; Bussières, Jean-François; Boucher, Marc

    2013-01-01

    Despite the efficacy of raltegravir in reducing viral load in HIV-infected patients, evidence for its safety in late pregnancy is lacking. A high rate of placental transfer was recently demonstrated. A treatment-naïve 34-year-old HIV-1-positive woman of African origin began treatment with zidovudine/lamivudine, lopinavir/ritonavir, and raltegravir at 35 weeks of pregnancy. After 11 days of treatment with raltegravir, a substantial reduction in viral load was achieved. Concurrently, she had a 23-fold increase in serum alanine aminotransferase and a 10-fold increase in serum aspartate aminotransferase, both of which returned to normal when raltegravir treatment was discontinued. A healthy boy was delivered at term. The infant's tests for HIV were negative at five months, and he had no health problems at eight months. This is the first case report, to our knowledge, of increased maternal serum transaminase levels following the use of raltegravir in a woman at a late stage of pregnancy.

  18. Secretion of d-alanine by Escherichia coli.

    Science.gov (United States)

    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.

  19. 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.

  20. 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.)

  1. 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.)

  2. Amine donor and acceptor influence on the thermodynamics of ω-transaminase reactions

    DEFF Research Database (Denmark)

    Gundersen, Maria T.; Abu, Rohana; Schürmann, Martin

    2015-01-01

    In recent years biocatalytic transamination using ω-transaminase has become established as one of the most interesting routes to synthesize chiral amines with a high enantiomeric purity, especially in the pharmaceutical sector where the demand for such compounds is high. Nevertheless, one....... It was found that an aliphatic six membered ring was favourable and a conjugated bicyclic five membered ring structure, unfavourable. Finally, we evaluated and compared the use of five different donor molecules, and calculated their ΔGapp values. This is particularly important in the further implementation...

  3. Immobilization of Escherichia coli containing ω‐transaminase activity in LentiKats®

    DEFF Research Database (Denmark)

    Cárdenas‐Fernández, Max; Lima Afonso Neto, Watson; López, Carmen

    2012-01-01

    Whole Escherichia coli cells overexpressing ω‐transaminase (ω‐TA) and immobilized cells entrapped in LentiKats® were used as biocatalysts in the asymmetric synthesis of the aromatic chiral amines 1‐phenylethylamine (PEA) and 3‐amino‐1‐phenylbutane (APB). Whole cells were permeabilized...... whole cell biocatalysis, the reaction with IPA was one order of magnitude faster than with Ala. No reaction was detected when permeabilized E. coli cells containing ω‐TA were employed using Ala as the amino donor. Additionally, the synthesis of APB from 4‐phenyl‐2‐butanone and IPA was studied. Whole...

  4. Nucleic acids encoding plant glutamine phenylpyruvate transaminase (GPT) and uses thereof

    Science.gov (United States)

    Unkefer, Pat J.; Anderson, Penelope S.; Knight, Thomas J.

    2016-03-29

    Glutamine phenylpyruvate transaminase (GPT) proteins, nucleic acid molecules encoding GPT proteins, and uses thereof are disclosed. Provided herein are various GPT proteins and GPT gene coding sequences isolated from a number of plant species. As disclosed herein, GPT proteins share remarkable structural similarity within plant species, and are active in catalyzing the synthesis of 2-hydroxy-5-oxoproline (2-oxoglutaramate), a powerful signal metabolite which regulates the function of a large number of genes involved in the photosynthesis apparatus, carbon fixation and nitrogen metabolism.

  5. Towards a standardized way of reporting physicochemical data and process metrics for transaminase reactions

    DEFF Research Database (Denmark)

    Tufvesson, Pär; Woodley, John

    information about the system and the reaction performance is lacking in the otherwise very useful scientific reports at laboratory scale. For instance, although K equilibrium is one of the key determining factors for the design and scale-up any transaminase process, it is very rarely reported [3]. In order...... to build a broader understanding of the correlation between the underlying physicochemical properties of the system (e.g. substrate volatility) and the process performance (e.g. gram of product per gram of biocatalyst), it would be highly beneficial if these data were reported, and ideally in a consistent...

  6. 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.

  7. 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)

  8. 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)

  9. 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

  10. Glutamate oxaloacetate transaminase enables anaplerotic refilling of TCA cycle intermediates in stroke-affected brain.

    Science.gov (United States)

    Rink, Cameron; Gnyawali, Surya; Stewart, Richard; Teplitsky, Seth; Harris, Hallie; Roy, Sashwati; Sen, Chandan K; Khanna, Savita

    2017-04-01

    Ischemic stroke results in excessive release of glutamate, which contributes to neuronal cell death. Here, we test the hypothesis that otherwise neurotoxic glutamate can be productively metabolized by glutamate oxaloacetate transaminase (GOT) to maintain cellular energetics and protect the brain from ischemic stroke injury. The GOT-dependent metabolism of glutamate was studied in primary neural cells and in stroke-affected C57-BL6 mice using magnetic resonance spectroscopy and GC-MS. Extracellular Glu sustained cell viability under hypoglycemic conditions and increased GOT-mediated metabolism in vitro Correction of stroke-induced hypoxia using supplemental oxygen in vivo lowered Glu levels as measured by 1 H magnetic resonance spectroscopy. GOT knockdown abrogated this effect and caused ATP loss in the stroke-affected brain. GOT overexpression increased anaplerotic refilling of tricarboxylic acid cycle intermediates in mouse brain during ischemic stroke. Furthermore, GOT overexpression not only reduced ischemic stroke lesion volume but also attenuated neurodegeneration and improved poststroke sensorimotor function. Taken together, our results support a new paradigm that GOT enables metabolism of otherwise neurotoxic extracellular Glu through a truncated tricarboxylic acid cycle under hypoglycemic conditions.-Rink, C., Gnyawali, S., Stewart, R., Teplitsky, S., Harris, H., Roy, S., Sen, C. K., Khanna, S. Glutamate oxaloacetate transaminase enables anaplerotic refilling of TCA cycle intermediates in stroke-affected brain. © FASEB.

  11. Termoativação da transaminase glutâmico oxaloacética

    Directory of Open Access Journals (Sweden)

    Hélion Póvoa Júnior

    1973-01-01

    Full Text Available Estudou-se a atividade da transaminase glutâmico oxaloacética (TGO em diferentes tecidos (fígado, músculo, rim, pulmão, baço e soro sanguíneo de ratos e de soro humano. verificou-se que a atividade da enzima proveniente de qualquer um destes tecidos é aumentada cerca de tr~es vezes quando a incubação se faz a 60ºC, ao invés de 37ºC. São feitas considerações acerca da importância deste fato.Glutamic Oxalacetic transaminase is thermoativated. Its optimum of catalytical activity is at 60ºC. At this temperature, colour is approximately three times more intense than at 37ºC, temperature usually utilized for determination of enzyme activity. This phenomenon is observed in human blood serum and several rat tissues (liver, heart, striated muscle, spleen, lung, kidney and blood serum.

  12. Correlation between hematologic profile and transaminase enzymes with hospitalization duration dengue

    Science.gov (United States)

    Tinambunan, E.; Suryani; Katu, S.; Halim, R.; Mubin, A. H.; Sahyuddin

    2018-03-01

    Dengue is an infectious disease that can be found from mild to severe andaffected the clinical spectrum of the disease. Various hematologic profiles and transaminase enzymes are thought to reflect the severity of the disease thus affecting the hospitalization duration. For determining the correlation between hematological profile and transaminase enzyme to the hospitalization duration in dengue patients, an observational design study with the cross-sectional approach on dengue subjects was from 2 hospitals in Makassar. Hemoglobin, leukocyte, thrombocyte, AST, ALT, PT, and APTT were examined for hospitalization duration. There were 65 samples (34 men, 31 women) with the length of stay dengue patients. There was no correlation between the elevated of hematocrit value (p = 0.429), thrombocytopenia (p = 1.000), elevated of AST (p = 0.456) and ALT (p = 0.285) on hospitalization duration. In conclusion, low leukocyte values and APTT prolongation correlate with hospitalization duration but did not correlate significantly with hospitalization duration for elevated hematocrit, thrombocytopenia, elevated AST, and ALT.

  13. Enantioselective Transamination in Continuous Flow Mode with Transaminase Immobilized in a Macrocellular Silica Monolith

    Directory of Open Access Journals (Sweden)

    Ludivine van den Biggelaar

    2017-02-01

    Full Text Available ω-Transaminases have been immobilized on macrocellular silica monoliths and used as heterogeneous biocatalysts in a continuous flow mode enantioselective transamination reaction. The support was prepared by a sol-gel method based on emulsion templating. The enzyme was immobilized on the structured silica monoliths both by adsorption, and by covalent grafting using amino-functionalized silica monoliths and glutaraldehyde as a coupling agent. A simple reactor set-up based on the use of a heat-shrinkable Teflon tube is presented and successfully used for the continuous flow kinetic resolution of a chiral amine, 4-bromo-α-methylbenzylamine. The porous structure of the supports ensures effective mass transfer and the reactor works in the plug flow regime without preferential flow paths. When immobilized in the monolith and used in the flow reactor, transaminases retain their activity and their enantioselectivity. The solid biocatalyst is also shown to be stable both on stream and during storage. These essential features pave the way to the successful development of an environmentally friendly process for chiral amines production.

  14. Alanine aminotransferase catalyses the breakdown of alanine after hypoxia in Arabidopsis thaliana.

    Science.gov (United States)

    Miyashita, Yo; Dolferus, Rudy; Ismond, Kathleen P; Good, Allen G

    2007-03-01

    Alanine aminotransferase (AlaAT) catalyses the reversible transfer of an amino group from glutamate to pyruvate to form 2-oxoglutarate and alanine. The regulation of AlaAT in several plant species has been studied in response to low-oxygen stress, light and nitrogen application. In this study, induction of Arabidopsis AlaAT1 and AlaAT2 during hypoxia was observed at the transcriptional level, and an increase in enzyme activity was detected in hypoxically treated roots. In addition, the tissue-specific expression of AlaAT1 and AlaAT2 was analysed using promoter:GUS fusions. The GUS staining patterns indicated that both AlaAT genes are expressed predominantly in vascular tissues. We manipulated AlaAT expression to determine the relative importance of this enzyme in low-oxygen stress tolerance and nitrogen metabolism. This was done by analysing T-DNA mutants and over-expressing barley AlaAT in Arabidopsis. The AlaAT1 knockout mutant (alaat1-1) showed a dramatic reduction in AlaAT activity, suggesting that AlaAT1 is the major AlaAT isozyme in Arabidopsis. Over-expression of barley AlaAT significantly increased the AlaAT activity in the transgenic plants. These plants were analysed for metabolic changes over a period of hypoxic stress and during their subsequent recovery. The results showed that alaat1-1 plants accumulate more alanine than wild-type plants during the early phase of hypoxia, and the decline in accumulated alanine was delayed in the alaat1-1 line during the post-hypoxia recovery period. When alanine was supplied as the nitrogen source, alaat1-1 plants utilized alanine less efficiently than wild-type plants did. These results indicate that the primary role of AlaAT1 is to break down alanine when it is in excess. Therefore, AlaAT appears to be crucial for the rapid conversion of alanine to pyruvate during recovery from low-oxygen stress.

  15. 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.

  16. 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

  17. Determination of the dose of traffic in HDR brachytherapy with ALANINE/R PE technique

    International Nuclear Information System (INIS)

    Guzman Calcina, C. S.; Chen, F.; Almeida, A. de; Baffa, O.

    2001-01-01

    It determines, experimentally, the dose of traffic in brachytherapy for High Dose Rate (HDR), using for the first-time the Electronic Paramagnetic Resonance (EPR) technique with alanine detectors. The value obtained is the published next to obtained using lithium fluoride thermoluminescent dosimeters [es

  18. 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

  19. Green reduction of graphene oxide using alanine.

    Science.gov (United States)

    Wang, Jiabin; Salihi, Elif Caliskan; Šiller, Lidija

    2017-03-01

    There remains a real need for the easy, eco-friendly and scalable preparation method of graphene due to various potential applications. Chemical reduction is the most versatile method for the large scale production of graphene. Here we report the operating conditions for a one-step, economical and green synthesis method for the reduction of graphene oxide using a biomolecule (alanine). Graphene oxide was produced by the oxidation and exfoliation of natural graphite flake with strong oxidants using Hummers method (Hummers and Offeman, 1958), but the method was revised in our laboratory to set up a safe and environmentally friendly route. The reduction of graphene oxide was investigated using alanine at various operating conditions in order to set up optimum conditions (treatment time, temperature and concentration of the reagent). Samples have been characterized by using UV-Visible spectroscopy, Fourier transform infrared spectroscopy, transmission electron microscopy, Raman spectroscopy and X-ray diffraction analysis. Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.

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

    Science.gov (United States)

    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.

  1. Biotransformation of β-keto nitriles to chiral (S)-β-amino acids using nitrilase and ω-transaminase.

    Science.gov (United States)

    Mathew, Sam; Nadarajan, Saravanan Prabhu; Sundaramoorthy, Uthayasuriya; Jeon, Hyunwoo; Chung, Taeowan; Yun, Hyungdon

    2017-04-01

    To enzymatically synthesize enantiomerically pure β-amino acids from β-keto nitriles using nitrilase and ω-transaminase. An enzyme cascade system was designed where in β-keto nitriles are initially hydrolyzed to β-keto acids using nitrilase from Bradyrhizobium japonicum and subsequently β-keto acids were converted to β-amino acids using ω-transaminases. Five different ω-transaminases were tested for this cascade reaction, To enhance the yields of β-amino acids, the concentrations of nitrilase and amino donor were optimized. Using this enzymatic reaction, enantiomerically pure (S)-β-amino acids (ee > 99%) were generated. As nitrilase is the bottleneck in this reaction, molecular docking analysis was carried out to depict the poor affinity of nitrilase towards β-keto acids. A novel enzymatic route to generate enantiomerically pure aromatic (S)-β-amino acids from β-keto nitriles is demonstrated for the first time.

  2. High-performance liquid chromatographic analysis of analogous amino and oxo acids for the determination of amino acid oxidase and transaminase activities.

    Science.gov (United States)

    Brodelius, P

    1984-01-01

    An analytical method, based on reversed-phase high-performance liquid chromatography, for the determination of analogous amino and oxo acids, has been developed. The assay may be employed to monitor amino acid oxidase and transaminase activities. Simultaneous quantitative determination of substrate and product is readily achieved. In particular, the amino acid oxidase activity within whole microbial cells immobilized by entrapment in Ca2+-alginate has been studied. The assay of transaminases has been exemplified with glutamic-pyruvic transaminase.

  3. 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.

  4. Transaminase Activity Predicts Survival in Patients with Head and Neck Cancer

    OpenAIRE

    Takenaka, Yukinori; Takemoto, Norihiko; Yasui, Toshimichi; Yamamoto, Yoshifumi; Uno, Atsuhiko; Miyabe, Haruka; Ashida, Naoki; Shimizu, Kotaro; Nakahara, Susumu; Hanamoto, Atshushi; Fukusumi, Takahito; Michiba, Takahiro; Cho, Hironori; Yamamoto, Masashi; Inohara, Hidenori

    2016-01-01

    Various serum biomarkers have been developed for predicting head and neck squamous cell carcinoma (HNSCC) prognosis. However, none of them have been proven to be clinically significant. A recent study reported that the ratio of aspartate aminotransaminase (AST) to alanine aminotransaminase (ALT) had a prognostic effect on non-metastatic cancers. This study aimed to examine the effect of the AST/ALT ratio on the survival of patients with HNSCC. Clinical data of 356 patients with locoregionally...

  5. Purification and preliminary crystallization of alanine racemase from Streptococcus pneumoniae

    Directory of Open Access Journals (Sweden)

    Im Hookang

    2007-05-01

    Full Text Available Abstract Background Over the past fifteen years, antibiotic resistance in the Gram-positive opportunistic human pathogen Streptococcus pneumoniae has significantly increased. Clinical isolates from patients with community-acquired pneumonia or otitis media often display resistance to two or more antibiotics. Given the need for new therapeutics, we intend to investigate enzymes of cell wall biosynthesis as novel drug targets. Alanine racemase, a ubiquitous enzyme among bacteria and absent in humans, provides the essential cell wall precursor, D-alanine, which forms part of the tetrapeptide crosslinking the peptidoglycan layer. Results The alanine racemases gene from S. pneumoniae (alrSP was amplified by PCR and cloned and expressed in Escherichia coli. The 367 amino acid, 39854 Da dimeric enzyme was purified to electrophoretic homogeneity and preliminary crystals were obtained. Racemic activity was demonstrated through complementation of an alr auxotroph of E. coli growing on L-alanine. In an alanine racemases photometric assay, specific activities of 87.0 and 84.8 U mg-1 were determined for the conversion of D- to L-alanine and L- to D-alanine, respectively. Conclusion We have isolated and characterized the alanine racemase gene from the opportunistic human pathogen S. pneumoniae. The enzyme shows sufficient homology with other alanine racemases to allow its integration into our ongoing structure-based drug design project.

  6. A methodology for cascade selection for co-product removal in the ω-transaminase system

    DEFF Research Database (Denmark)

    Janes, Kresimir; Gernaey, Krist; Tufvesson, Pär

    for choosing a feasible cascade system that will remove co-product to meet process requirements under process relevant conditions will be presented. Decisions are based on thermodynamic constraints, kinetics, selectivity, stability, pH change, cascade enzyme compatibility and downstream processing......Production of chiral amines using transaminases has indeed been proposed recently as an interesting alternative to conventional methods to help in the synthesis of many new pharmaceuticals. Two reaction strategies have been demonstrated: kinetic resolution and asymmetric synthesis. The latter...... pharmaceutical processes can only be achieved by selectively removing the product and/or co-product formed during the reaction (so called in-situ (co)product removal (IS(C)PR)). Several different alternative co-product removal strategies have been suggested, all of which have different impacts on the overall...

  7. Alanine aminotransferase controls seed dormancy in barley.

    Science.gov (United States)

    Sato, Kazuhiro; Yamane, Miki; Yamaji, Nami; Kanamori, Hiroyuki; Tagiri, Akemi; Schwerdt, Julian G; Fincher, Geoffrey B; Matsumoto, Takashi; Takeda, Kazuyoshi; Komatsuda, Takao

    2016-05-18

    Dormancy allows wild barley grains to survive dry summers in the Near East. After domestication, barley was selected for shorter dormancy periods. Here we isolate the major seed dormancy gene qsd1 from wild barley, which encodes an alanine aminotransferase (AlaAT). The seed dormancy gene is expressed specifically in the embryo. The AlaAT isoenzymes encoded by the long and short dormancy alleles differ in a single amino acid residue. The reduced dormancy allele Qsd1 evolved from barleys that were first domesticated in the southern Levant and had the long dormancy qsd1 allele that can be traced back to wild barleys. The reduced dormancy mutation likely contributed to the enhanced performance of barley in industrial applications such as beer and whisky production, which involve controlled germination. In contrast, the long dormancy allele might be used to control pre-harvest sprouting in higher rainfall areas to enhance global adaptation of barley.

  8. 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)

  9. 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.

  10. Immobilisation of ω-transaminase for industrial application: Screening and characterisation of commercial ready to use enzyme carriers

    DEFF Research Database (Denmark)

    Lima Afonso Neto, Watson; Schürmann, Martin; Panella, Lavinia

    2015-01-01

    Despite of the advantages that enzyme immobilisation can bring to industrial biocatalysis, its utilisation is still limited to a small number of enzymes and processes. Transaminase catalysed processes are a good example where immobilisation can be of major importance and even decisive for economic...

  11. Room-temperature solid phase ionic liquid (RTSPIL) coated Ω-transaminases: Development and application in organic solvents

    DEFF Research Database (Denmark)

    Grabner, B.; Nazario, M. A.; Gundersen, M. T.

    2018-01-01

    ω-Transaminases ATA-40, ATA-47 and ATA-82P were coated with room-temperature solid phase ionic liquids (RTSPILs) by means of three methods, melt coating, precipitation coating, and co‐lyophilization, and showed increased stability in all of the five tested organic solvents. Co‐lyophilization...

  12. Physiological and biochemical effects of morphactin IT 3233 on callus and tumour tissues of Nicotiana tabacum L. cultured in vitro III. Transamination processes catalysed by aminotransferase L-alanine: 2-oxoglutarate

    Directory of Open Access Journals (Sweden)

    Z. Chirek

    2015-01-01

    Full Text Available An active alanine transaminase was found both in callus and tumour tissues of tobacco. The enzyme is more active in the latter tissue, and the reaction balance is strongly shifted towards alanine production, while in callus tissue towards glutamic acid formation. Morphactin applied to the tissue cultures stimulates markedly the enzyme activity only in callus. A negative correlation was observed between the intensity of transamination processes and enhanced synthesis of proteins in the tissues studied. Morphactin disturbs nitrogen metabolism in the callus tissue. Tumour tissue is more resistant to the action of this substance. The different hormonal activities in these tissues may be the cause of the different effects of morphactin.

  13. 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

  14. Explaining Operational Instability of Amine Transaminases : Substrate-Induced Inactivation Mechanism and Influence of Quaternary Structure on Enzyme-Cofactor Intermediate Stability

    NARCIS (Netherlands)

    Borner, Tim; Ramisch, Sebastian; Reddem, Eswar R.; Bartsch, Sebastian; Vogel, Andreas; Thunnissen, Andy-Mark W. H.; Adlercreutz, Patrick; Grey, Carl

    The insufficient operational stability of amine transaminases (ATA) constitutes a limiting factor for high productivity in chiral amine synthesis. In this work, we investigated the operational stability of a tetrameric ATA with 92% sequence identity to a Pseudomonas sp. transaminase and compared it

  15. 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.

  16. Synchronization by Food Access Modifies the Daily Variations in Expression and Activity of Liver GABA Transaminase

    Directory of Open Access Journals (Sweden)

    Dalia De Ita-Pérez

    2014-01-01

    Full Text Available Daytime restricted feeding (DRF is an experimental protocol that influences the circadian timing system and underlies the expression of a biological clock known as the food entrained oscillator (FEO. Liver is the organ that reacts most rapidly to food restriction by adjusting the functional relationship between the molecular circadian clock and the metabolic networks. γ-Aminobutyric acid (GABA is a signaling molecule in the liver, and able to modulate the cell cycle and apoptosis. This study was aimed at characterizing the expression and activity of the mostly mitochondrial enzyme GABA transaminase (GABA-T during DRF/FEO expression. We found that DRF promotes a sustained increase of GABA-T in the liver homogenate and mitochondrial fraction throughout the entire day-night cycle. The higher amount of GABA-T promoted by DRF was not associated to changes in GABA-T mRNA or GABA-T activity. The GABA-T activity in the mitochondrial fraction even tended to decrease during the light period. We concluded that DRF influences the daily variations of GABA-T mRNA levels, stability, and catalytic activity of GABA-T. These data suggest that the liver GABAergic system responds to a metabolic challenge such as DRF and the concomitant appearance of the FEO.

  17. Magnetic catechol-chitosan with bioinspired adhesive surface: preparation and immobilization of ω-transaminase.

    Directory of Open Access Journals (Sweden)

    Kefeng Ni

    Full Text Available The magnetic chitosan nanocomposites have been studied intensively and been used practically in various biomedical and biological applications including enzyme immobilization. However, the loading capacity and the remained activity of immobilized enzyme based on existing approaches are not satisfied. Simpler and more effective immobilization strategies are needed. Here we report a simple catechol modified protocol for preparing a novel catechol-chitosan (CCS-iron oxide nanoparticles (IONPs composites carrying adhesive moieties with strong surface affinity. The ω-transaminase (ω-TA was immobilized onto this magnetic composite via nucleophilic reactions between catechol and ω-TA. Under optimal conditions, 87.5% of the available ω-TA was immobilized on the composite, yielding an enzyme loading capacity as high as 681.7 mg/g. Furthermore, the valuation of enzyme activity showed that ω-TA immobilized on CCS-IONPs displayed enhanced pH and thermal stability compared to free enzyme. Importantly, the immobilized ω-TA retained more than 50% of its initial activity after 15 repeated reaction cycles using magnetic separation and 61.5% of its initial activity after storage at 4°C in phosphate buffered saline (PBS for 15 days. The results suggested that such adhesive magnetic composites may provide an improved platform technology for bio-macromolecules immobilized.

  18. In vitro GABA-transaminase inhibitory compounds from the root of Angelica dahurica.

    Science.gov (United States)

    Choi, Soo Young; Ahn, Eun-Mi; Song, Myoung-Chong; Kim, Dae Won; Kang, Jeong Han; Kwon, Oh-Shin; Kang, Tae-Cheon; Baek, Nam-In

    2005-10-01

    The 80% aqueous MeOH extracts from the root of Angelica dahurica, found to inhibit the activities of GABA degradative enzymes GABA transaminase (GABA-T) and succinic semialdehyde dehydrogenase (SSADH), were fractionated using EtOAc, n-BuOH and H2O. Repeated column chromatography for the EtOAc and n-BuOH fractions led to the isolation of two new coumarins, oxypeucedanin hydrate-3''-butyl ether and isopraeroside IV along with six known coumarins, isoimperatorin, imperatorin, phellopterin, oxypeucedanin hydrate, nodakenin and 3'-hydroxymarmesinin, and two polyacetylenes, falcarindiol and octadeca-1,9-dien-4,6-diyn-3,8,18-triol. Of the isolated pure compounds, imperatorin and falcarindiol inactivated the GABA-T activities in both time- and concentration-dependent manners. The kinetic studies showed that imperatorin and falcarindiol reacted with the GABA-T with a second-order rate constant of 2.3 +/- 0.2 mm(-1) min(-1) and 1.5 +/- 0.1 mm(-1) min(-1), respectively. It is postulated that imperatorin and falcarindiol are able to elevate the neurotransmitter GABA levels in the central nervous system by an inhibitory action on the GABA degradative enzyme GABA-T. Copyright (c) 2005 John Wiley & Sons, Ltd.

  19. Identification of novel transaminases from a 12-aminododecanoic acid-metabolizing Pseudomonas strain.

    Science.gov (United States)

    Wilding, Matthew; Walsh, Ellen F A; Dorrian, Susan J; Scott, Colin

    2015-07-01

    A Pseudomonas species [Pseudomonas sp. strain amino alkanoate catabolism (AAC)] was identified that has the capacity to use 12-aminododecanoic acid, the constituent building block of homo-nylon-12, as a sole nitrogen source. Growth of Pseudomonas sp. strain AAC could also be supported using a range of additional ω-amino alkanoates. This metabolic function was shown to be most probably dependent upon one or more transaminases (TAs). Fourteen genes encoding putative TAs were identified from the genome of Pseudomonas sp. AAC. Each of the 14 genes was cloned, 11 of which were successfully expressed in Escherichia coli and tested for activity against 12-aminododecanoic acid. In addition, physiological functions were proposed for 9 of the 14 TAs. Of the 14 proteins, activity was demonstrated in 9, and of note, 3 TAs were shown to be able to catalyse the transfer of the ω-amine from 12-aminododecanoic acid to pyruvate. Based on this study, three enzymes have been identified that are promising biocatalysts for the production of nylon and related polymers. © 2015 The Authors. Microbial Biotechnology published by John Wiley & Sons Ltd and Society for Applied Microbiology.

  20. [The role of transaminases and mitochondrial anion porters in the energetics of animals of different ages].

    Science.gov (United States)

    Akhmerov, R N; Sultanov, Sh; Allamuratov, Sh I

    1995-01-01

    Aminohydroxyacetate, an inhibitor of aminotransferases, decreases the rate of oxygen consumption by 1-day-old young rats by 35% 10 min after intraperitoneal injection, whereas for 20-day-old rats, the inhibitory effect is 56%, and for adult mice, it is 83%. More prolonged exposure to aminohydroxyacetate leads to death of the animal. One-day-old rats die 90 min after the injection, 20-day-old rats die 30 min after the injection, and adult mice die 15 min after the injection. Butylmalonate (an inhibitor of mitochondrial dicarboxylate translocator) decreases the rate of energy metabolism to a lower extent, by 38% in adult mice and by 18% in 1-day-old rats. The animals generally remain alive after the exposure to this compound. 1,2,3-benzyltricarboxylate, an inhibitor of tricarboxylate transport, shows only a weak effect on energy metabolism of animals of any age. These results provide evidence that the role of the transaminase system in energy metabolism increases with age. Mechanisms underlying weak sensitivity of newborn animals to these inhibitors are discussed.

  1. Immobilization of ω-transaminase by magnetic PVA-Fe3O4nanoparticles.

    Science.gov (United States)

    Jia, Honghua; Huang, Fan; Gao, Zhen; Zhong, Chao; Zhou, Hua; Jiang, Min; Wei, Ping

    2016-06-01

    ω-Transaminase (ω-TA) as a kind of important biocatalyst is widely used in preparation of chiral intermediates. In this paper, a magnetic PVA-Fe 3 O 4 nanoparticles was prepared and employed on immobilization of ω-TA to reduce the cost, increase reusability and enhance stability. The prepared magnetic PVA-Fe 3 O 4 nanoparticles were characterized by transmission electron microscope (TEM), X-ray diffraction (XRD) and vibrating sample magnetometer (VSM). The average size of magnetic PVA-Fe 3 O 4 nanoparticles was located between 30-40 nm ω-TA was immobilized onto magnetic PVA-Fe 3 O 4 nanoparticles via glutaraldehyde cross-linking, achieving a wider pH range as 6-8 and also a better thermal stability at 60 °C. Meanwhile, the immobilized ω-TA could be successfully reused for 13 times in biotransformation. These results therefore indicated that the immobilized ω-TA with high stability might be readily utilized in industrial purposes.

  2. 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%.

  3. β-Alanine supplementation for athletic performance: an update.

    Science.gov (United States)

    Bellinger, Phillip M

    2014-06-01

    β-alanine supplementation has become a common practice among competitive athletes participating in a range of different sports. Although the mechanism by which chronic β-alanine supplementation could have an ergogenic effect is widely debated, the popular view is that β-alanine supplementation augments intramuscular carnosine content, leading to an increase in muscle buffer capacity, a delay in the onset of muscular fatigue, and a facilitated recovery during repeated bouts of high-intensity exercise. β-alanine supplementation appears to be most effective for exercise tasks that rely heavily on ATP synthesis from anaerobic glycolysis. However, research investigating its efficacy as an ergogenic aid remains equivocal, making it difficult to draw conclusions as to its effectiveness for training and competition. The aim of this review was to update, summarize, and critically evaluate the findings associated with β-alanine supplementation and exercise performance with the most recent research available to allow the development of practical recommendations for coaches and athletes. A critical review of the literature reveals that when significant ergogenic effects have been found, they have been generally shown in untrained individuals performing exercise bouts under laboratory conditions. The body of scientific data available concerning highly trained athletes performing single competition-like exercise tasks indicates that this type of population receives modest but potentially worthwhile performance benefits from β-alanine supplementation. Recent data indicate that athletes may not only be using β-alanine supplementation to enhance sports performance but also as a training aid to augment bouts of high-intensity training. β-alanine supplementation has also been shown to increase resistance training performance and training volume in team-sport athletes, which may allow for greater overload and superior adaptations compared with training alone. The ergogenic

  4. The antiproton depth–dose curve measured with alanine detectors

    CERN Document Server

    Bassler, Niels; Palmans, Hugo; Holzscheiter, Michael H; Kovacevic, Sandra

    2008-01-01

    n this paper we report on the measurement of the antiproton depth–dose curve, with alanine detectors. The results are compared with simulations using the particle energy spectrum calculated by FLUKA, and using the track structure model of Hansen and Olsen for conversion of calculated dose into response. A good agreement is observed between the measured and calculated relative effectiveness although an underestimation of the measured values beyond the Bragg-peak remains unexplained. The model prediction of response of alanine towards heavy charged particles encourages future use of the alanine detectors for dosimetry of mixed radiation fields.

  5. The Antiproton Depth Dose Curve Measured with Alanine Detectors

    DEFF Research Database (Denmark)

    Bassler, Niels; Hansen, Johnny Witterseh; Palmans, Hugo

    2008-01-01

    In this paper we report on the measurement of the antiproton depth dose curve, with alanine detectors. The results are compared with simulations using the particle energy spectrum calculated by FLUKA, and using the track structure model of Hansen et Olsen for conversion of calculated dose...... into response. A good agreement was observed between the measured and calculated relative effectiveness although a slight underestimation of the calculated values in the Bragg peak remains unexplained. The model prediction of response of alanine towards heavy charged particles encourages future use...... of the alanine detectors for dosimetry of mixed radiation fields....

  6. 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)

  7. 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)

  8. 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.

  9. Crystallization and preliminary X-ray diffraction analysis of ω-amino acid:pyruvate transaminase from Chromobacterium violaceum

    International Nuclear Information System (INIS)

    Sayer, Christopher; Isupov, Michail N.; Littlechild, Jennifer A.

    2007-01-01

    An ω-amino acid:pyruvate transaminase from C. violaceum has been purified and crystallized in two crystal forms. The structure has been solved using molecular replacement. The enzyme ω-transaminase catalyses the conversion of chiral ω-amines to ketones. The recombinant enzyme from Chromobacterium violaceum has been purified to homogeneity. The enzyme was crystallized from PEG 4000 using the microbatch method. Data were collected to 1.7 Å resolution from a crystal belonging to the triclinic space group P1, with unit-cell parameters a = 58.9, b = 61.9, c = 63.9 Å, α = 71.9, β = 87.0, γ = 74.6°. Data were also collected to 1.95 Å from a second triclinic crystal form. The structure has been solved using the molecular-replacement method

  10. 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)

  11. Alanine administration does not stimulate gluconeogenesis in preterm infants

    NARCIS (Netherlands)

    van Kempen, Anne A. M. W.; Romijn, Johannes A.; Ruiter, An F. C.; Endert, Erik; Weverling, Gerrit Jan; Kok, Johanna H.; Sauerwein, Hans P.

    2003-01-01

    Gluconeogenesis partially depends on sufficient precursor supply, and plasma alanine concentrations are generally low in preterm infants. Stimulation of gluconeogenesis may contribute to the prevention of hypoglycemia, an important clinical problem in these infants. In this study we evaluated the

  12. A High Sensitivity EPR Technique for Alanine Dosimetry (invited paper)

    International Nuclear Information System (INIS)

    Haskell, E.H.; Hayes, R.B.; Kenner, G.H.

    1998-01-01

    Uncertainties of ± 5 mGy were achieved in the measurement of alanine dosemeters using optimised EPR parameters, instrumentation, spectral manipulation and subtraction techniques. Modulation amplitude and microwave power were adjusted to combine resonances of two neighbouring alanine signals. Instrumental variations were minimised by combining and subtracting pre- and post-measurement spectra of the empty EPR tube. A spectrum of the native signal of non-dosed alanine was generated from a single batch of dosemeters and subtracted from spectra of the irradiated dosemeters, also from the same batch. Field alignment was adjusted with the use of an in-cavity Mn ++ standard. A constant rotation goniometer was used to eliminate anisotropies in the EPR tube and alanine samples. Finally, digital filters were applied to the resulting spectra. (author)

  13. 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)

  14. 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)

  15. 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

  16. Kinetic resolution of chiral amines with omega-transaminase using an enzyme-membrane reactor.

    Science.gov (United States)

    Shin, J S; Kim, B G; Liese, A; Wandrey, C

    2001-05-05

    A kinetic resolution process for the production of chiral amines was developed using an enzyme-membrane reactor (EMR) and a hollow-fiber membrane contactor with (S)-specific omega-transaminases (omega-TA) from Vibrio fluvialis JS17 and Bacillus thuringiensis JS64. The substrate solution containing racemic amine and pyruvate was recirculated through the EMR and inhibitory ketone product was selectively extracted by the membrane contactor until enantiomeric excess of (R)-amine exceeded 95%. Using the reactor set-up with flat membrane reactor (10-mL working volume), kinetic resolutions of alpha-methylbenzylamine (alpha-MBA) and 1-aminotetralin (200 mM, 50 mL) were carried out. During the operation, concentration of ketone product, i.e., acetophenone or alpha-tetralone, in a substrate reservoir was maintained below 0.1 mM, suggesting efficient removal of the inhibitory ketone by the membrane contactor. After 47 and 32.5 h of operation using 5 U/mL of enzyme, 98.0 and 95.5% ee of (R)-alpha-MBA and (R)-1-aminotetralin were obtained at 49.5 and 48.8% of conversion, respectively. A hollow-fiber membrane reactor (39-mL working volume) was used for a preparative-scale kinetic resolution of 1-aminotetralin (200 mM, 1 L). After 133 h of operation, enantiomeric excess reached 95.6% and 14.3 g of (R)-1-aminotetralin was recovered (97.4% of yield). Mathematical modeling of the EMR process including the membrane contactor was performed to evaluate the effect of residence time. The simulation results suggest that residence time should be short to maintain the concentration of the ketone product in EMR sufficiently low so as to decrease conversion per cycle and, in turn, reduce the inhibition of the omega-TA activity. Copyright 2001 John Wiley & Sons, Inc.

  17. Hepatic transaminase and alkaline phosphatase enzyme levels in HIV/HBV co-infected and HIV mono-infected patients in Maiduguri, Nigeria.

    Science.gov (United States)

    Goni, B W; Yusuph, H; Mustapha, S K; Sahabi, M A; Gwalabe, S A; Tahir, A; Bakki, B; Kida, I M

    2013-01-01

    Studies have shown that HIV-HBV co-infected patients have an increased risk of liver-related morbidity and mortality compared to their HIV-mono-infected counterparts. Furthermore, it has been reported that HIV-HBV co-infected patients have a significantly high incidence of drug-induced hepatotoxicity following commencement of HAART than HIV-mono-infected patients. To compare the levels of aspartate amino transferase (AST), alanine amino transferase (ALT) and alkaline phosphatase (ALKPO 4 ) enzyme levels between HAART naïve HIV-HBV co-infected patients and their HIV-mono-infected counterparts. A cross-sectional descriptive study in which 142 newly diagnosed HIV/HBV co-infected and HIV mono-infected adults were investigated for alkaline aminotransferase, aspartate aminotransferase and alkaline phosphatase enzyme levels. The study subjects comprised of 80 (56.3%) females and 62 (46.7%) males. The age range of the study population was 15-65 years. The mean ages of male and female subjects were 45.5 ± 10.5 years and 39.1 ± 7.5 years respectively ( P mono-infected. The mean ALT enzyme level of HIV/HBV co-infected subjects was significantly higher than that of HIV mono-infected ones i.e., 42.12 IU/l vs. 27.86 IU/l, ( P = 0.038). However, there was no statistically significant difference in the mean AST (30.14 IU/l vs. 29.09 IU/l, P = 0.893) and ALKPO 4 (55.86 IU/l vs. 60.97 IU/l, P = 0.205) enzyme levels between HIV-HBV co-infected and HIV mono-infected subjects albeit the two enzymes were moderately elevated in both categories of subjects. The significantly elevated ALT enzyme levels amongst HIV-HBV co-infected subjects suggest that HIV-HBV co-infected patients may have an increased risk of liver-related morbidity and mortality than their HIV mono-infected counterparts. Screening for serological markers of chronic HBV infection, as well as hepatic transaminase enzyme levels in all newly diagnosed HIV-positive patients is therefore recommended before commencement of

  18. Determination of Alanine concentration in Alanine-supercritical water mixture, using UNIQUAC methods and density functional theory

    Directory of Open Access Journals (Sweden)

    Vahid mohammadrezaee

    2016-12-01

    Full Text Available In the following paper, density functional method and Universal QUAsi-chemical (UNIQUA activity index model were applied to calculate the concentration of Alanine I supercritical water. B3LYB method and 6-311G+ (2D, 2P basic function were used to optimize the molecular structure of Alanine, water and Alanine water complex. Then, reaction parameters of UNIQUAC model were calculated, using formation energy values. PCM method and zeropoint energy were utilized to include solvent effect and BSSE effect respectively. The resulted errors indicate that the method in a fairly good agreement with the experimental data. Including SSE in the interaction parameter calculations reduces the total error significantly. According to our calculations theAlanine- water complex showed a negative deviation from Raoul’s law.

  19. Place des alanine-aminotransferases dans la securite ...

    African Journals Online (AJOL)

    The serological assays' performance seems to deny all interest to the ALAT determination in the blood safety. Nous avons mené une étude sur 164 donneurs de sang bénévoles au Centre National de Transfusion Sanguine (CNTS) de Lomé pour évaluer la place de la mesure de l'activité des transaminases de type

  20. Alanine turnover in the postabsorptive state and during parenteral hyperalimentation before and after surgery

    NARCIS (Netherlands)

    Sauerwein, H. P.; Michels, R. P.; Cejka, V.

    1981-01-01

    Influence of total parenteral nutrition and operation on alanine turnover and venous alanine concentration was determined in 5 patients with stomach carcinoma using single technique of U-14C alanine. Every patient served at his own control. In the postabsorptive state alanine turnover was 1.63 +/-

  1. 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.

  2. Control of alanine metabolism in rat liver by transport processes or cellular metabolism.

    OpenAIRE

    Fafournoux, P; Rémésy, C; Demigné, C

    1983-01-01

    1. Factors governing hepatic utilization of alanine were studied in vivo and in vitro in rats adapted to increasing dietary protein. 2. Hepatic alanine utilization was enhanced 5-fold with a 90%-casein diet, compared with a 13%-casein diet. The increased uptake resulted from enhanced fractional extraction in the presence of high concentrations of alanine in the portal vein. 3. The increase in alanine metabolism on high-protein diets was associated with an increase in alanine aminotransferase ...

  3. Caramelization of maltose solution in presence of alanine.

    Science.gov (United States)

    Fadel, H H M; Farouk, A

    2002-01-01

    Two solutions of maltose in water were used to prepare caramels. Alanine as a catalyst was added to one of these solutions. The caramelization was conducted at 130 degrees C for total time period 90 minutes. Convenient samples were taken of each caramel solution every 30 min and subjected to sensory analysis and isolation of volatile components. The odour and colour sensory tests were evaluated according to the international standard methods (ISO). The results showed that, the presence of alanine gave rise to a high significant (P caramel attributes, which are the most important caramel notes. On the other hand the increase in heating time in presence of alanine as a catalyst resulted in a high significant (P caramel solution. The new technique Solid Phase Micro Extraction (SPME) was used for trapping the volatile components in the headspace of each caramel samples followed by thermal desorption and GC and GC - MS analysis. The 5-hydroxymethyl-2-furfural (HMF), the main characteristic caramel product, showed its highest value in sample containing alanine after heating for 60 minutes. The best sensory results of the sample contains alanine were confirmed by the presence of high concentrations of the most potent odorants of caramel besides to the formation of some volatile compounds have caramel like flavours such as 2-acetyl pyrrole, 2-furanones and 1-(2-furanyl)1,2-propandione.

  4. 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...

  5. EPR structure of the gamma irradiated alanine spectrum

    International Nuclear Information System (INIS)

    Cabral P, A.; Jimenez D, H.; Urena N, F.; Galindo, S.; Bosch, P.

    1992-03-01

    In this study is shown that the broadened five-line EPR pattern of the gamma irradiated alanine possibly decomposes into a more complex pattern when the recorded spectrum is subject to an operation of deconvolution. The EPR powder spectra of gamma irradiated DL- and L-alanine with and without binders are analysed. In all recorded spectra, each observed line is resolved into an asymmetrical triplet when a Gaussian distribution of 8.2 gauss width is removed, by deconvolution, from the observed spectrum. On the other hand, from a simple fitting analysis carried out on the original data, one encounters that some calculated relations between characteristic parameters, such as intensity ratios, deviate consistently from assumed height ratios. Both, from deconvolution and fitting results, a different structure is suggested for the observed broadened five-line EPR pattern of γ-irradiated powder DL- and L-alanine. (Author)

  6. Structure and Intramolecular Proton Transfer of Alanine Radical Cations

    International Nuclear Information System (INIS)

    Lee, Gab Yong

    2012-01-01

    The structures of the four lowest alanine conformers, along with their radical cations and the effect of ionization on the intramolecular proton transfer process, are studied using the density functional theory and MP2 method. The energy order of the radical cations of alanine differs from that of the corresponding neutral conformers due to changes in the basicity of the NH 2 group upon ionization. Ionization favors the intramolecular proton transfer process, leading to a proton-transferred radical-cation structure, [NH 3 + -CHCH 3 -COO·], which contrasts with the fact that a proton-transferred zwitterionic conformer is not stable for a neutral alanine in the gas phase. The energy barrier during the proton transfer process is calculated to be about 6 kcal/mol

  7. EPR structure of the gamma irradiated alanine spectrum

    Energy Technology Data Exchange (ETDEWEB)

    Cabral P, A.; Jimenez D, H.; Urena N, F.; Galindo, S.; Bosch, P

    1992-03-15

    In this study is shown that the broadened five-line EPR pattern of the gamma irradiated alanine possibly decomposes into a more complex pattern when the recorded spectrum is subject to an operation of deconvolution. The EPR powder spectra of gamma irradiated DL- and L-alanine with and without binders are analysed. In all recorded spectra, each observed line is resolved into an asymmetrical triplet when a Gaussian distribution of 8.2 gauss width is removed, by deconvolution, from the observed spectrum. On the other hand, from a simple fitting analysis carried out on the original data, one encounters that some calculated relations between characteristic parameters, such as intensity ratios, deviate consistently from assumed height ratios. Both, from deconvolution and fitting results, a different structure is suggested for the observed broadened five-line EPR pattern of {gamma}-irradiated powder DL- and L-alanine. (Author)

  8. Effects of dose fractionation on the response of alanine dosimetry

    International Nuclear Information System (INIS)

    Lundahl, Brad; Logar, John; Desrosiers, Marc; Puhl, James

    2014-01-01

    Alanine dosimetry is well established as a transfer standard and is becoming more prevalently used in routine dosimetry systems for radiation processing. Many routine measurement applications in radiation processing involve absorbed dose measurements resulting from fractioned exposures to ionizing radiation. Fractioning of absorbed dose is identified as an influence quantity (ISO/ASTM, 2013). This paper reports on study results of absorbed dose fractioning characteristics of alanine for gamma and high energy electron beam radiation sources. The results of this study indicate a radiation response difference due to absorbed dose fractioning in response can be observed after four fractionations for high-energy electron beams and no difference up to seven fractions for gamma rays using an ANOVA evaluation method. - Highlights: • Fractioning effects signaled in electron beam using an ANOVA at 6 equal increments. • Fractioning effects not signaled in gamma using an ANOVA up to 7 equal increments. • Insensitivity of alanine to dose fractioning indicates nominal impact on calibration

  9. Propionate enters GABAergic neurons, inhibits GABA transaminase, causes GABA accumulation and lethargy in a model of propionic acidemia.

    Science.gov (United States)

    Morland, Cecilie; Frøland, Anne-Sofie; Pettersen, Mi Nguyen; Storm-Mathisen, Jon; Gundersen, Vidar; Rise, Frode; Hassel, Bjørnar

    2018-02-16

    Propionic acidemia is the accumulation of propionate in blood due to dysfunction of propionyl-CoA carboxylase. The condition causes lethargy and striatal degeneration with motor impairment in humans. How propionate exerts its toxic effect is unclear. Here, we show that intravenous administration of propionate causes dose-dependent propionate accumulation in the brain and transient lethargy in mice. Propionate, an inhibitor of histone deacetylase, entered GABAergic neurons, as could be seen from increased neuronal histone H4 acetylation in the striatum and neocortex. Propionate caused an increase in GABA (γ-amino butyric acid) levels in the brain, suggesting inhibition of GABA breakdown. In vitro propionate inhibited GABA transaminase with a K i of ∼1 mmol/l. In isolated nerve endings, propionate caused increased release of GABA to the extracellular fluid. In vivo , propionate reduced cerebral glucose metabolism in both striatum and neocortex. We conclude that propionate-induced inhibition of GABA transaminase causes accumulation of GABA in the brain, leading to increased extracellular GABA concentration, which inhibits neuronal activity and causes lethargy. Propionate-mediated inhibition of neuronal GABA transaminase, an enzyme of the inner mitochondrial membrane, indicates entry of propionate into neuronal mitochondria. However, previous work has shown that neurons are unable to metabolize propionate oxidatively, leading us to conclude that propionyl-CoA synthetase is probably absent from neuronal mitochondria. Propionate-induced inhibition of energy metabolism in GABAergic neurons may render the striatum, in which >90% of the neurons are GABAergic, particularly vulnerable to degeneration in propionic acidemia. © 2018 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.

  10. Transaminase Activity Predicts Survival in Patients with Head and Neck Cancer.

    Science.gov (United States)

    Takenaka, Yukinori; Takemoto, Norihiko; Yasui, Toshimichi; Yamamoto, Yoshifumi; Uno, Atsuhiko; Miyabe, Haruka; Ashida, Naoki; Shimizu, Kotaro; Nakahara, Susumu; Hanamoto, Atshushi; Fukusumi, Takahito; Michiba, Takahiro; Cho, Hironori; Yamamoto, Masashi; Inohara, Hidenori

    2016-01-01

    Various serum biomarkers have been developed for predicting head and neck squamous cell carcinoma (HNSCC) prognosis. However, none of them have been proven to be clinically significant. A recent study reported that the ratio of aspartate aminotransaminase (AST) to alanine aminotransaminase (ALT) had a prognostic effect on non-metastatic cancers. This study aimed to examine the effect of the AST/ALT ratio on the survival of patients with HNSCC. Clinical data of 356 patients with locoregionally advanced HNSCC were collected. The effect of the AST/ALT ratio on overall survival was analyzed using a Cox proportional hazard model. Moreover, recursive partitioning analysis (RPA) was used to divide the patients into groups on the basis of the clinical stage and AST/ALT ratio. The prognostic ability of this grouping was validated using an independent data set (N = 167). The AST/ALT ratio ranged from 0.42 to 4.30 (median, 1.42) and was a prognostic factor for overall survival that was independent of age, primary sites, and tumor stage (hazard ratio: 1.36, confidence interval: 1.08-1.68, P = 0.010). RPA divided patients with stage IVA into the following two subgroups: high AST/ALT (≥2.3) and low AST/ALT (AST/ALT ratio, stage IVA with a high AST/ALT ratio, and stage IVB were 64.8%, 49.2%, 28.6%, and 33.3%, respectively (p AST/ALT group, the adjusted hazard ratio for death was 2.17 for high AST/ALT group (confidence interval: 1.02-.22 P = 0.045). The AST/ALT ratio was demonstrated to be a prognostic factor of HNSCC. The ratio subdivided patients with stage IVA into low- and high-risk groups. Moreover, intensified treatment for the high-risk group may be considered.

  11. The features of radiation damages in L-alanine crystals

    International Nuclear Information System (INIS)

    Zaitov, V.R.; Onischuk, V.A.

    1996-01-01

    The method of the ESR alanine dosimetry has appeared the most convenient one for measurement of radiation dose in the range 1-10 6 Gy. Its peculiarities are the wide dose range, the high accuracy, the absence fading at room temperature, the possibility of many times repeated measurements as dosemeter accumulates dose, the simplicity of measurements. Because of this performance ESR alanine dosimetry technique can be applied to continuous monitoring radiation doses absorbed by materials on nuclear power stations as well as of dose fields and restoration doses after an accident situation. In order to determine accurately the absorbed dose in an accident on background of accumulated dose for previous period, it is necessary to the utmost increase the accuracy of dosimetry system. For this reason it is necessary to know how the properties of free radicals which formings in irradiated L-alanine are displayed in signal ESR. With the purpose to detect the structure of the free radicals the ESR spectra the L-alanine and L-alanine-d 3 single crystals were studied. The samples were grown by slow evaporation of the saturated aqueous solution. For obtain the L-alanine-d 3 the three-divisible recrystallization in heavy water had been used. The samples were irradiated with 60 Co at room temperature and in liquid nitrogen. The irradiation doses were 10 kGy and dose rate was 8,3 Gy/s. To increase the resolution of the ESR spectra hyperfine structure the second derivative for the absorption curve was registered. The measurements were conducted in X-range at temperatures 77-430 K. (author)

  12. Medical reference dosimetry using EPR measurements of alanine

    DEFF Research Database (Denmark)

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

    2009-01-01

    signal fading, non-destructive measurement and small dosimeter size. Material and Methods. A Bruker EMX-micro EPR spectrometer with a rectangular cavity and a measurement time of two minutes per dosimeter was used for reading of irradiated alanine dosimeters. Under these conditions a new algorithm based...... 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....

  13. Ab initio study of alanine polypeptide chain twisting

    DEFF Research Database (Denmark)

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

    2006-01-01

    We have investigated the potential energy surfaces for alanine chains consisting of three and six amino acids. For these molecules we have calculated potential energy surfaces as a function of the Ramachandran angles ph$ and psi, which are widely used for the characterization of the polypeptide...... and with the available experimental data extracted from the Protein Data Base. This comparison demonstrates a reasonable correspondence of the most prominent minima on the calculated potential energy surfaces to the experimentally measured angles phi and psi for alanine chains appearing in native proteins. We have also...

  14. Stereoselective aminoacylation of a dinucleoside monophosphate by the imidazolides of DL-alanine and N-(tert-butoxycarbonyl)-DL-alanine

    Science.gov (United States)

    Profy, A. T.; Usher, D. A.

    1984-01-01

    The aminoacylation of diinosine monophosphate was studied experimentally. When the acylating agent was the imidazolide of N-(tert-butoxycarbonyl)-DL-alanine, a 40 percent enantiomeric excess of the isomer was incorporated at the 2' site and the positions of equilibrium for the reversible 2'-3' migration reaction differed for the D and L enantiomers. The reactivity of the nucleoside hydroxyl groups was found to decrease on the order 2'(3') less than internal 2' and less than 5', and the extent of the reaction was affected by the concentration of the imidazole buffer. Reaction of IpI with imidazolide of unprotected DL-alanine, by contrast, led to an excess of the D isomer at the internal 2' site. Finally, reaction with the N-carboxy anhydride of DL-alanine occurred without stereoselection. These results are found to be relevant to the study of the evolution of optical chemical activity and the origin of genetically directed protein synthesis.

  15. Supported liquid membrane as a novel tool for driving the equilibrium of ω-transaminase catalyzed asymmetric synthesis.

    Science.gov (United States)

    Rehn, Gustav; Adlercreutz, Patrick; Grey, Carl

    2014-06-10

    An attractive option to produce chiral amines of industrial importance is through asymmetric synthesis using ω-transaminase. However, reaching high yields often requires a strategy for shifting the equilibrium position. This paper describes a novel strategy for handling this problem. It involves the use of a supported liquid membrane (SLM) together with a packed bed reactor. The reactor contains Escherichia coli cells with ω-transaminase from Arthrobacter citreus, immobilized by flocculation with chitosan. The SLM consists of a hollow fibre membrane contactor in which the pores contain undecane. The system enables continuous extraction of the amine product and was used to successfully shift the equilibrium in asymmetric synthesis of (S)-α-methylbenzylamine (MBA). A conversion of 98% was reached, compared to 50% without product extraction. Moreover, a selective extraction of the produced MBA was realized. A high product concentration of 55g/l was reached after 80h, and the system showed promising potential for continuous operation. Copyright © 2014 Elsevier B.V. All rights reserved.

  16. 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...

  17. Correlation between alanine aminotransferase level, HCV-RNA titer ...

    African Journals Online (AJOL)

    Reham Al Swaff

    2012-04-04

    Apr 4, 2012 ... Abstract The relationship of serum alanine aminotransferase (ALT) level and viral replication to liver damage in chronic hepatitis C virus (HCV) patients remains unclear. The aim of the present study was to determine whether the stage of fibrosis correlates with HCV-. RNA titer and/or serum ALT level in ...

  18. Pressure-induced phase transformations in L-alanine crystals

    DEFF Research Database (Denmark)

    Olsen, J. Staun; Gerward, Leif; Freire, P.T.C.

    2008-01-01

    Raman scattering and synchrotron X-ray diffraction have been used to investigate the high-pressure behavior of L-alanine. This study has confirmed a structural phase transition observed by Raman scattering at 2.3 GPa and identified it as a change from orthorhombic to tetragonal structure. Another...

  19. 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.

  20. Thin layer alanine dosimeter with optical spectrophotometric evaluation

    International Nuclear Information System (INIS)

    Zagorski, Z.P.

    2000-01-01

    Experience in the high dose dosimetry of gamma radiation, gathered in our group from the sixties till now, allows to express the opinion, that techniques applied are adequate to solve problems. It can be confirmed by the fact that 60% of laboratories participating in the international comparison during the duration of the contract obtained satisfactory results. Adaptation of these methods, in particular of the alanine-ESR dosimetry to highly inhomogeneous fields of EB gives poor results, as it has been shown on thin films of the alanine/polymer composite. However, the applications of these films give excellent results if the concentration of the radical CH 3 C·H CO 2 - is measured by diffuse reflection spectrophotometry, which tolerates poor transparency of the composite and is insensitive to the orientation of crystals of alanine in thin films, what is disqualifying the ESR measurements. The development of thin-film dosimeters for EB processing was possible due to new developments in solid state radiation chemistry. The research has revealed some unsolved questions, e.g. of the high temperature coefficient of alanine based dosimeters, of the role of the size of spurs and the necessity to adapt dosimetry to the energy spectrum of electrons, because every type of accelerators differs in that respect. (author)

  1. 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

  2. Microhardness studies on nonlinear optical L-alanine single crystals

    Indian Academy of Sciences (India)

    Sci., Vol. 36, No. 3, June 2013, pp. 471–474. c Indian Academy of Sciences. Microhardness studies on nonlinear optical L-alanine single crystals. R HANUMANTHARAO† and S KALAINATHAN‡,∗ ... ter to the area of the impression left on the specimen. Both ... where P is the applied load in kg, d is in mm and Hv is in kg mm.

  3. Impact of weight reduction program on serum alanine ...

    African Journals Online (AJOL)

    Conclusion: Based on our findings, weight loss modulates serum alanine aminotransferase and immune system parameters of patients with hepatitis B virus infection. Keywords: Hepatitis B virus, obesity, immune system, weight reduction. DOI: http://dx.doi.org/10.4314/ahs.v16i1.17. Cita as: Abd El-Kader SM, Al-Dahr MHS.

  4. The unresolved puzzle why alanine extensions cause disease.

    Science.gov (United States)

    Winter, Reno; Liebold, Jens; Schwarz, Elisabeth

    2013-08-01

    The prospective increase in life expectancy will be accompanied by a rise in the number of elderly people who suffer from ill health caused by old age. Many diseases caused by aging are protein misfolding diseases. The molecular mechanisms underlying these disorders receive constant scientific interest. In addition to old age, mutations also cause congenital protein misfolding disorders. Chorea Huntington, one of the most well-known examples, is caused by triplet extensions that can lead to more than 100 glutamines in the N-terminal region of huntingtin, accompanied by huntingtin aggregation. So far, nine disease-associated triplet extensions have also been described for alanine codons. The extensions lead primarily to skeletal malformations. Eight of these proteins represent transcription factors, while the nuclear poly-adenylate binding protein 1, PABPN1, is an RNA binding protein. Additional alanines in PABPN1 lead to the disease oculopharyngeal muscular dystrophy (OPMD). The alanine extension affects the N-terminal domain of the protein, which has been shown to lack tertiary contacts. Biochemical analyses of the N-terminal domain revealed an alanine-dependent fibril formation. However, fibril formation of full-length protein did not recapitulate the findings of the N-terminal domain. Fibril formation of intact PABPN1 was independent of the alanine segment, and the fibrils displayed biochemical properties that were completely different from those of the N-terminal domain. Although intranuclear inclusions have been shown to represent the histochemical hallmark of OPMD, their role in pathogenesis is currently unclear. Several cell culture and animal models have been generated to study the molecular processes involved in OPMD. These studies revealed a number of promising future therapeutic strategies that could one day improve the quality of life for the patients.

  5. Elevated transaminases as a predictor of coma in a patient with anorexia nervosa: a case report and review of the literature

    Directory of Open Access Journals (Sweden)

    Yoshida Shuhei

    2010-09-01

    Full Text Available Abstract Introduction Liver injury is a frequent complication associated with anorexia nervosa, and steatosis of the liver is thought to be the major underlying pathology. However, acute hepatic failure with transaminase levels over 1000 IU/mL and deep coma are very rare complications and the mechanism of pathogenesis is largely unknown. Case presentation A 37-year-old Japanese woman showed features of acute liver failure and hepatic coma which were not associated with hypoglycemia or hyper-ammonemia. Our patient's consciousness was significantly improved with the recovery of liver function and normalization of transaminase levels after administration of nutritional support. Conclusions Our case report demonstrates that transaminase levels had an inverse relationship with the consciousness of our patient, although the pathogenesis of coma remains largely unknown. This indicates that transaminase levels can be one of the key predictors of impending coma in patients with anorexia nervosa. Therefore, frequent monitoring of transaminase levels combined with rigorous treatment of the underlying nutritional deficiency and psychiatric disorder are necessary to prevent this severe complication.

  6. The substrate specificity, enantioselectivity and structure of the (R)-selective amine : pyruvate transaminase from Nectria haematococca

    Science.gov (United States)

    Sayer, Christopher; Martinez-Torres, Ruben J; Richter, Nina; Isupov, Michail N; Hailes, Helen C; Littlechild, Jennifer A; Ward, John M

    2014-01-01

    During the last decade the use of transaminases for the production of pharmaceutical and fine chemical intermediates has attracted a great deal of attention. Transaminases are versatile biocatalysts for the efficient production of amine intermediates and many have (S)-enantiospecificity. Transaminases with (R)-specificity are needed to expand the applications of these enzymes in biocatalysis. In this work we have identified a fungal putative (R)-specific transaminase from the Eurotiomycetes Nectria haematococca, cloned a synthetic version of this gene, demonstrated (R)-selective deamination of several substrates including (R)-α-methylbenzylamine, as well as production of (R)-amines, and determined its crystal structure. The crystal structures of the holoenzyme and the complex with an inhibitor gabaculine offer the first detailed insight into the structural basis for substrate specificity and enantioselectivity of the industrially important class of (R)-selective amine : pyruvate transaminases. Database The atomic coordinates and structure factors for the Nectria TAm in holoenzyme and gabaculine-bound forms have been deposited in the PDB as entries 4cmd and 4cmf respectively. Structured digital abstract • TAm and TAm bind by x-ray crystallography (View interaction) PMID:24618038

  7. 21 CFR 862.1030 - Alanine amino transferase (ALT/SGPT) test system.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Alanine amino transferase (ALT/SGPT) test system... Test Systems § 862.1030 Alanine amino transferase (ALT/SGPT) test system. (a) Identification. An alanine amino transferase (ALT/SGPT) test system is a device intended to measure the activity of the...

  8. 40 CFR 721.520 - Alanine, N-(2-carboxyethyl)-N-alkyl-, salt.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 30 2010-07-01 2010-07-01 false Alanine, N-(2-carboxyethyl)-N-alkyl... Specific Chemical Substances § 721.520 Alanine, N-(2-carboxyethyl)-N-alkyl-, salt. (a) Chemical substance... alanine, N-(2-carboxyethyl)-N- alkyl-, salt (P-89-336) is subject to reporting under this section for the...

  9. 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

  10. 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)

  11. Bioassay-guided fractionation of lemon balm (Melissa officinalis L.) using an in vitro measure of GABA transaminase activity.

    Science.gov (United States)

    Awad, Rosalie; Muhammad, Asim; Durst, Tony; Trudeau, Vance L; Arnason, John T

    2009-08-01

    A novel pharmacological mechanism of action for the anxiolytic botanical Melissa officinalis L. (lemon balm) is reported. The methanol extract was identified as a potent in vitro inhibitor of rat brain GABA transaminase (GABA-T), an enzyme target in the therapy of anxiety, epilepsy and related neurological disorders. Bioassay-guided fractionation led to the identification and isolation of rosmarinic acid (RA) and the triterpenoids, ursolic acid (UA) and oleanolic acid (OA) as active principles. Phytochemical characterization of the crude extract determined RA as the major compound responsible for activity (40% inhibition at 100 microg/mL) since it represented approximately 1.5% of the dry mass of the leaves. Synergistic effects may also play a role. Copyright 2009 John Wiley & Sons, Ltd.

  12. The Alanine Racemase of Mycobacterium smegmatis Is Essential for Growth in the Absence of d-Alanine▿ †

    Science.gov (United States)

    Milligan, Daniel L.; Tran, Sieu L.; Strych, Ulrich; Cook, Gregory M.; Krause, Kurt L.

    2007-01-01

    Alanine racemase, encoded by the gene alr, is an important enzyme in the synthesis of d-alanine for peptidoglycan biosynthesis. Strains of Mycobacterium smegmatis with a deletion mutation of the alr gene were found to require d-alanine for growth in both rich and minimal media. This indicates that alanine racemase is the only source of d-alanine for cell wall biosynthesis in M. smegmatis and confirms alanine racemase as a viable target gene for antimycobacterial drug development. PMID:17827284

  13. Deletion of the Saccharomyces cerevisiae ARO8 gene, encoding an aromatic amino acid transaminase, enhances phenylethanol production from glucose.

    Science.gov (United States)

    Romagnoli, Gabriele; Knijnenburg, Theo A; Liti, Gianni; Louis, Edward J; Pronk, Jack T; Daran, Jean-Marc

    2015-01-01

    Phenylethanol has a characteristic rose-like aroma that makes it a popular ingredient in foods, beverages and cosmetics. Microbial production of phenylethanol currently relies on whole-cell bioconversion of phenylalanine with yeasts that harbour an Ehrlich pathway for phenylalanine catabolism. Complete biosynthesis of phenylethanol from a cheap carbon source, such as glucose, provides an economically attractive alternative for phenylalanine bioconversion. In this study, synthetic genetic array (SGA) screening was applied to identify genes involved in regulation of phenylethanol synthesis in Saccharomyces cerevisiae. The screen focused on transcriptional regulation of ARO10, which encodes the major decarboxylase involved in conversion of phenylpyruvate to phenylethanol. A deletion in ARO8, which encodes an aromatic amino acid transaminase, was found to underlie the transcriptional upregulation of ARO10 during growth, with ammonium sulphate as the sole nitrogen source. Physiological characterization revealed that the aro8Δ mutation led to substantial changes in the absolute and relative intracellular concentrations of amino acids. Moreover, deletion of ARO8 led to de novo production of phenylethanol during growth on a glucose synthetic medium with ammonium as the sole nitrogen source. The aro8Δ mutation also stimulated phenylethanol production when combined with other, previously documented, mutations that deregulate aromatic amino acid biosynthesis in S. cerevisiae. The resulting engineered S. cerevisiae strain produced >3 mm phenylethanol from glucose during growth on a simple synthetic medium. The strong impact of a transaminase deletion on intracellular amino acid concentrations opens new possibilities for yeast-based production of amino acid-derived products. Copyright © 2014 John Wiley & Sons, Ltd.

  14. Proton response of alanine based pellets and films

    Energy Technology Data Exchange (ETDEWEB)

    Onori, S.; De Angelis, C.; Fattibene, P. [Istituto Superiore di Sanita, Rome (Italy). Lab. di Fisica]|[Istituto Nazionale di Fisica Nucleare, Rome (Italy). Sezione Sanita; D`Errico, F. [DCMN, Universita degli Studi di Pisa (Italy); Egger, E. [Paul Scherrer Inst. (PSI), Villigen (Switzerland); Janovsky, I. [Nuclear Research Institute, Rez (Czech Republic)

    1997-01-01

    The performance of two different alanine-ESR systems (1 mm ISS pellets and NRI films) in therapy proton beams was checked against reference dosimetry provided by a Markus parallel plate ionization chamber. Irradiations were performed at the OPTIS facility of the PSI with a 62 MeV proton beam. The energy dependence of the response was derived from the analysis of film stacks irradiated with pristine beams. In fact, 250 {mu}m films allowed for a high resolution sampling of the proton slowing down mechanisms. Moreover pellets were irradiated in unmodulated and modulated beams. Alanine dose distributions were always in good agreement with the PSI Markus chamber results. Simple density scaling allowed for the conversion of the results to the dose distributions in water, consistently with the good tissue equivalence properties of these detectors. (Author).

  15. Degradation of glycine and alanine on irradiated quartz.

    Science.gov (United States)

    Pawlikowski, Maciej; Benko, Aleksandra; Wróbel, Tomasz P

    2013-04-01

    Recent researches suggest participation of minerals in the formation of life under primordial conditions. Among all of the minerals, quartz seems to be one of the most probable to take part in such processes. However, an external source of energy is needed, e.g. electric discharge. A device simulating the proposed conditions was designed and was used to simulate prebiotic conditions. Investigation of processes occurring during the stimulation of quartz with electric discharge was studied by means of Ultraviolet-visible (UV-VIS) spectroscopy, in order to monitor the generation kinetics of free radicals. Additionally, infrared spectroscopy was applied to identify chemical reaction products created in a solution of alanine or glycine, in the presence of quartz treated with electric discharge. Formation of increased amounts of free radicals, compared to experiments performed without quartz and/or amino acid, is reported, along with identification of possible degradation products of alanine. No synthetic reactions were observed.

  16. A thermodynamic study of La(III)L-alanine complexes

    International Nuclear Information System (INIS)

    Elzawawy, F.M.

    1991-01-01

    The protonation constants of L-alanine and the complex formation constants of its La(III) complexes were determined by potentiometric studies at ionic strengths 0.06, 0.1, and 0.15 mol dm -3 (NaClO 4 ) and at different temperatures 20, 27, and 35 O C. The data together with the derived thermodynamic parameters ΔH O , ΔS O , and ΔG O are reported and discussed. (author)

  17. 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

  18. 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

  19. 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

  20. Transport of the alpha-amino-mono-carboxylic acid L-alanine by the beta-alanine carrier of the rabbit ileum

    DEFF Research Database (Denmark)

    Andersen, Vibeke; Munck, B G

    1987-01-01

    The proposal that the beta-alanine carrier of the rabbit ileum is a high affinity carrier of the neutral amino acids was examined by means of measurements of influx across the brush border membrane of the intact epithelium using L-alanine as a representative of the neutral amino acids. Confirming...... the proposal, evidence was provided for mutual competitive inhibition between beta-alanine and L-alanine; and it was also demonstrated that a process contributes to the influx of L-alanine, which is characterized by a maximum rate of transport equal to that of beta-alanine and a Kt, which is equal to the Ki...... of L-alanine against the influx of beta-alanine. In the concentration range 0.01 to 0.125 mM the influx of L-alanine was found to be linearly related to the concentration indicating a significant unstirred layer influence on present and previous estimates of the Kt values for influx of amino acids...

  1. Pancreatic stellate cells support tumour metabolism through autophagic alanine secretion.

    Science.gov (United States)

    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.

  2. Alanine-dependent reactions of 5'-deoxypyridoxal in water.

    Science.gov (United States)

    Go, Maybelle K; Richard, John P

    2008-12-01

    The non-enzymatic reaction of 5'-deoxypyridoxal (DPL) with l-alanine in water at 25 degrees C was investigated. DPL reacts with alanine to form an imine, which then undergoes deprotonation at the alpha-amino carbon of alanine to form a resonance delocalized DPL-stabilized carbanion. At early reaction times the only detectable products are pyruvate and the dimeric species formed by addition of the alpha-pyridine stabilized carbanion to DPL. No Claisen-type products of addition of the alpha-amino carbanion to DPL, as was previously reported to form from the reaction between DPL and glycine [K. Toth, T.L. Amyes, J.P. Richard, J.P.G. Malthouse, M.E. Ni Beilliu, J. Am. Chem. Soc. 126 (2004) 10538-10539], are observed. The electrophile reacts instead at the alpha-pyridyl carbon. This dimer is in chemical equilibrium with reactants. At longer reaction times about 50% of DPL is converted to 5'-deoxypyridoxamine, the thermodynamically favored product of formal transamination of DPL.

  3. 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.

  4. 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)

  5. Phospholipid transfer protein activity is determined by type 2 diabetes mellitus and metabolic syndrome, and is positively associated with serum transaminases

    NARCIS (Netherlands)

    Dullaart, Robin P. F.; de Vries, Rindert; Dallinga-Thie, Geesje M.; Sluiter, Wim J.; van Tol, Arie

    2008-01-01

    BACKGROUND: The extent to which plasma phospholipid transfer protein (PLTP) activity is affected by type 2 diabetes mellitus (DM) and metabolic syndrome (MetS) is still unknown. PLTP is synthesized in the liver, and elevated serum transaminases are considered to predict nonalcoholic fatty liver

  6. Phospholipid transfer protein activity is determined by type 2 diabetes mellitus and metabolic syndrome, and is positively associated with serum transaminases

    NARCIS (Netherlands)

    Dullaart, Robin P. F.; de Vries, Rindert; Dallinga-Thie, Geesje M.; Sluiter, Wim J.; van Tol, Arie

    Background The extent to which plasma phospholipid transfer protein (PLTP) activity is affected by type 2 diabetes mellitus (DM) and metabolic syndrome (MetS) is still unknown. PLTP is synthesized in the liver, and elevated serum transaminases are considered to predict nonalcoholic fatty liver

  7. SIGNIFICANCE OF LACTATE DEHYDROGENASE AND ASPARTATE TRANSAMINASE AS BIOCHEMICAL MARKERS AND AS PREDICTORS OF SEVERITY OF PREGNANCY-INDUCED HYPERTENSION AND ITS COMPLICATIONS

    Directory of Open Access Journals (Sweden)

    Ramesh Sonowal

    2017-03-01

    Full Text Available BACKGROUND To compare serum Lactate Dehydrogenase (LDH and serum Aspartate Transaminase (AST of normotensive pregnant women with those of preeclamptic and eclamptic women. To determine the relationship of levels of serum lactate dehydrogenase and serum aspartate transaminase with severity of pregnancy-induced hypertension and its complications. MATERIALSAND METHODS The study was carried out on pregnant hypertensive patients attending outpatient department of Obstetrics and Gynaecology department, AMCH, Dibrugarh, Assam from 1 st July 2013 to 30 th June 2014. Normotensive pregnant women were taken as controls. Each serum sample from both the control group as well as study group was estimated for lactate dehydrogenase and aspartate transaminase using standard methods and a comparison is drawn and analysed using t-test and Chi-square test. RESULTS Serum lactate dehydrogenase and serum aspartate transaminase levels were higher in the study group in comparison to the study groups. The mean serum LDH was 198±30.03U/L in control group, whereas in preeclampsia and eclampsia, mean serum levels of LDH were 817±114U/L and 927±108U/L, respectively. The levels of the serum AST were found to be less than 600U/L in normotensive and preeclampsia patients and more than 600 U/L in eclampsia and other complications of PIH. CONCLUSION Serum lactate dehydrogenase and serum aspartate transaminase levels in patients suffering from preeclampsia and its complications are consistently higher compared to the normotensive pregnant patients. To determine the usefulness of inclusion of these enzymes along with other cardiac enzymes in the panel of investigations of pregnant women universally needs further large scale comparative studies.

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

    Directory of Open Access Journals (Sweden)

    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

  9. 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

  10. The mechanisms of radical formation in L-α-alanine

    International Nuclear Information System (INIS)

    Bugay, A.A.; Onischuk, V.A.; Petrenko, T.L.; Teslenko, V.V.

    2000-01-01

    Modeling of radical transformations in L-α-alanine after irradiation was performed for isolated radicals and for clusters. Special attention was devoted to the explanation of the experimental results concerning selective proton transfer and behavior of cation-radicals because a unique interpretation of the corresponding experiments is very difficult. Both semi-empirical and ab initio methods were used depending on the size of system under investigation. The results obtained show the usefulness of the computer simulation for processes in rather complex materials used in dosimetry

  11. Prevalence, knowledge and attitudes relating to β-alanine use among professional footballers.

    Science.gov (United States)

    Kelly, Vincent G; Leveritt, Michael D; Brennan, Christopher T; Slater, Gary J; Jenkins, David G

    2017-01-01

    To investigate β-alanine supplementation use and level of knowledge amongst professional footballers. Cross-sectional survey of Australian professional football players. Questionnaires assessing β-alanine supplementation behaviours, level of knowledge and sources of information were completed by professional rugby union (RU) (n=87), rugby league (RL) (n=180) and Australian Rules Football (ARF) (n=303) players. Approximately 61% of athletes reported β-alanine use, however use by ARF football players (44%) was lower than that of RU (80%) and RL players (80%). The majority of respondents were not using β-alanine in accordance with recommendations. Only 35% of the participants were able to correctly identify the potential benefits of β-alanine supplementation. The main information sources that influenced players' decision to use β-alanine were strength and conditioning coach (71%) and dietitian (52%). Forty-eight per cent of athletes never read labels prior to supplementing and only 11% completed their own research on β-alanine. Compared to RL and ARF players, RU players had both a greater knowledge of β-alanine supplementation and better supplementation practices. Despite over half the surveyed professional footballers using β-alanine, the majority of athletes used β-alanine in a manner inconsistent with recommendations. A better understanding of the environment and culture within professional football codes is required before supplement use becomes consistent with evidence based supplement recommendations. Copyright © 2016 Sports Medicine Australia. Published by Elsevier Ltd. All rights reserved.

  12. An Agrobacterium tumefaciens Strain with Gamma-Aminobutyric Acid Transaminase Activity Shows an Enhanced Genetic Transformation Ability in Plants

    Science.gov (United States)

    Nonaka, Satoko; Someya, Tatsuhiko; Zhou, Sha; Takayama, Mariko; Nakamura, Kouji; Ezura, Hiroshi

    2017-01-01

    Agrobacterium tumefaciens has the unique ability to mediate inter-kingdom DNA transfer, and for this reason, it has been utilized for plant genetic engineering. To increase the transformation frequency in plant genetic engineering, we focused on gamma-aminobutyric acid (GABA), which is a negative factor in the Agrobacterium-plant interaction. Recent studies have shown contradictory results regarding the effects of GABA on vir gene expression, leading to the speculation that GABA inhibits T-DNA transfer. In this study, we examined the effect of GABA on T-DNA transfer using a tomato line with a low GABA content. Compared with the control, the T-DNA transfer frequency was increased in the low-GABA tomato line, indicating that GABA inhibits T-DNA transfer. Therefore, we bred a new A. tumefaciens strain with GABA transaminase activity and the ability to degrade GABA. The A. tumefaciens strain exhibited increased T-DNA transfer in two tomato cultivars and Erianthus arundinacues and an increased frequency of stable transformation in tomato. PMID:28220841

  13. Plasma membrane fatty acid-binding protein and mitochondrial glutamic-oxaloacetic transaminase of rat liver are related

    Energy Technology Data Exchange (ETDEWEB)

    Berk, P.D.; Potter, B.J.; Sorrentino, D.; Zhou, S.L.; Isola, L.M.; Stump, D.; Kiang, C.L.; Thung, S. (Mount Sinai School of Medicine, New York, NY (USA)); Wada, H.; Horio, Y. (Univ. of Osaka (Japan))

    1990-05-01

    The hepatic plasma membrane fatty acid-binding protein (h-FABP{sub PM}) and the mitochondrial isoenzyme of glutamic-oxaloacetic transaminase (mGOT) of rat liver have similar amino acid compositions and identical amino acid sequences for residues 3-24. Both proteins migrate with an apparent molecular mass of 43 kDa on SDS/polyacrylamide gel electrophoresis, have a similar pattern of basic charge isomers on isoelectric focusing, are eluted similarly from four different high-performance liquid chromatographic columns, have absorption maxima at 435 nm under acid conditions and 354 nm at pH 8.3, and bind oleate. Sinusoidally enriched liver plasma membranes and purified h-FABP{sub PM} have GOT enzymatic activity. Monospecific rabbit antiserum against h-FABP{sub PM} reacts on Western blotting with mGOT, and vice versa. Antisera against both proteins produce plasma membrane immunofluorescence in rat hepatocytes and selectively inhibit the hepatocellular uptake of ({sup 3}H)oleate but not that of ({sup 35}S)sulfobromophthalein or ({sup 14}C)taurocholate. The inhibition of oleate uptake produced by anti-h-FABP{sub PM} can be eliminated by preincubation of the antiserum with mGOT; similarly, the plasma membrane immunofluorescence produced by either antiserum can be eliminated by preincubation with the other antigen. These data suggest that h-FABP{sub PM} and mGOT are closely related.

  14. An Agrobacterium tumefaciens Strain with Gamma-Aminobutyric Acid Transaminase Activity Shows an Enhanced Genetic Transformation Ability in Plants.

    Science.gov (United States)

    Nonaka, Satoko; Someya, Tatsuhiko; Zhou, Sha; Takayama, Mariko; Nakamura, Kouji; Ezura, Hiroshi

    2017-02-21

    Agrobacterium tumefaciens has the unique ability to mediate inter-kingdom DNA transfer, and for this reason, it has been utilized for plant genetic engineering. To increase the transformation frequency in plant genetic engineering, we focused on gamma-aminobutyric acid (GABA), which is a negative factor in the Agrobacterium-plant interaction. Recent studies have shown contradictory results regarding the effects of GABA on vir gene expression, leading to the speculation that GABA inhibits T-DNA transfer. In this study, we examined the effect of GABA on T-DNA transfer using a tomato line with a low GABA content. Compared with the control, the T-DNA transfer frequency was increased in the low-GABA tomato line, indicating that GABA inhibits T-DNA transfer. Therefore, we bred a new A. tumefaciens strain with GABA transaminase activity and the ability to degrade GABA. The A. tumefaciens strain exhibited increased T-DNA transfer in two tomato cultivars and Erianthus arundinacues and an increased frequency of stable transformation in tomato.

  15. Electron beam dose measurements with alanine/ESR dosimeter

    International Nuclear Information System (INIS)

    Rodrigues, O. Jr.; Galante, O.L.; Campos, L.L.

    2001-01-01

    When the aminoacid alanine, CH 3 -CH(NH 2 )-COOH, is exposed to radiation field, stable free radicals are produced. The predominant paramagnetic specie found at room temperature is the CH 3 -CH-COOH. Electron Spin Resonance - ESR is a technique used for quantification and analysis of radicals in solid and liquid samples. The evaluation of the amount of produced radicals can be associated with the absorbed dose . The alanine/ESR is an established dosimetry method employed for high doses evaluation, it presents good performance for X-rays, gamma, electrons, and protons radiation detection. The High Doses Dosimetry Laboratory of Ipen developed a dosimetric system based on alanina/ESR that presents good characteristics for use in gamma fields such as: wide dose range from 10 to 10 5 Gy, low fading, low uncertainty (<5%), no dose rate dependence and non-destructive ESR single readout. The detector is encapsulated in a special polyethylene tube that reduces the humidity problems and improves the mechanical resistance. The IPEN dosimeter was investigated for application in electron beam fields dosimetry

  16. Dosimetry of blood irradiation using an alanine/ESR dosimeter

    International Nuclear Information System (INIS)

    Chen, F.; Covas, D.T.; Baffa, O.

    2001-01-01

    A batch of 80 DL-alanine dosimeters was supplied to Hemocentro of the Hospital and Clinics of Faculdade de Medicina de Ribeirao Preto (HC-FMRP) SP, Brazil for the purpose of quality control of the radiation dose delivered to blood bags. The irradiation was made using two (40x40) cm 2 parallel opposed radiation fields each with 80 cm of source to surface distance in the Radiotherapy Section of HC-FMRP with the 60 Co teletherapy unit. The calculated radiation absorbed dose at the center of the box was 20 Gy. The dosimeter readings were performed using a Varian E-4 ESR Spectrometer operating in X-band. For the 80 dosimeters and over the irradiation volume throughout a blood bag, the minimum and maximum doses were 14 and 23 Gy, respectively. The mean dose was (18±2) Gy (1σ), and the coefficient of variability was 11.1%. Alanine dosimeters demonstrated easy handling, good precision and adequate sensitivity for this application

  17. The effect of immunonutrition (glutamine, alanine on fracture healing

    Directory of Open Access Journals (Sweden)

    Abdullah Küçükalp

    2014-11-01

    Full Text Available Background: There have been various studies related to fracture healing. Glutamine is an amino acid with an important role in many cell and organ functions. This study aimed to make a clinical, radiological, and histopathological evaluation of the effects of glutamine on fracture healing. Methods: Twenty rabbits were randomly allocated into two groups of control and immunonutrition. A fracture of the fibula was made to the right hind leg. All rabbits received standard food and water. From post-operative first day for 30 days, the study group received an additional 2 ml/kg/day 20% L-alanine L-glutamine solution via a gastric catheter, and the control group received 2 ml/kg/day isotonic via gastric catheter. At the end of 30 days, the rabbits were sacrificed and the fractures were examined clinically, radiologically, and histopathologically in respect to the degree of union. Results: Radiological evaluation of the control group determined a mean score of 2.5 according to the orthopaedists and 2.65 according to the radiologists. In the clinical evaluation, the mean score was 1.875 for the control group and 2.0 for the study group. Histopathological evaluation determined a mean score of 8.5 for the control group and 9.0 for the study group. Conclusion: One month after orally administered glutamine–alanine, positive effects were observed on fracture healing radiologically, clinically, and histopathologically, although no statistically significant difference was determined.

  18. Recurrent truncating mutations in alanine-glyoxylate aminotransferase gene in two South Indian families with primary hyperoxaluria type 1 causing later onset end-stage kidney disease

    Directory of Open Access Journals (Sweden)

    A K Dutta

    2016-01-01

    Full Text Available Primary hyperoxaluria type 1 is an autosomal recessive inborn error of metabolism due to liver-specific peroxisomal enzyme alanine-glyoxylate transaminase deficiency. Here, we describe two unrelated patients who were diagnosed to have primary hyperoxaluria. Homozygous c.445_452delGTGCTGCT (p.L151Nfs*14 (Transcript ID: ENST00000307503; human genome assembly GRCh38.p2 (HGMD ID CD073567 mutation was detected in both the patients and the parents were found to be heterozygous carriers. Our patients developed end-stage renal disease at 23 years and 35 years of age. However, in the largest series published from OxalEurope cohort, the median age of end-stage renal disease for null mutations carriers was 9.9 years, which is much earlier than our cases. Our patients had slower progressions as compared to three unrelated patients from North India and Pakistan, who had homozygous c.302T>C (p.L101P (HGMD ID CM093792 mutation in exon 2. Further, patients need to be studied to find out if c.445_452delGTGCTGCT mutation represents a founder mutation in Southern India.

  19. Eukaryotic beta-alanine synthases are functionally related but have a high degree of structural diversity

    DEFF Research Database (Denmark)

    Gojkovic, Zoran; Sandrini, Michael; Piskur, Jure

    2001-01-01

    beta -Alanine synthase (EC 3.5.1.6), which catalyzes the final step of pyrimidine catabolism, has only been characterized in mammals. A Saccharomyces kluyveri pyd3 mutant that is unable to grow on N-carbamy-beta -alanine as the sole nitrogen source and exhibits diminished beta -alanine synthase...... no pyrimidine catabolic pathway, it enabled growth on N-carbamyl- beta -alanine as the sole nitrogen source. The D. discoideum and D. melanogaster PYD3 gene products are similar to mammalian beta -alanine synthases. In contrast, the S. kluyveri protein is quite different from these and more similar to bacterial...... N- carbamyl amidohydrolases. All three beta -alanine synthases are to some degree related to various aspartate transcarbamylases, which catalyze the second step of the de novo pyrimidine biosynthetic pathway. PYD3 expression in yeast seems to be inducible by dihydrouracil and N...

  20. Structural features and kinetic characterization of alanine racemase from Bacillus pseudofirmus OF4.

    Science.gov (United States)

    Dong, Hui; Hu, Tingting; He, Guangzheng; Lu, Deren; Qi, Jianxun; Dou, Yanshu; Long, Wei; He, Xin; Ju, Jiansong; Su, Dan

    2018-02-26

    Alanine racemase (Alr) is a pyridoxal-5'-phosphate-dependent (PLP) enzyme that catalyzes a reversible racemization between the enantiomers of alanine. d-Alanine is an indispensable constituent in the biosynthesis of bacterial cell-wall peptidoglycan, and its inhibition is lethal to prokaryotes, which makes it an attractive target for designing antibacterial drugs. In this study, the molecular structure of alanine racemase from Bacillus pseudofirmus OF4 (DadX OF4 ) was determined by X-ray crystallography to a resolution of 1.8 Å. The comparison of DadX OF4 with alanine racemases from other bacteria demonstrated a conserved overall fold. Enzyme kinetics analysis showed that the conserved residues at the substrate entryway and the salt bridge at the dimer interface are critical for enzyme activity. These structural and biochemical findings provide a template for future structure-based drug-development efforts targeting alanine racemases. Copyright © 2018 Elsevier Inc. All rights reserved.

  1. Involvement of alanine racemase in germination of Bacillus cereus spores lacking an intact exosporium.

    Science.gov (United States)

    Venir, Elena; Del Torre, Manuela; Cunsolo, Vincenzo; Saletti, Rosaria; Musetti, Rita; Stecchini, Mara Lucia

    2014-02-01

    The L-alanine mediated germination of food isolated Bacillus cereus DSA 1 spores, which lacked an intact exosporium, increased in the presence of D-cycloserine (DCS), which is an alanine racemase (Alr) inhibitor, reflecting the activity of the Alr enzyme, capable of converting L-alanine to the germination inhibitor D-alanine. Proteomic analysis of the alkaline extracts of the spore proteins, which include exosporium and coat proteins, confirmed that Alr was present in the B. cereus DSA 1 spores and matched to that encoded by B. cereus ATCC 14579, whose spore germination was strongly affected by the block of conversion of L- to D-alanine. Unlike ATCC 14579 spores, L-alanine germination of B. cereus DSA 1 spores was not affected by the preincubation with DCS, suggesting a lack of restriction in the reactant accessibility.

  2. Application of an alanine dosimetry system for industrial irradiation and radiation protection

    International Nuclear Information System (INIS)

    Gohs, U.

    1996-01-01

    This paper reports the application of alanine dosimetry in radiation processing. Continuous checks of the EPR measuring conditions as well as using high-quality alanine dosimeters and consistent technique for dose determination guarantee an accuracy of about ± 3% intermediate dose levels. The alanine dosimetry system was applied for dose mapping measurements during irradiator qualification and performance qualification of different products, routine dosimetry, and special radiation protection applications within the gamma irradiator. (author)

  3. 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.)

  4. Alanine Enhances Aminoglycosides-Induced ROS Production as Revealed by Proteomic Analysis

    Directory of Open Access Journals (Sweden)

    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.

  5. 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)

  6. Mutants of GABA transaminase (POP2 suppress the severe phenotype of succinic semialdehyde dehydrogenase (ssadh mutants in Arabidopsis.

    Directory of Open Access Journals (Sweden)

    Frank Ludewig

    Full Text Available BACKGROUND: The gamma-aminubutyrate (GABA shunt bypasses two steps of the tricarboxylic acid cycle, and is present in both prokaryotes and eukaryotes. In plants, the pathway is composed of the calcium/calmodulin-regulated cytosolic enzyme glutamate decarboxylase (GAD, the mitochondrial enzymes GABA transaminase (GABA-T; POP2 and succinic semialdehyde dehydrogenase (SSADH. We have previously shown that compromising the function of the GABA-shunt, by disrupting the SSADH gene of Arabidopsis, causes enhanced accumulation of reactive oxygen intermediates (ROIs and cell death in response to light and heat stress. However, to date, genetic investigations of the relationships between enzymes of the GABA shunt have not been reported. PRINCIPAL FINDINGS: To elucidate the role of succinic semialdehyde (SSA, gamma-hydroxybutyrate (GHB and GABA in the accumulation of ROIs, we combined two genetic approaches to suppress the severe phenotype of ssadh mutants. Analysis of double pop2 ssadh mutants revealed that pop2 is epistatic to ssadh. Moreover, we isolated EMS-generated mutants suppressing the phenotype of ssadh revealing two new pop2 alleles. By measuring thermoluminescence at high temperature, the peroxide contents of ssadh and pop2 mutants were evaluated, showing that only ssadh plants accumulate peroxides. In addition, pop2 ssadh seedlings are more sensitive to exogenous SSA or GHB relative to wild type, because GHB and/or SSA accumulate in these plants. SIGNIFICANCE: We conclude that the lack of supply of succinate and NADH to the TCA cycle is not responsible for the oxidative stress and growth retardations of ssadh mutants. Rather, we suggest that the accumulation of SSA, GHB, or both, produced downstream of the GABA-T transamination step, is toxic to the plants, resulting in high ROI levels and impaired development.

  7. Valores de transaminasas en cabras criollas infectadas con Trypanosoma vivax Transaminases values in Creole goats infected with Trypanosoma vivax

    Directory of Open Access Journals (Sweden)

    Emir Espinoza

    2002-01-01

    Full Text Available La presente comunicación reporta los valores de las enzimas transaminasas, Aspartatoaminotransferasa (AST y Alaninaaminotransferasa (ALT encontrados en sueros de cabras infectadas con la cepa de Trypanosoma vivax Stock (TvIIV y sus controles. Las determinaciones se realizaron durante un lapso experimental de diez semanas, divididos en dos períodos iguales (pre y post-infección por intermedio de un método colorimétrico, utilizando kits comerciales. Los datos fueron analizados mediante la prueba t Student's. En el caso de la AST, la comparación de las medias parciales de ambos grupos infectado y control, no indicó diferencias estadísticas. Con respecto a la ALT, la contrastación de las medias parciales de pre y post-infección del grupo de cabras infectadas, señaló diferencias significativas (PThe present communication reports the transaminases enzymes values Aspartatoaminotransferase (AST and Alaninaaminotransferase (ALT in serum from goats infected with the Trypanosoma vivax Stock (TvIIV. The determinations were realized during a ten week experimental period divided into two equal periods (pre- and post-infection by colorimetric method, using commercial kits. The dates were analyzed through the t Student's test. In the AST case, the comparison between partial means of infected and control groups did not show any statistical differences. In relation to ALT, the contrast of partial means to pre- and post-infection from infected goats group indicated significant differences (P<0.01.

  8. Resistance imparted by traditional Chinese medicines to the acute change of glutamic pyruvic transaminase, alkaline phosphatase and creatine kinase activities in rat blood caused by noise.

    Science.gov (United States)

    Zhu, Bei-Wei; Sun, Yu-Mei; Yun, Xia; Han, Song; Piao, Mei-Lan; Murata, Yoshiyuki; Tada, Mikiro

    2004-05-01

    The activities of serum glutamic pyruvic transaminase (GPT), alkaline phosphatase (ALP) and creatine kinase (CK) in rats injected or not with the Chinese medicines, Astragali, Rhodiolae and Ligusticum, were determined after noise exposure. Noise at 95 and 105 dB significantly increased the activities of GPT, ALP and CK, and showed a dependence on the exposure time. The injection of each medicine significantly suppressed the increased enzyme activities by 95 and 105 dB noise.

  9. The Response of Alanine Dosimeters in Thermal Neutron Fields

    DEFF Research Database (Denmark)

    Schmitz, T.; Bassler, Niels; Sharpe, P.

    Purpose: Boron Neutron Capture Therapy (BNCT) is a special kind of particle therapy, based on the neutron induced fission of the boron isotope 10B [1]. We have performed dosimetry experiments on the mixed neutron and gamma fields at the TRIGA Mark II research reactor in Mainz. Commonly, dosimetry...... in such fields is realized by foil activation and ion chambers [2]. Here we investigate alanine as an easier and more robust alternative dosimeter. Methods: We have performed four phantom experiments at the TRIGA Mark II research reactor in Mainz [3], in a predominantly thermal neutron field with a strong gamma...... response of all pellets could be reproduced by calculations within a uncertainty of 5 %. For all experiments three dose components have been separated. A proton dose is generated in the 14N(n,p)14C reaction. Secondary gammas are generated by various (n,γ) reactions, dominated by the 2.2 MeV photon from...

  10. Elaboration And Study Of Transfer Alanine/ESR Dosemeters

    International Nuclear Information System (INIS)

    Torres G, Luz A.

    1996-01-01

    The dosimetry is the dose measure imparted by the energy from the ionizing radiation to the matter. The dosemeters is the means used for the determination of such a dose. Diverse dosimetric classes exist, this classification depends in essence of the energy involved in the irradiation process and of its application necessity. It is as well as in radiological protection movie dosemeter is used, and TLD, in the calibration of units, as those of cobalt, the ionization cameras are used, in the detection of superficial contamination the accountants Geiger Muller and proportional etc. The transfer dosimeter Alanine/ESR is used, object of the present work, is characterized because after reading the registered dose, the information is conserved and it can be transferred and read in any team of ESR. Likewise the following dose that is imparted will be added to the previous one registered, this indicates that the dosemeters is of integral character. In the spectra taken ESR, it is determined that the integral double or area under the curve of the spectrum ESR is proportional to the concentration of free radicals generated by the radiation ionization and in turn this concentration is proportional to the dose received by the pills; in last, these proportionality relationships take to that the area under the curve of the spectrum main ESR is proportional to the radiation dose received by the alanine dosemeters. This dosemeter seeks to be an economic alternative that it will lend the calibration service that today offers the ionization Cameras or the acrylic red industrial dosemeters

  11. 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.

  12. A comparison of three materials used in ESR dosimetry: L-α-alanine, DL-α-alanine and standard bone powder. Response to Co-60 gamma radiation

    International Nuclear Information System (INIS)

    Stuglik, Z.; Sadlo, J.

    1995-01-01

    Three solid state materials: L-α-alanine, DL-α-alanine and standard bone powder were irradiated with gamma analyzed with ESR method. It was stated that the G-value of paramagnetic centres in L-α-alanine is practically the same as in DL-alpha-alanine and about 50 times higher than in non-deproteinized bone powder. The sensitivities of investigated materials are proportional to their G-values if double integrals of ESR signals are chosen as a measure of radiation effects. When first derivatives of ESR absorption bands are used to the construction of dose-response curves (peak-to-peak method) the sensitivities of all investigated materials are comparable. (author). 14 refs, 1 fig., 3 tabs

  13. 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.

  14. 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.

  15. Regulation of the ald Gene Encoding Alanine Dehydrogenase by AldR in Mycobacterium smegmatis

    Science.gov (United States)

    Jeong, Ji-A; Baek, Eun-Young; Kim, Si Wouk; Choi, Jong-Soon

    2013-01-01

    The regulatory gene aldR was identified 95 bp upstream of the ald gene encoding l-alanine dehydrogenase in Mycobacterium smegmatis. The AldR protein shows sequence similarity to the regulatory proteins of the Lrp/AsnC family. Using an aldR deletion mutant, we demonstrated that AldR serves as both activator and repressor for the regulation of ald gene expression, depending on the presence or absence of l-alanine. The purified AldR protein exists as a homodimer in the absence of l-alanine, while it adopts the quaternary structure of a homohexamer in the presence of l-alanine. The binding affinity of AldR for the ald control region was shown to be increased significantly by l-alanine. Two AldR binding sites (O1 and O2) with the consensus sequence GA-N2-ATC-N2-TC and one putative AldR binding site with the sequence GA-N2-GTT-N2-TC were identified upstream of the ald gene. Alanine and cysteine were demonstrated to be the effector molecules directly involved in the induction of ald expression. The cellular level of l-alanine was shown to be increased in M. smegmatis cells grown under hypoxic conditions, and the hypoxic induction of ald expression appears to be mediated by AldR, which senses the intracellular level of alanine. PMID:23749971

  16. Polymerization of alanine in the presence of a non-swelling montmorillonite

    Science.gov (United States)

    Paecht-Horowitz, M.; Lahav, N.

    1977-01-01

    Alanine, starting from alanine-adenylate, has been polymerized in the presence of non-swelling Al-montmorillonite. The yield of polymerization is much lower than that obtained in the presence of swelling Na-montmorillonite. The possibility that the changing interlayer spacing in Na-montmorillonite might be responsible for its catalytic properties, is discussed.

  17. Alanine dosimetry at NPL - the development of a mailed reference dosimetry service at radiotherapy dose levels

    International Nuclear Information System (INIS)

    Sharpe, P.H.G.; Sephton, J.P.

    1999-01-01

    In this paper we describe the work that has been carried out at National Physical Laboratory (NPL) to develop a mailed alanine reference dosimetry service for radiotherapy dose levels. The service is based on alanine/paraffin wax dosimeters produced at NPL. Using a data analysis technique based on spectrum fitting, it has been possible to achieve a precision of dose measurement better than ±0.05 Gy (1σ). A phantom set has been developed for use in high energy photon beams, which enables simultaneous irradiation of alanine dosimeters and ionisation chambers in a well defined geometry. Studies in photon beams of energies between 60 Co and 20 MeV have shown no significant energy dependence (<1%) for alanine relative to dose determination using a graphite calorimeter. Work is underway to extend the service to electron beams, and preliminary results are presented on the direct calibration of alanine in electron beams using a graphite calorimeter. (author)

  18. Synthesis and Analysis of Methacryloyl-L-Alanine Methyl Ester using fourier Transform Nuclear Magnetic Resonance

    International Nuclear Information System (INIS)

    Tri Darwinto

    2008-01-01

    Methacryloyl-L-alanine methyl ester was synthesized by reacting methacrylic acid with L-alanine methyl ester hydrochloride in triethylamine at temperature of 90 o C. Hydrogel polymer of poly(methacryloyl-L-alanine methyl ester) was much used for diagnosis and therapy of vascular tumor. The molecular structure methacryloyl-L-alanine methyl ester analyzed by fourier transform nuclear magnetic resonance (FT-NMR) for analyzing of carbon atom ( 13 C) using Distortionless Enhancement by Polarization Transfer (DEPT) measurement mode with coupling as well as without coupling from proton atom ( 1 H). Molecular structure analysis result showed that DEPT FT-NMR measurement mode with coupling as well as without coupling from 1 H was very fast, exact and accurate method for molecular analysis of organic compound especially methacryloyl-L-alanine methyl ester. (author)

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

    Science.gov (United States)

    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.

  20. 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)

  1. Brain alanine formation as an ammonia-scavenging pathway during hyperammonemia

    DEFF Research Database (Denmark)

    Dadsetan, Sherry; Kukolj, Eva; Bak, Lasse Kristoffer

    2013-01-01

    and cerebral edema. However, ammonia may also be detoxified by the concerted action of glutamate dehydrogenase (GDH) and alanine aminotransferase (ALAT) leading to trapping of ammonia in alanine, which in vivo likely leaves the brain. Our aim was to investigate whether the GS inhibitor methionine sulfoximine...... (MSO) enhances incorporation of (15)NH4(+) in alanine during acute hyperammonemia. We observed a fourfold increased amount of (15)NH4 incorporation in brain alanine in rats treated with MSO. Furthermore, co-cultures of neurons and astrocytes exposed to (15)NH4Cl in the absence or presence of MSO...... demonstrated a dose-dependent incorporation of (15)NH4 into alanine together with increased (15)N incorporation in glutamate. These findings provide evidence that ammonia is detoxified by the concerted action of GDH and ALAT both in vivo and in vitro, a mechanism that is accelerated in the presence of MSO...

  2. Preliminary assessment of LiF and alanine detectors for the dosimetry of proton therapy beams

    International Nuclear Information System (INIS)

    Fattibene, P.; Calicchia, A.; De Angelis, C.; Onori, S.; Egger, E.

    1996-01-01

    An experimental intercomparison between the proton response of LiF TLD-100 and alanine detectors is reported. The investigations were performed with LiF chips and alanine pellets in a 62 MeV proton beam at the Paul Scherrer Institut in Villigen (CH). Results were compared with reference dosimetry provided by Markus type parallel plate ionization chamber. The response of the detectors was studied, in a phantom, at different beam penetration depths in pristine and modulated beams. For both alanine and TL detectors, within the experimental uncertainty of the measurements, no significant energy dependence in the response was observed down to the Bragg peak region. The sensitivity of alanine and LiF detectors to protons was measured in the centre of modulated Bragg peak and no significant difference was found with respect to 60 Co. Contrary to LiF, alanine also offers a remarkable tissue equivalence which favours its choice for in-phantom dosimetry. (author)

  3. 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

  4. Uses of polymer-alanine film/ESR dosimeters in dosimetry of ionizing radiation

    International Nuclear Information System (INIS)

    Xie Liqing; Zhang Yinfeng; Dai Jinxian; Lu Ting; Chen Ruyi; Yang Hua

    1993-01-01

    Alanine ESR dosimetry is a reliable method, used in a various fields of ionizing radiation. The polymer-alanine film/ESR dosimeters of 0.3 -0.4 mm thickness were prepared and their dosimetric properties were studied for 60 Co γ photons and 3 - 5 MeV electrons in the dose range from 20 Gy to 100 kGy. The results show that under normal conditions the alanine calibration curves are linear in the dose range from 100 Gy to 10kGy. The dose profiles at the electron radiation field were measured with the film alanine dosimeters. The polymer-alanine film dosimeters were used for ion implantation of 400 keV ion implantor. Their dose response and energy dependence were investigated initially. (Author)

  5. Thermal decomposition behavior of potassium and sodium jarosite synthesized in the presence of methylamine and alanine

    Energy Technology Data Exchange (ETDEWEB)

    J. Michelle Kotler; Nancy W. Hinman; C. Doc Richardson; Jill R. Scott

    2010-10-01

    Biomolecules, methylamine and alanine, found associated with natural jarosite samples peaked the interest of astrobiologists and planetary geologists. How the biomolecules are associated with jarosite remains unclear although the mechanism could be important for detecting biosignatures in the rock record on Earth and other planets. A series of thermal gravimetric experiments using synthetic K-jarosite and Na-jarosite were conducted to determine if thermal analysis could differentiate physical mixtures of alanine and methylamine with jarosite from samples where the methylamine or alanine was incorporated into the synthesis procedure. Physical mixtures and synthetic experiments with methylamine and alanine could be differentiated from one another and from the standards by thermal analysis for both the K-jarosite and Na-jarosite end-member suites. Changes included shifts in on-set temperatures, total temperature changes from on-set to final, and the presence of indicator peaks for methylamine and alanine in the physical mixture experiments.

  6. Impact of charged amino acid substitution in the transmembrane domain of L-alanine exporter, AlaE, of Escherichia coli on the L-alanine export.

    Science.gov (United States)

    Kim, Seryoung; Ihara, Kohei; Katsube, Satoshi; Ando, Tasuke; Isogai, Emiko; Yoneyama, Hiroshi

    2017-01-01

    The Escherichia coli alaE gene encodes the L-alanine exporter, AlaE, that catalyzes active export of L-alanine using proton electrochemical potential. The transporter comprises only 149 amino acid residues and four predicted transmembrane domains (TMs), which contain three charged amino acid residues. The AlaE-deficient L-alanine non-metabolizing cells (ΔalaE cells) appeared hypersusceptible to L-alanyl-L-alanine showing a minimum inhibitory concentration (MIC) of 2.5 µg/ml for the dipeptide due to a toxic accumulation of L-alanine. To elucidate the mechanism by which AlaE exports L-alanine, we replaced charged amino acid residues in the TMs, glutamic acid-30 (TM-I), arginine-45 (TM-II), and aspartic acid-84 (TM-III) with their respective charge-conserved amino acid or a net neutral cysteine. The ΔalaE cells producing R45K or R45C appeared hypersusceptible to the dipeptide, indicating that arginine-45 is essential for AlaE activity. MIC of the dipeptide in the ΔalaE cells expressing E30D and E30C was 156 µg/ml and >10,000 µg/ml, respectively, thereby suggesting that a negative charge at this position is not essential. The ΔalaE cells expressing D84E or D84C showed an MIC >10,000 and 78 µg/ml, respectively, implying that a negative charge is required at this position. These results were generally consistent with that of the L-alanine accumulation experiments in intact cells. We therefore concluded that charged amino acid residues (R45 and D84) in the AlaE transmembrane domain play a pivotal role in L-alanine export. Replacement of three cysteine residues at C22, C28 (both in TM-I), and C135 (C-terminal region) with alanine showed only a marginal effect on L-alanine export.

  7. Effect of beta-alanine, with and without sodium bicarbonate, on 2000-m rowing performance.

    Science.gov (United States)

    Hobson, Ruth M; Harris, Roger C; Martin, Dan; Smith, Perry; Macklin, Ben; Gualano, Bruno; Sale, Craig

    2013-10-01

    To examine the effect of beta-alanine only and beta-alanine with sodium bicarbonate supplementation on 2,000-m rowing performance. Twenty well-trained rowers (age 23 ± 4 y; height 1.85 ± 0.08 m; body mass 82.5 ± 8.9 kg) were assigned to either a placebo or beta-alanine (6.4 g · d(-1) for 4 weeks) group. A 2,000-m rowing time trial (TT) was performed before supplementation (Baseline) and after 28 and 30 days of supplementation. The post supplementation trials involved supplementation with either maltodextrin or sodium bicarbonate in a double-blind, crossover design, creating four study conditions (placebo with maltodextrin; placebo with sodium bicarbonate; beta-alanine with maltodextrin; beta-alanine with sodium bicarbonate). Blood lactate, pH, bicarbonate, and base excess were measured pre-TT, immediately post-TT and at TT+5 min. Performance data were analyzed using magnitude based inferences. Beta-alanine supplementation was very likely to be beneficial to 2,000-m rowing performance (6.4 ± 8.1 s effect compared with placebo), with the effect of sodium bicarbonate having a likely benefit (3.2 ± 8.8 s). There was a small (1.1 ± 5.6 s) but possibly beneficial additional effect when combining chronic beta-alanine supplementation with acute sodium bicarbonate supplementation compared with chronic beta-alanine supplementation alone. Sodium bicarbonate ingestion led to increases in plasma pH, base excess, bicarbonate, and lactate concentrations. Both chronic beta-alanine and acute sodium bicarbonate supplementation alone had positive effects on 2,000-m rowing performance. The addition of acute sodium bicarbonate to chronic beta-alanine supplementation may further enhance rowing performance.

  8. Alanine synthesis from glyceraldehyde and ammonium ion in aqueous solution

    Science.gov (United States)

    Weber, A. L.

    1985-01-01

    The formation of alanine (ala) form C(14)-glyceraldehyde and ammonium phosphate in the presence or absence of a thiol is reported. At ambient temperature, ala synthesis was six times more rapid in the presence of 3-mercaptopropionic acid than in its absence (0.6 and 0.1 percent, respectively, after 60 days). Similarly, the presence of another thiol, N-acetylcysteinate, increased the production of ala, as well as of lactate. The reaction pathway of thiol-catalyzed synthesis of ala, with the lactic acid formed in a bypath, is suggested. In this, dehydration of glyceraldehyde is followed by the formation of hemithioacetal. In the presence of ammonia, an imine is formed, which eventually yields ala. This pathway is consistent with the observation that the rate ratio of ala/lactate remains constant throughout the process. The fact that the reaction takes place under anaerobic conditions in the presence of H2O and with the low concentrations of simple substrates and catalysts makes it an attractive model prebiotic reaction in the process of molecular evolution.

  9. Alanine/RPE dosimetry in the process of blood irradiation

    International Nuclear Information System (INIS)

    Chen, F.; Covas, D.T.; Baffa, O.

    2000-01-01

    A set of eighty dosemeters approximately of Dl- alanine was proportioned to the Hemo center of Hospital das Clinicas of Medicine Faculty at Ribeirao Preto (HC-FRMP) in the Sao Paulo University, with the purpose to realize a quality control of the radiation dose supplied to blood bags. These dosemeters were divided in eight groups and irradiated for a two months period.The dosemeters were adhered to the bags and arranged inside of an acrylic box filled water with dimensions: (40x40x20) cm 3 . The box irradiation was made using two fields parallel-opposed of (40x40) cm 2 at 80 cm distance source-surface, in the Radiotherapy Service of HC-FMRP, with a 60 Co teletherapy unit. The irradiation time was sufficient to supply a dose of 20 Gy approximately in the box center. The RPE measures were realized in a Varian E-4 spectrometer operating in X-band. For the total of dosemeters and for the irradiation volume, the minimum and maximum doses were of 14 Gy and 23 Gy respectively. The mean dose was (18±2) Gy (1 σ), and the variability coefficient 11 %. Around 5 % of the bags received a dose under of the recommended limit by the Brazilian legislation and as consequence, the exposure time was increased. (Author)

  10. The infrared spectrum of solid L-alanine: influence of pH-induced structural changes.

    Science.gov (United States)

    Garcia, Ana R; de Barros, Ricardo Brito; Lourenço, João P; Ilharco, Laura M

    2008-09-11

    The influence of the pH on the infrared spectrum of L-alanine has been analyzed by diffuse reflectance infrared Fourier transform (DRIFT) spectroscopy. The amino acid was precipitated from aqueous solutions and dried at 36.5 degrees C, in order to stabilize cationic L-alanine or alaninium [CH3CH(NH3(+))COOH] at pH 1, the zwitterionic form [CH3CH(NH3(+))COO(-)] at pH 6, and anionic L-alanine or alaninate [CH3CH(NH2)COO(-)] at pH 13. New insight on the specific inter and intramolecular interactions in the different forms of L-alanine was reached by a novel methodological approach: an infrared technique not used before to analyze solid amino acid samples (DRIFTS), in combination with a detailed analysis based on spectral deconvolution. The frequency ranges of interest include the carbonyl/carboxyl stretching and amine deformation modes and the OH/NH stretching modes. It was shown that intermolecular hydrogen bonds between the NH3(+) and COO(-) groups are predominant in the zwitterionic form, whereas in cationic L-alanine, H bonds between the COOH groups are responsible for the formation of dimers. In anionic L-alanine, only strong electrostatic interactions between the COO(-) groups and Na(+) ions are proposed, evidencing the relevant role of the counterion.

  11. Revised mechanism of d-alanine incorporation into cell wall polymers in Gram-positive bacteria

    Science.gov (United States)

    Reichmann, Nathalie T.; Cassona, Carolina Picarra

    2013-01-01

    Teichoic acids (TAs) are important for growth, biofilm formation, adhesion and virulence of Gram-positive bacterial pathogens. The chemical structures of the TAs vary between bacteria, though they typically consist of zwitterionic polymers that are anchored to either the peptidoglycan layer as in the case of wall teichoic acid (WTA) or the cell membrane and named lipoteichoic acid (LTA). The polymers are modified with d-alanines and a lack of this decoration leads to increased susceptibility to cationic antimicrobial peptides. Four proteins, DltA–D, are essential for the incorporation of d-alanines into cell wall polymers and it has been established that DltA transfers d-alanines in the cytoplasm of the cell onto the carrier protein DltC. However, two conflicting models have been proposed for the remainder of the mechanism. Using a cellular protein localization and membrane topology analysis, we show here that DltC does not traverse the membrane and that DltD is anchored to the outside of the cell. These data are in agreement with the originally proposed model for d-alanine incorporation through a process that has been proposed to proceed via a d-alanine undecaprenyl phosphate membrane intermediate. Furthermore, we found that WTA isolated from a Staphylococcus aureus strain lacking LTA contains only a small amount of d-alanine, indicating that LTA has a role, either direct or indirect, in the efficient d-alanine incorporation into WTA in living cells. PMID:23858088

  12. 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

  13. Alanine-EPR as a transfer standard dosimetry system for low energy X radiation

    International Nuclear Information System (INIS)

    Khoury, H.J.; Silva, E.J. da; Mehta, K.; Barros, V.S. de; Asfora, V.K.; Guzzo, P.L.; Parker, A.G.

    2015-01-01

    The purpose of this paper is to evaluate the use of alanine-EPR as a transfer standard dosimetry system for low energy X radiation, such as that in RS-2400, which operates in the range from 25 to 150 kV and 2 to 45 mA. Two types of alanine dosimeters were investigated. One is a commercial alanine pellets from Aérial-Centre de Ressources Technologiques, France and one was prepared in our laboratory (LMRI-DEN/UFPE). The EPR spectra of the irradiated dosimeters were recorded in the Nuclear Energy Department of UFPE, using a Bruker EMX10 EPR spectrometer operating in the X-band. The alanine-EPR dosimetry system was calibrated in the range of 20–220 Gy in this X-ray field, against an ionization chamber calibrated at the relevant X-ray energy with traceability to PTB. The results showed that both alanine dosimeters presented a linear dose response the same sensitivity, when the EPR signal was normalized to alanine mass. The total uncertainty in the measured dose was estimated to be about 3%. The results indicate that it is possible to use the alanine-EPR dosimetry system for validation of a low-energy X ray irradiator, such as RS-2400.

  14. 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)

  15. Knockout of the alanine racemase gene in Lactobacillus plantarum results in septation defects and cell wall perforation

    NARCIS (Netherlands)

    Palumbo, E.; Favier, C.F.; Deghorain, M.; Cocconcelli, P.S.; Grangette, C.; Mercenier, A.M.E.; Vaughan, E.E.; Hols, P.

    2004-01-01

    A stable mutant of Lactobacillus plantarum deficient in alanine racemase (Alr) was constructed by two successive homologous recombination steps. When the mutant was supplemented with D-alanine, growth and viability were unaffected. Surprisingly, deprivation Of D-alanine during exponential growth did

  16. Sensitivity comparison of two L-alanine doped blends to different photon energies

    International Nuclear Information System (INIS)

    Chen, Felipe; Vega Ramirez, Jose; Nicolucci, Patricia; Baffa, Oswaldo

    2008-01-01

    Full text: Blends of L-alanine (85% weight proportion) with KI (10%) and with PbI 2 (10%), these last two compounds acting as dopants, and with PVA (5%) acting as binder, were prepared in water at 80 C degrees. A blend of pure L-alanine (95%) with PVA (5%) was also prepared. The three blends were irradiated with photon beams of different energies (120 kV, 60 Co and 10 MV) with a unique dose of 30 Gy to compare their sensitivities for those three energies. EPR spectra of the three irradiated blends were recorded in a K-Band spectrometer (24 GHz) taking aliquots of about 4 mg for each blend. The energy sensitivity of a blend was defined as the peak-to-peak amplitude of its EPR spectrum central line. For the 60 Co energy (1.25 MeV) the blends presented practically the same sensitivity indicating that the presence of the dopants does not affect the sensitivity of L-alanine. For 10 MV X-rays there was an increment (around 20% - 30 %) in sensitivity for the two L-alanine doped blends compared with the pure L-alanine blend (not doped). In the case of 120 kV X-rays, the blends ala+KI and ala+PbI 2 showed an increment of 10 and 20 times, respectively, more sensitivity than the pure L-alanine blend. It is concluded that the dopants KI and PbI 2 produce a great enhance of the L-alanine sensitivity to low-energy photons. For the same dopant's content (10%) in the blend, PbI 2 showed a better performance. These results encourage us to try to enhance the sensitivity of L-alanine even more increasing the dopant's content in the blend. Application of these L-alanine doped blends in the dosimetry in diagnostic radiology could be possible. (author)

  17. d-Alanine metabolism is essential for growth and biofilm formation of Streptococcus mutans.

    Science.gov (United States)

    Qiu, W; Zheng, X; Wei, Y; Zhou, X; Zhang, K; Wang, S; Cheng, L; Li, Y; Ren, B; Xu, X; Li, Y; Li, M

    2016-10-01

    Part of the d-alanine (d-Ala) metabolic pathway in bacteria involves the conversion of l-alanine to d-Ala by alanine racemase and the formation of d-alanyl-d-alanine by d-alanine-d-alanine ligase, the product of which is involved in cell wall peptidoglycan synthesis. At present, drugs that target the metabolic pathway of d-Ala are already in clinical use - e.g. d-cycloserine (DCS) is used as an antibiotic against Mycobacterium tuberculosis. Streptococcus mutans is the main cariogenic bacterium in the oral cavity. Its d-Ala metabolism-associated enzymes alanine racemase and d-alanine-d-alanine ligase are encoded by the genes smu.1834 and smu.599, respectively, which may be potential targets for inhibitors. In this study, the addition of DCS blocked the d-Ala metabolic pathway in S. mutans, leading to bacterial cell wall defects, significant inhibition of bacterial growth and biofilm formation, and reductions in extracellular polysaccharide production and bacterial adhesion. However, the exogenous addition of d-Ala could reverse the inhibitory effect of DCS. Through the means of drug regulation, our study demonstrated, for the first time, the importance of d-Ala metabolism in the survival and biofilm formation of S. mutans. If the growth of S. mutans can be specifically inhibited by designing drugs that target d-Ala metabolism, then this may serve as a potential new treatment for dental caries. © 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  18. Mitochondrial defects associated with β-alanine toxicity: relevance to hyper-beta-alaninemia

    Science.gov (United States)

    Shetewy, Aza; Shimada-Takaura, Kayoko; Warner, Danielle; Jong, Chian Ju; Mehdi, Abu-Bakr Al; Alexeyev, Mikhail; Takahashi, Kyoko; Schaffer, Stephen W.

    2016-01-01

    Hyper-beta-alaninemia is a rare metabolic condition that results in elevated plasma and urinary β-alanine levels and is characterized by neurotoxicity, hypotonia, and respiratory distress. It has been proposed that at least some of the symptoms are caused by oxidative stress; however, only limited information is available on the mechanism of reactive oxygen species generation. The present study examines the hypothesis that β-alanine reduces cellular levels of taurine, which are required for normal respiratory chain function; cellular taurine depletion is known to reduce respiratory function and elevate mitochondrial superoxide generation. To test the taurine hypothesis, isolated neonatal rat cardiomyocytes and mouse embryonic fibroblasts were incubated with medium lacking or containing β-alanine. β-alanine treatment led to mitochondrial superoxide accumulation in conjunction with a decrease in oxygen consumption. The defect in β-alanine-mediated respiratory function was detected in permeabilized cells exposed to glutamate/malate but not in cells utilizing succinate, suggesting that β-alanine leads to impaired complex I activity. Taurine treatment limited mitochondrial superoxide generation, supporting a role for taurine in maintaining complex I activity. Also affected by taurine is mitochondrial morphology, as β-alanine-treated fibroblasts undergo fragmentation, a sign of unhealthy mitochondria that is reversed by taurine treatment. If left unaltered, β-alanine-treated fibroblasts also undergo mitochondrial apoptosis, as evidenced by activation of caspases 3 and 9 and the initiation of the mitochondrial permeability transition. Together, these data show that β-alanine mediates changes that reduce ATP generation and enhance oxidative stress, factors that contribute to heart failure. PMID:27023909

  19. Serum alanine aminotransferase levels, hematocrit rate and body weight correlations before and after hemodialysis session

    Directory of Open Access Journals (Sweden)

    Edmundo Pessoa Lopes

    2009-01-01

    Full Text Available PURPOSE: To evaluate alanine aminotransferase levels before and after a hemodialysis session and to correlate these values with the hematocrit rate and weight loss during hemodialysis. PATIENTS AND METHODS: The serum alanine aminotransferase levels, hematocrit rate and body weight were measured and correlated before and after a single hemodialysis session for 146 patients with chronic renal failure. An receiver operating characteristic (ROC curve for the serum alanine aminotransferase levels collected before and after hemodialysis was plotted to identify hepatitis C virus-infected patients. RESULTS: The mean weight loss of the 146 patients during hemodialysis was 5.3% (p < 0.001. The mean alanine aminotransferase levels before and after hemodialysis were 18.8 and 23.9 IU/, respectively, denoting a significant 28.1% increase. An equally significant increase of 16.4% in the hematocrit rate also occurred after hemodialysis. The weight loss was inversely correlated with the rise in both the alanine aminotransferase level (r = 0.3; p < 0.001 and hematocrit rate (r = 0.5; p < 0.001. A direct correlation was found between the rise in alanine aminotransferase levels and the hematocrit during the hemodialysis session (r = 0.4; p < 0.001. Based on the ROC curve, the upper limit of the normal alanine aminotransferase level should be reduced by 40% relative to the upper limit of normal if the blood samples are collected before the hemodialysis session or by 60% if blood samples are collected after the session. CONCLUSION: In the present study, significant elevations in the serum alanine aminotransferase levels and hematocrit rates occurred in parallel to a reduction in body weight after the hemodialysis session. These findings suggest that one of the factors for low alanine aminotransferase levels prior to hemodialysis could be hemodilution in patients with chronic renal failure.

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

    Science.gov (United States)

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

    1988-04-01

    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-14C]alanine, [3-3H]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 [14C]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.(ABSTRACT TRUNCATED AT 250 WORDS)

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

    Energy Technology Data Exchange (ETDEWEB)

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

    1988-04-01

    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-/sup 14/C)alanine, (3-/sup 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 (/sup 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.

  2. 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

  3. 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.

  4. The coupling of ω-transaminase and Oppenauer oxidation reactions via intra-membrane multicomponent diffusion – A process model for the synthesis of chiral amines

    DEFF Research Database (Denmark)

    Esparza-Isunza, T.; González-Brambila, M.; Gani, Rafiqul

    2015-01-01

    In this study we consider the theoretical coupling of an otherwise thermodynamically limited ω-transaminase reaction to an Oppenauer oxidation, in order to shift the equilibria of both reactions, with the aim of achieving a significant (and important) increase in the yield of the desired chiral...... sequentially in fed batch–batch mode. Both reactors are interconnected through a semi-permeable membrane, where multicomponent intra-membrane transport takes place by diffusion and viscous flow. The kinetic modeling of both reactions has been carried out and model simulations show that in this way...

  5. A thin alanine-polyethylene film dosimetry system with diffuse reflection spectrophotometric evaluation

    International Nuclear Information System (INIS)

    Zagorski, Z.P.; Rafalski, A.

    1995-01-01

    Characteristics of a new alanine dosimeter in the shape of a thin film, with the measurement of optical absorption of the CH 3 CHCOO - radical is described. That type of dosimeter, ALA/DRS (for diffuse reflection spectrophotometry) is compared, to an alanine dosimeter with EPR evaluation (ALA/EPR for short). In many respects the simple ALA/DRS version, as the alanine-polyethylene composite is superior. The paper shows the importance of the new experimental approach to free radical research in solid state radiation chemistry. (author). 7 refs., 3 figs

  6. Fluoropolymer coated alanine films treated by atmospheric pressure plasmas − In comparison with gamma irradiation

    DEFF Research Database (Denmark)

    Kusano, Yukihiro; Bardenshtein, Alexander; Morgen, Per

    2018-01-01

    Fluoropolymer coated alanine films are treated by a dielectric barrier discharge and a gliding arc at atmospheric pressure as well as with gamma irradiation. The film surfaces and the underlying bulk materials are characterized before and after each treatment. The fluorine content decreases...... and the oxygen content increases at the fluoropolymer surfaces, while deposition of specific plasma energies in the alanine films is detected by electron paramagnetic resonance spectroscopy, indicating that not only the fluoropolymer surfaces but also the bulk alanine materials are modified. Differences...

  7. Inducible l-Alanine Exporter Encoded by the Novel Gene ygaW (alaE) in Escherichia coli ▿

    Science.gov (United States)

    Hori, Hatsuhiro; Yoneyama, Hiroshi; Tobe, Ryuta; Ando, Tasuke; Isogai, Emiko; Katsumata, Ryoichi

    2011-01-01

    We previously isolated a mutant hypersensitive to l-alanyl-l-alanine from a non-l-alanine-metabolizing Escherichia coli strain and found that it lacked an inducible l-alanine export system. Consequently, this mutant showed a significant accumulation of intracellular l-alanine and a reduction in the l-alanine export rate compared to the parent strain. When the mutant was used as a host to clone a gene(s) that complements the dipeptide-hypersensitive phenotype, two uncharacterized genes, ygaW and ytfF, and two characterized genes, yddG and yeaS, were identified. Overexpression of each gene in the mutant resulted in a decrease in the intracellular l-alanine level and enhancement of the l-alanine export rate in the presence of the dipeptide, suggesting that their products function as exporters of l-alanine. Since ygaW exhibited the most striking impact on both the intra- and the extracellular l-alanine levels among the four genes identified, we disrupted the ygaW gene in the non-l-alanine-metabolizing strain. The resulting isogenic mutant showed the same intra- and extracellular l-alanine levels as observed in the dipeptide-hypersensitive mutant obtained by chemical mutagenesis. When each gene was overexpressed in the wild-type strain, which does not intrinsically excrete alanine, only the ygaW gene conferred on the cells the ability to excrete alanine. In addition, expression of the ygaW gene was induced in the presence of the dipeptide. On the basis of these results, we concluded that YgaW is likely to be the physiologically most relevant exporter for l-alanine in E. coli and proposed that the gene be redesignated alaE for alanine export. PMID:21531828

  8. In vivo dosimetry with L-alpha-alanine

    Energy Technology Data Exchange (ETDEWEB)

    Boey, R.; Van Der Velden, K. [Industriele Hogeschool van het Gemeenschapsonderwijs Limburg, Hasselt (Belgium); Schaeken, B. [Algemeen Ziekenhuis Middelheim, Antwerp (Belgium). Dept. of Radiotherapy

    1995-12-01

    When organic substances are irradiated, stable electrons can be formed. The concentration of these electrons is detected via electron paramagnetic resonance (EPR), a non-destructive form of dosimetry. L-alpha-alanine is extremely suited as a detector because of its high stability and high yield of unpaired electrons. With an EMS 104 spectrometer, we measure the peak-to-peak value of the first derivate of the resonance-spectrum. This value is proportional to the concentration of unpaired electrons and therefore with the absorbed dose. Prior to the in vivo measurements in teletherapy, a calibration curve had to be established. This clearly showed a linear relationship between the EPR-signal and the absorbed dose, except for very low dose where precision was low (20% 1 sd). This indicates that the background signal of the dosimeter is strongly orientation dependent. For this reason it was decided to use pre-irradiated detectors. A number of in vivo measurements has been performed. It was found that the error propagation plays a major role in the calculation of the measured absorbed dose, in the range 1 Gy-6 Gy. Contrary to in vivo measurements in brachytherapy, where higher doses are measured, large uncertainties (30% 1 sd) on the entry dose calculations were observed. For this reason, it is recommended to use a statistical method of reducing this standard deviation to an acceptable level. The proposed method, consisting of 2 detectors and the usage of weight coefficients on our standard deviations, gave promising results. However, theoretical calculations and in vivo measurements show that this method is still not satisfactory to reduce the uncertainty to an acceptable standard in clinical situations.

  9. Determination of the carbon, hydrogen and nitrogen contents of alanine and their uncertainties using the certified reference material L-alanine (NMIJ CRM 6011-a).

    Science.gov (United States)

    Itoh, Nobuyasu; Sato, Ayako; Yamazaki, Taichi; Numata, Masahiko; Takatsu, Akiko

    2013-01-01

    The carbon, hydrogen, and nitrogen (CHN) contents of alanine and their uncertainties were estimated using a CHN analyzer and the certified reference material (CRM) L-alanine. The CHN contents and their uncertainties, as measured using the single-point calibration method, were 40.36 ± 0.20% for C, 7.86 ± 0.13% for H, and 15.66 ± 0.09% for N; the results obtained using the bracket calibration method were also comparable. The method described in this study is reasonable, convenient, and meets the general requirement of having uncertainties ≤ 0.4%.

  10. Different β-alanine dimeric forms in trifluoromethanesulfonic acid salts. XRD and vibrational studies

    Science.gov (United States)

    Wołoszyn, Łukasz; Ilczyszyn, Maria M.

    2018-03-01

    Two new crystalline salts: β-alaninium trifluoromethanesulfonate (β-AlaOTf) and bis(β-alanine) trifluoromethanesulfonate (β-2AlaOTf) were obtained. The former one contains diprotonated β-alanine dimer, the latter one monoprotonated β-alanine dimer. Both compounds were studied by single crystal XRD, vibrational (IR and Raman) spectroscopy and calorimetric method. The quantum-mechanical calculations (DFT/B3LYP/6-311 ++G(2d,2p)) for the diprotonated dimer were carried out. The β-AlaOTf salt crystallizes in the P 1 bar space group of triclinic system (Z = 2), the β-2AlaOTf in the P21/m space group of monoclinic system (Z = 2). The vibrational data for the studied compounds are discussed in relation to their crystal structure, and provide insight into the character of hydrogen bonds and β-alanine protonation. The studied crystals do not exhibit phase transitions in the solid state.

  11. Titration of Alanine Monitored by NMR Spectroscopy: A Biochemistry Laboratory Experiment

    Science.gov (United States)

    Waller, Francis J.; And Others

    1977-01-01

    The experiment described here involves simultaneous monitoring of pH and NMR chemical shifts during an aqueous titration of alpha- and beta-alanine. This experiment is designed for use in an undergraduate biochemistry course. (MR)

  12. Dosimetric calibration of humidity chamber inside the 60Co-PANBIT irradiator using alanine ESR dosimeter

    International Nuclear Information System (INIS)

    Murali, S.; Venkataramani, R.; Pushparaja; Natarajan, V.; Sastry, M.D.; Bora, J.S.; Venkatacharyulu, K.

    1998-01-01

    In the present work the suitability of the electron spin resonance (ESR) technique, using DL - α - alanine sample is examined. It is well documented that radiation induced free radicals in alanine give strong ESR spectrum which can be used for dosimetry purposes. The aspects that are relevant to the present work are: (I) stability of the radicals in temperature and humidity conditions in which the experiments were carried out; (II) linearity at high doses; and (III) establishing the utility of alanine ESR dosimeter by cross checking the dose values with more established dosimeters. The details of these investigations are presented and it is shown that alanine ESR dosimeter will meet all the required conditions satisfactorily

  13. Preparation of alanine/ESR dosimeter using different binder of polymer blend

    International Nuclear Information System (INIS)

    Razzak, M.T.; Sudiro, Sutjipto; Sudradjat, Adjat; Waskito, Ashar; Djamili, M.F.

    1995-01-01

    Different composition of polymer blend of low density polyethylene (PE) and polystyrene (PS) have been studied to be used as a binder for the preparation of Alanine/ESR dosimeter. The polymer binder and Alanine powder were blended in Laboplastomil Mixer at 140 o C and then it was pressed into a plastic film of 0.50 mm thickness. The film was cut into sample size of 250 mm x 2.5 mm and irradiated by gamma rays from a cobalt-60 source at different dose and dose rate. It was found that a blend of Alanine, PS and PE in composition of 60:30:10 is appropriate to prepare the Alanine/ESR dosimeter. (author)

  14. 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

  15. Influence of convulsants on rat brain activities of alanine aminotransferase and aspartate aminotransferase

    Czech Academy of Sciences Publication Activity Database

    Netopilová, M.; Haugvicová, Renata; Kubová, Hana; Dršata, J.; Mareš, Pavel

    2001-01-01

    Roč. 26, č. 12 (2001), s. 1285-1291 ISSN 0364-3190 Institutional research plan: CEZ:AV0Z5011922 Keywords : alanine aminotransferase * aspartale aminotransferase Subject RIV: FH - Neurology Impact factor: 1.638, year: 2001

  16. Implementation of an alanine dosimetry service; Puesta en marcha de un servicio de dosimetria de alanima

    Energy Technology Data Exchange (ETDEWEB)

    Gago Arias, A.; Nunez Pelaez, N.; Peteiro Vilaseco, E.; Gomez Rodriguez, F.; Gonzalez Castano, D. M.

    2011-07-01

    This work facing the implementation of an alanine dosimetry service, linked to the installation of Co{sub 6}0 Radio physics Laboratory (LP) and Paramagnetic Resonance Service of the University of Santiago de Compostela (USC).

  17. Solid phase extraction of β-N-methylamino-L-alanine (BMAA) from ...

    African Journals Online (AJOL)

    2011-01-25

    N-methylamino-L-alanine, BMAA, cyanobacterial bloom, solid phase extraction. Introduction. The isolation of the neurotoxin BMAA from freshwater and marine cyanobacteria (Cox et al., 2005; Banack et al., 2007;. Esterhuizen ...

  18. Fish to meat intake ratio and cooking oils are associated with hepatitis C virus carriers with persistently normal alanine aminotransferase levels.

    Science.gov (United States)

    Otsuka, Momoka; Uchida, Yuki; Kawaguchi, Takumi; Taniguchi, Eitaro; Kawaguchi, Atsushi; Kitani, Shingo; Itou, Minoru; Oriishi, Tetsuharu; Kakuma, Tatsuyuki; Tanaka, Suiko; Yagi, Minoru; Sata, Michio

    2012-10-01

      Dietary habits are involved in the development of chronic inflammation; however, the impact of dietary profiles of hepatitis C virus carriers with persistently normal alanine transaminase levels (HCV-PNALT) remains unclear. The decision-tree algorithm is a data-mining statistical technique, which uncovers meaningful profiles of factors from a data collection. We aimed to investigate dietary profiles associated with HCV-PNALT using a decision-tree algorithm.   Twenty-seven HCV-PNALT and 41 patients with chronic hepatitis C were enrolled in this study. Dietary habit was assessed using a validated semiquantitative food frequency questionnaire. A decision-tree algorithm was created by dietary variables, and was evaluated by area under the receiver operating characteristic curve analysis (AUROC).   In multivariate analysis, fish to meat ratio, dairy product and cooking oils were identified as independent variables associated with HCV-PNALT. The decision-tree algorithm was created with two variables: a fish to meat ratio and cooking oils/ideal bodyweight. When subjects showed a fish to meat ratio of 1.24 or more, 68.8% of the subjects were HCV-PNALT. On the other hand, 11.5% of the subjects were HCV-PNALT when subjects showed a fish to meat ratio of less than 1.24 and cooking oil/ideal bodyweight of less than 0.23 g/kg. The difference in the proportion of HCV-PNALT between these groups are significant (odds ratio 16.87, 95% CI 3.40-83.67, P = 0.0005). Fivefold cross-validation of the decision-tree algorithm showed an AUROC of 0.6947 (95% CI 0.5656-0.8238, P = 0.0067).   The decision-tree algorithm disclosed that fish to meat ratio and cooking oil/ideal bodyweight were associated with HCV-PNALT. © 2012 The Japan Society of Hepatology.

  19. ESR/Alanine {gamma}-dosimetry in the 10-30 Gy range

    Energy Technology Data Exchange (ETDEWEB)

    Fainstein, C. E-mail: cfainstein@cab.cnea.gov.ar; Winkler, E.; Saravi, M

    2000-05-15

    We report Alanine Dosimeter preparation, procedures for using the ESR/Dosimetry method, and the resulting calibration curve for {gamma}-irradiation in the range from 10-30 Gy. We use calibration curve to measure the irradiation dose in {gamma}-irradiation of human blood, as required in Blood Transfusion Therapy. The ESR/Alanine results are compared against those obtained using the thermoluminescent dosimetry (TLD) method.

  20. Synthesis of racemic 2-deutero-3-fluoro-alanine and its salts, and intermediates therefor

    International Nuclear Information System (INIS)

    1975-01-01

    The racemates of 2-deutero-3-fluoro-alanine and its salts are prepared by reductive amination of 3-fluoro-pyruvic acid, its hydrate or salts thereof, via the intermediate 2-imino-3-fluoro propionic acid salt, using alkali metal borodeuterides as reducing agents. The racemates thus obtained are valuable in the production of the corresponding 2-deutero-3-fluoro-D-alanine, and its pharmacologically acceptable salts, and derivatives thereof, which are potent antibacterial agents

  1. 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.

  2. Alanine-EPR dosimetry system for high industrial as well radiotherapeutic dose measurement

    International Nuclear Information System (INIS)

    Dobrovodsky, J.; Bukovjan, J.

    2005-01-01

    Slovak Institute of Metrology is developing new metrology standard for high doses, based on the alanine-EPR as a reference dosimetry system. A Bruker e-scan EPR analyser developed specifically for alanine dosimetry has improved stability of EPR measurement, especially at lower dose range. The standard e-scan system provides sensitivity below 1 Gray. After further improvement of the system and lowering of dose determination expanded uncertainty down below 1 %, its utilisation for radiotherapy field is expected (authors)

  3. SU-E-T-643: Pure Alanine Dosimeter for Verification Dosimetry in IMRT

    International Nuclear Information System (INIS)

    Al-Karmi, Anan M.; Zraiqat, Fadi

    2015-01-01

    Purpose: The objective of this study was evaluation of accuracy of pure alanine dosimeters measuring intensity-modulated radiation therapy (IMRT) dose distributions in a thorax phantom. Methods: Alanine dosimeters were prepared in the form of 110 mg pure L-α-alanine powder filled into clear tissue-equivalent polymethylmethacrylate (PMMA) plastic tubes with the dimensions 25 mm length, 3 mm inner diameter, and 1 mm wall thickness. A dose-response calibration curve was established for the alanine by placing the dosimeters at 1.5 cm depth in a 30×30×30 cm 3 solid water phantom and then irradiating on a linac with 6 MV photon beam at 10×10 cm 2 field size to doses ranging from 1 to 5 Gy. Electron paramagnetic resonance (EPR) spectroscopy was used to determine the absorbed dose in alanine. An IMRT treatment plan was designed for a commercial heterogeneous CIRS thorax phantom and the dose values were calculated at three different points located in tissue, lung, and bone equivalent materials. A set of dose measurements was carried out to compare measured and calculated dose values by placing the alanine dosimeters at those selected locations inside the thorax phantom and delivering the IMRT to the phantom. Results: The alanine dose measurements and the IMRT plan dose calculations were found to be in agreement within ±2%. Specifically, the deviations were −0.5%, 1.3%, and −1.7% for tissue, lung, and bone; respectively. The slightly large deviations observed for lung and bone may be attributed to tissue inhomogeneity, steep dose gradients in these regions, and uncontrollable changes in spectrometer conditions. Conclusion: The results described herein confirmed that pure alanine dosimeter was suitable for in-phantom dosimetry of IMRT beams because of its high sensitivity and acceptable accuracy. This makes the dosimeter a promising option for quality control of the therapeutic beams, complementing the commonly used ionization chambers, TLDs, and films

  4. Use of a range scaling method to determine alanine/water stopping power ratios

    International Nuclear Information System (INIS)

    McEwen, M.R.; Sephton, J.P.; Sharpe, P.H.G.; Shipley, D.R.

    2003-01-01

    A phantom composed of alanine dosimeter material has been constructed and depth-dose measurements made in a 10 MeV electron beam. The results have demonstrated the feasibility of using relative depth-dose measurements to determine stopping power ratios in materials of dosimetric interest. Experimental stopping power ratios for alanine dosimeter material and water agreed with the data of ICRU Report 37 within the uncertainty of the experiment (±1.2% at a 95% confidence level)

  5. No Effect of β-alanine on Muscle Function and Kayak Performance.

    Science.gov (United States)

    Bech, Signe Refsgaard; Nielsen, Tobias Schmidt; Hald, Martin; Jakobsen, Jarl Pors; Nordsborg, Nikolai Baastrup

    2018-03-01

    If β-alanine supplementation counteracts muscular fatigue development or improves athletic performance was investigated. Elite kayak rowers (10 men and 7 women) were supplemented with either 80 mg·kg body mass·d of β-alanine or placebo for 8 wk. Muscular fatigue development was investigated by applying a 2-min elbow flexor maximal voluntary contraction (MVC). EMG was recorded continuously, and voluntary activation was determined 30, 60, 90, and 115 s into the 2-min MVC. In addition, performance was evaluated as 1000-m and 5 × 250-m kayak ergometer rowing. Force reduction during the 2-min MVC was similar before and after supplementation with β-alanine (30.9% ± 10.3% vs 36.0% ± 14.1%) and placebo (35.5% ± 7.7% vs 35.1% ± 8.0%). No time effect was apparent in voluntary activation during the 2-min MVC. In addition, there was no detectable effect of β-alanine supplementation on 1000-m kayak ergometer performance (β-alanine: 0.26% ± 0.02% vs placebo: -0.18% ± 0.02%) or accumulated 5 × 250-m time (β-alanine: -1.0% ± 0.3% vs placebo: -1.0% ± 0.2%). In 5 × 250 m, mean power output was reduced to a similar extent from first to fifth interval before and after supplementation with β-alanine (23% ± 11% vs 22% ± 10%) and placebo (26% ± 13% vs 20% ± 5%). Two-minute MVC characteristics are unaffected by β-alanine supplementation in elite kayakers, and likewise, both a 1000-m kayak ergometer time trial lasting 4-5 min and a 5 × 250-m repeated sprint ability were unaltered by supplementation.

  6. A facile and efficient synthesis of (Purin-6-yl)alanines.

    Science.gov (United States)

    Capek, Petr; Pohl, Radek; Hocek, Michal

    2004-11-12

    (Purin-6-yl)alanines, a new class of amino acid-nucleobase conjugates, were synthesized by palladium-catalyzed cross-coupling reactions of protected iodozincalanines with 6-iodopurines (9-Bn-6-iodopurine and 9-THP-6-iodopurine as well as acyl-protected 6-iodopurine ribonucleoside and 2-deoxyribonucleoside). Free purine base and nucleosides bearing alanine in position 6 were obtained after complete deprotection of the products of cross-coupling reactions.

  7. Eukaryotic beta-alanine synthases are functionally related but have a high degree of structural diversity

    DEFF Research Database (Denmark)

    Gojkovic, Zoran; Sandrini, Michael; Piskur, Jure

    2001-01-01

    activity was used to clone analogous genes from different eukaryotes. Putative PYD3 sequences from the yeast S. kluyveri, the slime mold Dictyostelium discoideum, and the fruit fly Drosophila melanogaster complemented the pyd3 defect. When the S. kluyveri PYD3 gene was expressed in S. cerevisiae, which has......-carbamyl-beta -alanine, but not by uracil. This wrork establishes S. kluyveri as a model organism for studying pyrimidine degradation and beta -alanine production in eukaryotes....

  8. The influence of measurement and storage conditions on alanine ESR dosimeters

    International Nuclear Information System (INIS)

    Alexandre, A.C.

    1992-01-01

    Alanine has several desirable properties as an ESR dosemeter e.g. tissue equivalence, low fading and an approximately linear response for doses up to 10 kGy. This work reports on a simple system to produce the alanine dosemeter, the signal intensity for a range of doses and energies, and the effect of the air humidity and the spectrometer settings on the ESR signal. (Author)

  9. EPR dosimetry of radiotherapy photon beams in inhomogeneous media using alanine films

    International Nuclear Information System (INIS)

    Oesteraas, Bjoern Helge; Hole, Eli Olaug; Olsen, Dag Rune; Malinen, Eirik

    2006-01-01

    In the current work, EPR (electron paramagnetic resonance) dosimetry using alanine films (134 μm thick) was utilized for dose measurements in inhomogeneous phantoms irradiated with radiotherapy photon beams. The main phantom material was PMMA, while either Styrofoam or aluminium was introduced as an inhomogeneity. The phantoms were irradiated to a maximum dose of about 30 Gy with 6 or 15 MV photons. The performance of the alanine film dosimeters was investigated and compared to results from ion chamber dosimetry, Monte Carlo simulations and radiotherapy treatment planning calculations. It was found that the alanine film dosimeters had a linear dose response above approximately 5 Gy, while a background signal obscured the response at lower dose levels. For doses between 5 and 60 Gy, the standard deviation of single alanine film dose estimates was about 2%. The alanine film dose estimates yielded results comparable to those from the Monte Carlo simulations and the ion chamber measurements, with absolute differences between estimates in the order of 1-15%. The treatment planning calculations exhibited limited applicability. The current work shows that alanine film dosimetry is a method suitable for estimating radiotherapeutical doses and for dose measurements in inhomogeneous media

  10. Accurate measurement of the optical activity of alanine crystals and the determination of their absolute chirality

    Science.gov (United States)

    Ishikawa, Kazuhiko; Terasawa, Yukana; Tanaka, Masahito; Asahi, Toru

    2017-05-01

    Wavelength dependence measurements of the chiroptical properties in alanine crystals have so far been unsuccessful using conventional spectroscopic techniques. We describe our attempts to measure the wavelength dependence of the optical activity in L- and D-alanine crystals along each crystallographic axis, and to determine the absolute chirality of alanine crystals by correlating the absolute structure to the optical activity using an x-ray diffractometer and a generalized high accuracy universal polarimeter. We have succeeded in accurately measuring the optical rotatory dispersion in the direction, which shows that the optical rotation of the D-alanine crystal is dextrorotatory and that of the L-alanine crystal is laevorotatory, thereby determining the absolute chirality. Furthermore, comparison with the optical activity in solution shows that the optical activity in alanine crystals is different not only in value, but also in the sign. These results have led us to conclude that the optical rotatory power in the crystalline state should not be simply the summation of molecular optical rotatory power values. We propose the necessity of a theory, which contains the contribution of molecular interactions within the crystal, in order to calculate the optical rotatory power of the crystalline state.

  11. Dose intercomparison study involving Fricke, ethanol chlorobenzene, PMMA and alanine dosimeters

    International Nuclear Information System (INIS)

    Lanuza, L.G.; Cabalfin, E.G.; Kojima, T.; Tachibana, H.

    1999-01-01

    A dose intercomparison study was carried out between the Philippine Nuclear Research Institute (PNRI) and Takasaki Radiation Chemistry Research Establishment, Japan Atomic Energy Research Institute (JAERI) to determine reliability of the dosimetry systems being used by PNRI employing ethanol chlorobenzene (ECB), Fricke and alanine dosimeters. The Fricke and ECB dosimeters were prepared at PNRI while the alanine-polystyrene dosimeter was provided by JAERI. Fricke or ECB dosimeters were irradiated together with alanine at PNRI gamma irradiation facilities. Analyses of the Fricke and ECB dosimeters were performed at PNRI while alanine dosimeters were analyzed at JAERI. A comparison study between alanine and polymethylmethacrylate (PMMA, Radix RN15) dosimeters was also undertaken at JAERI. The dosimeters were irradiated together under different irradiation conditions using the gamma irradiation facilities of JAERI and Radia Industry Co. Ltd. (Japan). Evaluations of PMMA and alanine dosimeters were both performed at JAERI. Result of the dose intercomparison of PNRI with the International Atomic Energy Agency through the International Dose Assurance Service (IDAS) is also presented. (author)

  12. [Alanine solution as enzyme reaction buffer used in A to O blood group conversion].

    Science.gov (United States)

    Li, Su-Bo; Zhang, Xue; Zhang, Yin-Ze; Tan, Ying-Xia; Bao, Guo-Qiang; Wang, Ying-Li; Ji, Shou-Ping; Gong, Feng; Gao, Hong-Wei

    2014-06-01

    The aim of this study was to investigate the effect of alanine solution as α-N-acetylgalactosaminidase enzyme reaction buffer on the enzymatic activity of A antigen. The binding ability of α-N-acetylgalactosaminidase with RBC in different reaction buffer such as alanine solution, glycine solution, normal saline (0.9% NaCl), PBS, PCS was detected by Western blot. The results showed that the efficiency of A to O conversion in alanine solution was similar to that in glycine solution, and Western blot confirmed that most of enzymes blinded with RBC in glycine or alanine solution, but few enzymes blinded with RBC in PBS, PCS or normal saline. The evidences indicated that binding of enzyme with RBC was a key element for A to O blood group conversion, while the binding ability of α-N-acetylgalactosaminidase with RBC in alanine or glycine solution was similar. It is concluded that alanine solution can be used as enzyme reaction buffer in A to O blood group conversion. In this buffer, the α-N-acetylgalactosaminidase is closely blinded with RBC and α-N-acetylgalactosaminidase plays efficient enzymatic activity of A antigen.

  13. Dynamics of solid alanine by means of nuclear magnetic resonance relaxometry

    Science.gov (United States)

    Kubica-Misztal, A.; Rochowski, P.; Florek-Wojciechowska, M.; Kruk, D.

    2017-04-01

    1H nuclear magnetic resonance relaxometry was applied to investigate the dynamics of l-alanine in the solid phase (powder). The experimental studies were carried out in a very broad frequency range, covering four orders of magnitude—from 4 kHz to 40 MHz (referring to the 1H resonance frequency) in order to probe motional processes of much different time scales by a single experiment. To get access to the dynamics of different proton groups of alanine, the 1H spin-lattice relaxation measurements were performed for non-deuterated and partially deuterated alanine. The experiments were carried out in the temperature range of 293 K-370 K (non-deuterated alanine) and 318 K-370 K (partially deuterated alanine). As a result of a thorough theoretical analysis of the extensive set of experimental results, three motional processes occurring on different time scales are identified and quantitatively described. The slowest process occurs on a time scale of μs and it is attributed to the collective dynamics of a 3D hydrogen bond network of alanine, while the intermediate, attributed to the dynamics of the NH3 group, corresponds to the range of tenths of ns. The fast process describes the rotation of the CH3 group.

  14. 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

  15. Synthesis of silver nanoparticles using DL-alanine for ESR dosimetry applications

    International Nuclear Information System (INIS)

    Guidelli, Eder José; Ramos, Ana Paula; Zaniquelli, Maria Elisabete D.; Nicolucci, Patricia; Baffa, Oswaldo

    2012-01-01

    The potential use of alanine for the production of nanoparticles is presented here for the first time. Silver nanoparticles were synthesized using a simple green method, namely the thermal treatment of silver nitrate aqueous solutions with DL-alanine. The latter compound was employed both as a reducing and a capping agent. Particles with average size equal to 7.5 nm, face-centered cubic crystalline structure, narrow size distribution, and spherical shape were obtained. Interaction between the silver ions present on the surface of the nanoparticles and the amine group of the DL-alanine molecule seems to be responsible for reduction of the silver ions and for the stability of the colloid. The bio-hybrid nano-composite was used as an ESR dosimeter. The amount of silver nanoparticles in the nanocomposite was not sufficient to cause considerable loss of tissue equivalency. Moreover, the samples containing nanoparticles presented increased sensitivity and reduced energetic dependence as compared with pure DL-alanine, contributing to the construction of small-sized dosimeters. - Highlights: ► The synthesis is environmentally benign, easy to perform, and of low-cost. ► DL-Alanine was employed both as reducing and capping agent. ► Mean size of 7.5 nm, narrow size distribution, and spherical shape of particles. ► Increased sensitivity and reduced energetic dependence compared with pure alanine. ► The nanocomposite has potential application for ESR dosimetry.

  16. Alanine aminotransferase, gamma-glutamyltransferase (GGT) and all-cause mortality: results from a population-based Danish twins study alanine aminotransferase, GGT and mortality in elderly twins

    DEFF Research Database (Denmark)

    Fraser, Abigail; Thinggaard, Mikael; Christensen, Kaare

    2009-01-01

    Abstract Background/Aims: Alanine aminotransferase (ALT) and gamma-glutamyltransferase (GGT) are widely used markers of liver disease. Several population-based cohort studies have found associations of these liver enzymes with all-cause mortality. None of these studies controlled for genetic...

  17. Relationship between turnover rate and oxidation rate of alanine in the post-absorptive state and during parenteral nutrition before and after surgery

    NARCIS (Netherlands)

    Sauerwein, H. P.; Michels, R. P.; Cejka, V.

    1985-01-01

    The influence of total parenteral nutrition and stomach resection on alanine turnover rate and alanine oxidation rate was measured in ten patients after single injection of U-14 C-alanine. Sequential studies were done in three patients. During parenteral nutrition alanine turnover was significantly

  18. Effects of beta-alanine supplementation and interval training on physiological determinants of severe exercise performance.

    Science.gov (United States)

    Gross, Micah; Boesch, Chris; Bolliger, Christine S; Norman, Barbara; Gustafsson, Thomas; Hoppeler, Hans; Vogt, Michael

    2014-02-01

    We aimed to manipulate physiological determinants of severe exercise performance. We hypothesized that (1) beta-alanine supplementation would increase intramuscular carnosine and buffering capacity and dampen acidosis during severe cycling, (2) that high-intensity interval training (HIT) would enhance aerobic energy contribution during severe cycling, and (3) that HIT preceded by beta-alanine supplementation would have greater benefits. Sixteen active men performed incremental cycling tests and 90-s severe (110 % peak power) cycling tests at three time points: before and after oral supplementation with either beta-alanine or placebo, and after an 11-days HIT block (9 sessions, 4 × 4 min), which followed supplementation. Carnosine was assessed via MR spectroscopy. Energy contribution during 90-s severe cycling was estimated from the O2 deficit. Biopsies from m. vastus lateralis were taken before and after the test. Beta-alanine increased leg muscle carnosine (32 ± 13 %, d = 3.1). Buffering capacity and incremental cycling were unaffected, but during 90-s severe cycling, beta-alanine increased aerobic energy contribution (1.4 ± 1.3 %, d = 0.5), concurrent with reduced O2 deficit (-5.0 ± 5.0 %, d = 0.6) and muscle lactate accumulation (-23 ± 30 %, d = 0.9), while having no effect on pH. Beta-alanine also enhanced motivation and perceived state during the HIT block. There were no between-group differences in adaptations to the training block, namely increased buffering capacity (+7.9 ± 11.9 %, p = 0.04, d = 0.6, n = 14) and glycogen storage (+30 ± 47 %, p = 0.04, d = 0.5, n = 16). Beta-alanine did not affect buffering considerably, but has beneficial effects on severe exercise metabolism as well as psychological parameters during intense training phases.

  19. Studies of the uptake and release of [3H]β-alanine by frog spinal slices

    International Nuclear Information System (INIS)

    Adair, R.; Davidoff, R.A.

    1977-01-01

    [ 3 H]β-alanine was accumulated by frog spinal cord slices by two transport components with estimated Ksub(m) values of μM ('high-affinity') and 11 mM ('low affinity') respectively. The high affinity uptake exhibited sodium ion and energy dependence, temperature sensitivity, had a very low Vsub(max) (10.4 nmol/g/min) compared to GABA and glycine, was competitively inhibited by GABA (Ksub(i) 2μM), and was significantly reduced by the presence of glycine and of taurine in the incubating medium. When slices preloaded with [ 3 H]β-alanine were superfused with medium containing depolarizing concentrations of potassium ions, there was a small, but consistent, increase in [ 3 H]β-alanine efflux: 1.4 times prestimulation rates in 40 mM potassium. When the superfusate was altered by omission of calcium and addition of concentrations of magnesium (10 mM), manganese (1 mM), and cobalt (1 mM) ions sufficient to block reflex transmission in the isolated in vitro frog cord, the potassium-evoked release was not blocked. Release was decreased by lanthanum ions (1 mM). Release of [ 3 H]GABA and [ 3 H]glycine in parallel experiments was inhibited by magnesium, manganese, cobalt and lanthanum. Veratridine significantly increased the release of [ 3 H]GABA and [ 3 H]glycine but not of [ 3 H]β-alanine. These observations demonstrate the non-specificity of β-alanine uptake and the unconventional nature of the calcium-dependence of β-alanine release and therefore do not lend support to the hypothesis that β-alanine functions as a neurotransmitter in frog spinal cord. (author)

  20. Electron Paramagnetic Resonance signal ratio of Alanine Pellets In Nuclear Power Plants

    International Nuclear Information System (INIS)

    Choi, Hoon; Sung, In Bok; Lee, Byung Il; Lim, Young Ki

    2011-01-01

    As a dosimeter for ESR dosimetry, alanine has many useful features including relatively long endurance time of radicals and almost no difference with the radiation dose rate and radiation quality. Alanine dosimeters have been accepted as transfer dosimeters for their good precision at high radiation range. For alanine/ESR spectra, it was reported that the peaks of the spectra are due to the three kinds of radicals induced by radiation. The ratio of the weak 'satellite line' and the central peak of the three man in alanine specta(in this study x /y ratio ) are changed on the LET value of radiation. In case of lithium formate monohydrate was recently identified as a promising dosimetric material. in addition, it was reported the the peak height in the signal spectra is not easily saturated when power is irradiated with relatively high LET radiation such as neutron rays. The difference in the peak height ratio was reported to be caused by increased local radical density following the radiation of high LET. The spectrum shape of some alanine dosimeter installed in the containment buildings of NPPs showed differences in comparison with dosimeters exposed only to gamma rays. There was apparent change of spectra, expressed as the 'x/y ratio'. As noted in other papers, high LET radiation such as neutron rays causes shape changes of the spectrum of alanine dosimeters. Thus, the unanticipated high dose level and low 'x/y ratio' of some alanine dosimeters from the containment building could be explained b exposure to mixed radiation with high LET. Generally, the locations evaluated in this study are regarded as being exposed only to gamma rays, because the positions are blocked from direct neutron rays, because the positions are blocked from direct neutron rays from reactor by thick cement barriers and heavy instruments.

  1. Mechanism of leukotriene D4 inhibition of Na-alanine cotransport in intestinal epithelial cells.

    Science.gov (United States)

    Talukder, Jamilur R; Kekuda, Ramesh; Saha, Prosenjit; Sundaram, Uma

    2008-07-01

    In a rabbit model of chronic intestinal inflammation, we previously demonstrated inhibition of neutral Na-amino acid cotransport. The mechanism of the inhibition was secondary to a decrease in the affinity for amino acid rather than the number of cotransporters. Since leukotriene (LT)D4 is known to be elevated in enterocytes during chronic intestinal inflammation, we used rat intestinal epithelial cell (IEC-18) monolayers to determine the mechanism of regulation of Na-alanine cotransport (alanine, serine, cysteine transporter 1: ASCT1) by LTD4. Na-alanine cotransport was inhibited by LTD4 in IEC-18 cells. The mechanism of inhibition of ASCT1 (solute carrier, SLC1A4) by LTD4 is secondary to a decrease in the affinity of the cotransporter for alanine without a significant change in cotransporter numbers and is not secondary to an alteration in the Na+ extruding capacity of the cells. Real-time quantitative PCR and Western blot analysis results indicate that ASCT1 message and protein levels are also unchanged in LTD4-treated IEC-18 cells. These results indicate that LTD4 inhibits Na-dependent neutral amino acid cotransport in IEC. The mechanism of inhibition is secondary to a decrease in the affinity for alanine, which is identical to that seen in villus cells from the chronically inflamed rabbit small intestine, where LTD4 levels are significantly increased.

  2. L-alanine-induced germination in Bacillus licheniformis -the impact of native gerA sequences.

    Science.gov (United States)

    Madslien, Elisabeth H; Granum, Per Einar; Blatny, Janet M; Lindbäck, Toril

    2014-04-22

    L-alanine, acting through the GerA receptor, was recently found to be an efficient germinant in Bacillus licheniformis ATCC14580/DSM13. In this study, we show that several of 46 examined B. licheniformis strains germinate remarkably slower than the type strain when exposed to L-alanine. These strains are not necessarily closely related, as determined by MLST (multi-locus sequence typing). Three of the slow-germinating strains were further examined in order to see whether nucleotide substitutions in the gerA sequences were responsible for the slow L-alanine germination. This was performed by complementing the transformable type strain derivate MW3ΔgerAA with gerA variants from the three slow-germinating strains; NVH1032, NVH1112 and NVH800. A wide selection of B. licheniformis strains was evaluated for L-alanine-induced germination efficiency. Our results show that gerA substitutions could only partially explain why spores of some B. licheniformis strains responded slower than others in the presence of L-alanine.

  3. Optical signal response pf the alanine gel solution for photons and electrons clinical beams

    International Nuclear Information System (INIS)

    Silva, Cleber Feijo; Campos, Leticia Lucente

    2009-01-01

    Alanine gel dosimeter is a new gel material developed at IPEN that presents significant improvement on previous alanine systems developed by Costa (1994). The measure technique is based on the transformation of ferrous ions (Fe 2+ ) in ferric ions (Fe 3+ ) after irradiation. The DL-Alanine (C 3 H 7 NO 2 ) is an aminoacid tissue equivalent that improves the production of ferric ions in the solution. This work aims to study the comparison of optical signal response of the alanine gel solution for photons and electrons clinical beams. It was observed that the calibration factor can be considered independent of quality of the radiation for photons and electrons clinical beams. Therefore, it can be used the same calibration factor for evaluating the absorbed dose in photons and electrons fields in the energy of 6 MeV. Alanine Gel Dosimeter presents good performance and can be useful as alternative dosimeter in the radiotherapy area using MRI technique for 3D dose distribution evaluation. (author)

  4. ald of Mycobacterium tuberculosis Encodes both the Alanine Dehydrogenase and the Putative Glycine Dehydrogenase

    Science.gov (United States)

    Giffin, Michelle M.; Modesti, Lucia; Raab, Ronald W.; Wayne, Lawrence G.

    2012-01-01

    The putative glycine dehydrogenase of Mycobacterium tuberculosis catalyzes the reductive amination of glyoxylate to glycine but not the reverse reaction. The enzyme was purified and identified as the previously characterized alanine dehydrogenase. The Ald enzyme was expressed in Escherichia coli and had both pyruvate and glyoxylate aminating activities. The gene, ald, was inactivated in M. tuberculosis, which resulted in the loss of all activities. Both enzyme activities were found associated with the cell and were not detected in the extracellular filtrate. By using an anti-Ald antibody, the protein was localized to the cell membrane, with a smaller fraction in the cytosol. None was detected in the extracellular medium. The ald knockout strain grew without alanine or glycine and was able to utilize glycine but not alanine as a nitrogen source. Transcription of ald was induced when alanine was the sole nitrogen source, and higher levels of Ald enzyme were measured. Ald is proposed to have several functions, including ammonium incorporation and alanine breakdown. PMID:22210765

  5. Alanine Counteracts the Destabilizing Effect that Urea has on RNase-A.

    Science.gov (United States)

    Chowhan, Rimpy K; Ali, Fasil; Bhat, Mohd Y; Rahman, Safikur; Singh, Laishram R; Ahmad, Faizan; Dar, Tanveer A

    2016-01-01

    It is generally believed that organisms use and accumulate methylamine osmolytes to prevent urea's damaging effect on protein stability and activity. However, urea-rich cells not only accumulate methylamines but also many other methylated and non-methylated compounds as well. But, so far it is not known whether osmolytes that are not accumulated in urea-rich cells could also confer urea-counteracting properties. We investigated the behavior of a non-methylamine osmolyte, alanine for its counteracting effect against urea denaturation of a model protein, ribonuclease A (RNase-A). We have measured structure and thermodynamic parameters (Tm, ΔHm, and ΔGD°) of RNase-A in the presence of alanine, urea and their combination. The results were also compared with the ability of glycine (osmolyte lacking one methyl group when compared with alanine) to counter urea's effect on protein stability. We observed that alanine but not glycine counteracts urea's harmful effect on RNase-A stability. The results indicated that alanine (in addition to methylamine osmolytes) may serve as an alternate urea-counteractant. Since glycine fails to protect RNase-A from urea's destabilizing effect, it seems that methylation to glycine might have some evolutionary significance to protect proteins against harmful effects of urea.

  6. Characterization of lithium formate EPR dosimeters for high dose applications – comparison with alanine

    DEFF Research Database (Denmark)

    Waldeland, Einar; Helt-Hansen, Jakob; Malinen, Eirik

    2011-01-01

    Lithium formate and l-α-alanine (alanine) EPR dosimeters were irradiated to doses from 100 Gy to 100 kGy. The irradiations were mainly performed at a Gammacell irradiator with dose rate of approximately 5.5 kGy h−1. Both the peak-to-peak amplitude of the first derivative EPR spectrum and the area...... irradiated to 1 kGy at temperatures from 11ºC to 40ºC were analyzed. By fitting an ‘exponential rise to maximum'-function to the dependence of the area under the EPR absorption spectrum on the dose, saturation doses of 53 kGy and 87 kGy for lithium formate and alanine, respectively, were found. Lower...... signal per ºC change in irradiation temperature) were 0.154 % K−1 and 0.161 % K−1 for lithium formate and alanine, respectively. This work demonstrates that lithium formate EPR dosimeters may be suitable for high dose applications, but their signals saturate at lower doses than alanine. The saturation...

  7. Perturbation correction for alanine dosimeters in different phantom materials in high-energy photon beams.

    Science.gov (United States)

    von Voigts-Rhetz, P; Anton, M; Vorwerk, H; Zink, K

    2016-02-07

    In modern radiotherapy the verification of complex treatments plans is often performed in inhomogeneous or even anthropomorphic phantoms. For dose verification small detectors are necessary and therefore alanine detectors are most suitable. Though the response of alanine for a wide range of clinical photon energies in water is well know, the knowledge about the influence of the surrounding phantom material on the response of alanine is sparse. Therefore we investigated the influence of twenty different surrounding/phantom materials for alanine dosimeters in clinical photon fields via Monte Carlo simulations. The relative electron density of the used materials was in the range [Formula: see text] up to 1.69, covering almost all materials appearing in inhomogeneous or anthropomorphic phantoms used in radiotherapy. The investigations were performed for three different clinical photon spectra ranging from 6 to 25 MV-X and Co-60 and as a result a perturbation correction [Formula: see text] depending on the environmental material was established. The Monte Carlo simulation show, that there is only a small dependence of [Formula: see text] on the phantom material and the photon energy, which is below  ±0.6%. The results confirm the good suitability of alanine detectors for in-vivo dosimetry.

  8. Absorbed dose determination in kilovoltage X-ray synchrotron radiation using alanine dosimeters.

    Science.gov (United States)

    Butler, D J; Lye, J E; Wright, T E; Crossley, D; Sharpe, P H G; Stevenson, A W; Livingstone, J; Crosbie, J C

    2016-12-01

    Alanine dosimeters from the National Physical Laboratory (NPL) in the UK were irradiated using kilovoltage synchrotron radiation at the imaging and medical beam line (IMBL) at the Australian Synchrotron. A 20 × 20 mm 2 area was irradiated by scanning the phantom containing the alanine through the 1 mm × 20 mm beam at a constant velocity. The polychromatic beam had an average energy of 95 keV and nominal absorbed dose to water rate of 250 Gy/s. The absorbed dose to water in the solid water phantom was first determined using a PTW Model 31014 PinPoint ionization chamber traceable to a graphite calorimeter. The alanine was read out at NPL using correction factors determined for 60 Co, traceable to NPL standards, and a published energy correction was applied to correct for the effect of the synchrotron beam quality. The ratio of the doses determined by alanine at NPL and those determined at the synchrotron was 0.975 (standard uncertainty 0.042) when alanine energy correction factors published by Waldeland et al. (Waldeland E, Hole E O, Sagstuen E and Malinen E, Med. Phys. 2010, 37, 3569) were used, and 0.996 (standard uncertainty 0.031) when factors by Anton et al. (Anton M, Büermann L., Phys Med Biol. 2015 60 6113-29) were used. The results provide additional verification of the IMBL dosimetry.

  9. ESR investigation of L-α-alanine and sucrose radicals produced by heavy-ion irradiation

    International Nuclear Information System (INIS)

    Nakagawa, K.; Sato, Y.

    2005-01-01

    We investigated sucrose and L-α-alanine radicals produced by heavy (particle) ion irradiation with various LETs (linear energy transfer). The impact of the heavy ions on the samples produced stable free radicals, which were analyzed by ESR (electron spin resonance). Identical spectra were measured after one year. The obtained spectral patterns were the same as those for helium (He), carbon (C), and neon (Ne) ions irradiation. The absorbed dose dependences for the irradiated sucrose and alanine samples were examined. The ESR response has a linear relation with the absorbed dose. The ESR response at 60 Gy was slightly lower than a linear line for sucrose; however, the response showed good linearity for the alanine. In addition, the total spin concentration obtained by heavy-ion irradiation correlated logarithmically with the LET. Qualitative ESR analyse showed that the production of sucrose and alanine radicals depended on both different particle irradiation and the LET under the same dose. Thus, the present ESR results imply that sucrose together with L-α-alanine can be used to monitor LET as well as the number of ionizing particle for the production of stable free radicals. (author)

  10. Development of portable ESR spectrometer as a reader for alanine dosimeters

    International Nuclear Information System (INIS)

    Kojima, T.; Haruyama, Y.; Tachibana, H.; Tanaka, R.; Okamoto, J.

    1993-01-01

    A prototype portable electron spin resonance (ESR) spectrometer was designed and tested, and its feasibility as a reader of alanine dosimeters was studied from the two standpoints of reproducibility of readings and sensitivity sufficient for dosimetry in the absorbed dose range 1-100 kGy. It has two main components: a permanent magnet and resonator; and a unit box with a microwave and auto-frequency control (AFC) circuit, a sweep controller of magnetic field, display, etc. In the present preliminary study, reproducibility values are measured with the same ESR parameters and alanine-polystyrene (alanine-PS) dosimeter at a dose of 1 kGy: repeatedly measuring without removing dosimeter from the cavity; individual measurement with removing and inserting again into the cavity with readjustment of ESR parameters. Alanine/ESR dosimetry using this spectrometer has a measurable dose range from 1 to 100 kGy with relatively high precision within ± 3% (1σ) as a preliminary result. The portable ESR spectrometer may also be modified as an automatic, more precise, dedicated alanine dosimeter reader. (author)

  11. The relationship of alanine aminotransferase to metabolic syndrome in a Korean population.

    Science.gov (United States)

    Lee, Seok Hoon; Cho, Doo Yeoun; Joo, Nam Seok; Kim, Kwang Min; Kim, Kyu Nam

    2018-01-01

    Although associations between serum alanine aminotransferase and metabolic syndrome are well-recognized in Western countries, only a limited number of prospective studies have been performed in Asian populations. The aim of the study was to cross-sectionally and longitudinally examine whether serum alanine aminotransferase levels are associated with metabolic syndrome and its associated components in a Korean population. A total of 31,832 subjects who received health screenings were included in cross-sectional analyses; a subgroup of 4.070 subjects without metabolic syndrome at baseline was included in the longitudinal analyses. The metabolic syndrome definition was based on the National Cholesterol Education Program Third Adult Treatment Panel criteria with modification on waist circumference cut-off to be more appropriate for an Asian population. In the cross-sectional analyses, serum alanine aminotransferase is positively associated with metabolic syndrome and its components. In the longitudinal analyses, the prevalence of metabolic syndrome increased across serum alanine aminotransferase quartiles in a dose-dependent manner after extensive adjustments (hazard ratios were 1.000, 1.609, 2.601, and 3.015 for quartiles, 1 through quartile 4; P for trendmetabolic syndrome and elevated serum alanine aminotransferase in a Korean population.

  12. 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

    OpenAIRE

    Kim, Seryoung; Ihara, Kohei; Katsube, Satoshi; Hori, Hatsuhiro; Ando, Tasuke; Isogai, Emiko; Yoneyama, Hiroshi

    2015-01-01

    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 intr...

  13. 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

  14. Mechanism of mercurial inhibition of sodium-coupled alanine uptake in liver plasma membrane vesicles from Raja erinacea

    International Nuclear Information System (INIS)

    Sellinger, M.; Ballatori, N.; Boyer, J.L.

    1991-01-01

    In mammalian hepatocytes the L-alanine carrier contains a sulfhydryl group that is essential for its activity and is inhibited by mercurials. In hepatocytes of the evolutionarily primitive little skate (Raja erinacea), HgCl2 inhibits Na(+)-dependent alanine uptake and Na+/K(+)-ATPase and increase K+ permeability. To distinguish between direct effects of HgCl2 on the Na(+)-alanine cotransporter and indirect effects on membrane permeability, [3H]alanine transport was studied in plasma membrane vesicles. [3H]Alanine uptake was stimulated by an out-to-in Na+ but not K+ gradient and was saturable confirming the presence of Na(+)-alanine cotransport in liver plasma membranes from this species. Preincubation of the vesicles with HgCl2 for 5 min reduced initial rates of Na(+)-dependent but not Na(+)-independent alanine uptake in a dose-dependent manner (10-200 microM). In the presence of equal concentrations of NaCl or KCl inside and outside of the vesicles, 75 microM HgCl2 directly inhibited sodium-dependent alanine-[3H]alanine exchange, demonstrating that HgCl2 directly affected the alanine cotransporter. Inhibition of Na(+)-dependent alanine uptake by 30 microM HgCl2 was reversed by dithiothreitol (1 mM). HgCl2 (10-30 microM) also increased initial rates of 22Na uptake (at 5 sec), whereas 22Na uptake rates were decreased at HgCl2 concentrations greater than 50 microM. Higher concentrations of HgCl2 (100-200 microM) produced nonspecific effects on vesicle integrity. These studies indicate that HgCl2 inhibits Na(+)-dependent alanine uptake in skate hepatocytes by three different concentration-dependent mechanisms: direct interaction with the transporters, dissipation of the driving force (Na+ gradient), and loss of membrane integrity

  15. Stability constants of mixed ligand complexes of dioxouranium(II) and thorium(IV) with complexones and isomeric alanines

    International Nuclear Information System (INIS)

    Singh, R.K.; Saxena, M.C.

    1992-01-01

    The present work reports on the stability sequence between UO 2 II and Th IV ions for their mixed ligands complexes with the two isomeric alanines, α-alanine (α-ala) and β-alanine (β-ala) containing a complexone as primary ligand. The complexones used are iminodiacetate (IMDA), nitrilotricetate (NTA), 2-hydroxyethylenediaminetriacetate (HEDTA), ethylenediaminetetraacetate (EDTA), 1,2-diaminocyclohexanetraacetate (CDTA) and diethylenetriminepentaacetate (DTPA). (author). 9 refs., 1 tab

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

    International Nuclear Information System (INIS)

    Ketata, Ameni

    2011-01-01

    The electron spin resonance spectroscopy is well known as a reference and transfer dosimetry system for the evaluation of high doses in radiation processing. The high cost of an EPR/alanine dosimetry system is a serious handicap for large-scale routine application. In this study, the use of irradiated L-alanine dissolved in color indicator solutions (bromothymol blue and fuchsin) was investigated for dosimetry purposes. This solution has an absorbance varies linearly with the absorbed dose in the dose range of 0-25 kGy for the bromothymol blue, and 0-45 kGy for the fuchsin. The effects of the dye and the alanine concentration, the p H value as well as of the solvent have been studied. With respect to routine application, the stability of dosimeters was also investigated

  17. Degradation of pyrimidines in Saccharomyces kluyveri: transamination of beta-alanine

    DEFF Research Database (Denmark)

    Schnackerz, K D; Andersen, G; Dobritzsch, D

    2008-01-01

    Beta-alanine is an intermediate in the reductive degradation of uracil. Recently we have identified and characterized the Saccharomyces kluyveri PYD4 gene and the corresponding enzyme beta -alanine aminotransferase ((Sk)Pyd4p), highly homologous to eukaryotic gamma-aminobutyrate aminotransferase...... (GABA-AT). S. kluyveri has two aminotransferases, GABA aminotransferase ((Sk)Uga1p) with 80% and (Sk)Pyd4p with 55% identity to S. cerevisiae GABA-AT. (Sk)Pyd4p is a typical pyridoxal phosphate-dependent aminotransferase, specific for alpha-ketoglutarate (alpha KG), beta-alanine (BAL) and gamma......-aminobutyrate (GABA), showing a ping-pong kinetic mechanism involving two half-reactions and substrate inhibition. (Sk)Uga1p accepts only alpha KG and GABA but not BAL, thus only (Sk)Pydy4p belongs to the uracil degradative pathway....

  18. Densities and solubilities of Glycylglycine and Glycyl-L-Alanine in Aqueous Electrolyte Solutions

    DEFF Research Database (Denmark)

    Breil, Martin Peter; Mollerup, Jørgen; Rudolph, E. Susanne J.

    2004-01-01

    Solubilities of glycylglycine and glycyl-L-alanine in aqueous electrolyte solutions containing 0-6 molal NaCl, 0-1 molal Na2SO4, and 0-1 molal (NH4)(2)SO4, have been determined experimentally at 298.15 K and atmospheric pressure. The solubility of glycylglycine and glycyl-L-alanine in pure water...... is 1.74 and 4.78 mol/kg of water, respectively. The solubility of glycylglycine in salt solutions of NaCl, Na2SO4, and (NH4)(2)SO4 show a moderate salting-in effect. The solubility of glycyl-L-alanine show a minor or no salting-in effect at low salt concentrations and a moderate salting-out effect...

  19. 14N nuclear quadrupole interaction in Cu(II) doped L-alanine

    International Nuclear Information System (INIS)

    Murgich, J.; Calvo, R.; Oseroff, S.B.; Instituto Venezolano de Investigaciones Cientificas, Caracas. Dept. de Quimica)

    1980-01-01

    The 14 N nuclear quadrupole interaction tensor Psub(N) measured by ENDOR in Cu(II) doped L-alanine is analyzed in terms of the Townes and Daily theory assuming a tetra-hedrally bonded N atom. The results of this analysis are compared with those for the 14 N in pure L-alanine and it is found that the principal directions of the Psub(N) tensor are drastically changed upon metal complexation as a consequence of the higher electron affinity of Cu(II) with respect to C and H. Comparison of the corresponding bond populations in pure and Cu(II) doped L-alanine indicates that the Cu draws 0.11 more electron from the N than the substituted H atom. (orig.)

  20. Diastereoselective discrimination of lysine-alanine-alanine peptides by zwitterionic cinchona alkaloid-based chiral selectors using electrospray ionization mass spectrometry.

    Science.gov (United States)

    Bobbitt, Jonathan M; Li, Li; Carlton, Doug D; Yasin, Mahwish; Bhawal, Sumit; Foss, Frank W; Wernisch, Stefanie; Pell, Reinhard; Lindner, Wolfgang; Schug, Kevin A

    2012-12-21

    Electrospray ionization-mass spectrometry (ESI-MS) was used to investigate stereoselective interactions between seven zwitterionic alkylsulfonate-modified cinchona alkaloid chiral selectors and biologically relevant lysine-alanine-alanine tripeptide and alanine-alanine dipeptide selectands in modified methanolic solutions. Ion intensities from full scan mass spectra were used to assess degrees of association, the ratios of which were used to calculate selectivities for different selector-selectand pairs. The results support prior work on similar systems using HPLC, in that binding is mediated in these systems primarily through the quinuclidine amine on the selector and the C-terminal carboxylate of the peptide. N(α)- and N(α), N(ɛ)-acetylated forms of the tripeptide were used to study the relative contribution to binding imparted by the presence of multiple basic amines on the tripeptide with the selectors; this was not previously investigated by HPLC. The ability of the sulfonate group on the selector to reach and preferentially interact with the N(ɛ)-amine on the side chain of lysine was revealed. Overall, in acidic methanol conditions (0.5% acetic acid), degrees of association ranged from 1.5% to 17%, and selectivities ranged from non-selective to a 5.5:1 preference for binding one peptide stereoisomer over another with a given chiral selector. In sodium acetate (100 μM)-modified methanol solutions, significant changes in degrees of association (ranging from 4% to 25%) and selectivities (ranging from non-selective to 4.2:1 preference) were observed. These mass spectrometry experiments help to clarify the chiral recognition mechanism for these selectors and suggest that retention and selectivity could be further modulated in HPLC experiments through the utilization of alkali salt-containing mobile phases. Copyright © 2012 Elsevier B.V. All rights reserved.

  1. Relative response of the alanine dosimeter to medium energy x-rays.

    Science.gov (United States)

    Anton, M; Büermann, L

    2015-08-07

    The response of the alanine dosimeter to kilovoltage x-rays with respect to the dose to water was measured, relative to the response to Co-60 radiation.Two series of x-ray qualities were investigated, one ranging from 30 kV to 100 kV tube voltage (TW series), the other one ranging from 70 kV to 280 kV (TH series). Due to the use of the water calorimeter as a primary standard, the uncertainty of the delivered dose is significantly lower than for other published data. The alanine response was measured as described in a previous publication (Anton et al 2013 Phys. Med. Biol. 58 3259-82). The uncertainty component due to the alanine measurement and analysis is ⩽0.4%, the major part of the combined uncertainty of the relative response originates from the uncertainty of the delivered dose. The relative uncertainties of the relative response vary from ⩽2% for the TW series to ⩽1.1% for the TH series.Different from the behaviour of the alanine dosimeter for megavoltage x-rays or electrons, the relative response drops significantly from unity for Co-60 radiation to less than 64% for the TW quality with a tube voltage of 30 kV. In order to reproduce this behaviour through Monte Carlo simulations, not only the ratio of the absorbed dose to alanine to the absorbed dose to water has to be known, but also the intrinsic efficiency, i.e. the dependence of the number of free radicals generated per unit of absorbed dose on the photon energy. This quantity is not yet accessible for the TW series.For a possible use of the alanine dosimeter for kilovoltage x-rays, for example in electronic brachytherapy, users should rely on the measured data for the relative response which have become available with this publication.

  2. 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 +

  3. End-to-end tests using alanine dosimetry in scanned proton beams

    Science.gov (United States)

    Carlino, A.; Gouldstone, C.; Kragl, G.; Traneus, E.; Marrale, M.; Vatnitsky, S.; Stock, M.; Palmans, H.

    2018-03-01

    This paper describes end-to-end test procedures as the last fundamental step of medical commissioning before starting clinical operation of the MedAustron synchrotron-based pencil beam scanning (PBS) therapy facility with protons. One in-house homogeneous phantom and two anthropomorphic heterogeneous (head and pelvis) phantoms were used for end-to-end tests at MedAustron. The phantoms were equipped with alanine detectors, radiochromic films and ionization chambers. The correction for the ‘quenching’ effect of alanine pellets was implemented in the Monte Carlo platform of the evaluation version of RayStation TPS. During the end-to-end tests, the phantoms were transferred through the workflow like real patients to simulate the entire clinical workflow: immobilization, imaging, treatment planning and dose delivery. Different clinical scenarios of increasing complexity were simulated: delivery of a single beam, two oblique beams without and with range shifter. In addition to the dose comparison in the plastic phantoms the dose obtained from alanine pellet readings was compared with the dose determined with the Farmer ionization chamber in water. A consistent systematic deviation of about 2% was found between alanine dosimetry and the ionization chamber dosimetry in water and plastic materials. Acceptable agreement of planned and delivered doses was observed together with consistent and reproducible results of the end-to-end testing performed with different dosimetric techniques (alanine detectors, ionization chambers and EBT3 radiochromic films). The results confirmed the adequate implementation and integration of the new PBS technology at MedAustron. This work demonstrates that alanine pellets are suitable detectors for end-to-end tests in proton beam therapy and the developed procedures with customized anthropomorphic phantoms can be used to support implementation of PBS technology in clinical practice.

  4. Synthesis and characterization of alanine boron hydrate for its use in thermal neutron dosimetry

    International Nuclear Information System (INIS)

    Yanez S, J.C.

    1994-01-01

    Alanine boron hydrate was synthesized for its possible use as intercomparison dosimeter for thermal neutron irradiation. The irradiations were performed in the Nuclear Reactor of the Nuclear Center of Mexico. The salt was prepared by reacting alanine and boric acid in a (1:1) stoichiometric ratio in neutral pH 7.5 aqueous solution and also in a basic pH 13 solution. The latter reaction was prepared with the addition of ammonia hydroxide (25%). Solutions were stirred and afterwards were let to evaporate. The obtained product in each reaction is a white solid. Dosimeters were prepared with the obtained reaction products and irradiated under thermal neutron flux of 5 x 10 7 n/cm 2 s. For 30 hours. The analysis of irradiated samples was made in a Variant E-15 Electron Paramagnetic Resonance spectrometer. The observed response of the samples prepared with the reaction product at the basic pH is approximately 50% higher than the neutral pH samples. In order to investigate the optimum signal enhancement samples were prepared in a basic pH medium in the following stoichiometric ratios: (1:0.5); (1:0.75); (1:1.25); (1:1.5) and (1:1.75). It was observed that the samples of the reaction (1:0.75) produced the higher response. The response was 2728% higher than the alanine only dosimeters. The reaction product was chemically characterized by X-ray diffraction, Nuclear Magnetic Resonance, Chromatography, Refractometry and Solubility tests. Results indicate that alanine boron hydrate is formed in basic media and in a stoichiometric ratio (1:0.75). The dosimetric characterization of alanine boron hydrate was performed, results are reported. It is concluded that alanine boron hydrate may be a good intercomparison dosimeter for thermal neutron irradiation. (Author)

  5. 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)

  6. A spectrophotometric readout for γ irradiated alanine solution - a dosimetric application

    International Nuclear Information System (INIS)

    Marzouk, Asma

    2007-01-01

    Alanine is a stable dosimeter of reference in its solid state. Its installation in solution as being a dosimetric system of routine remains very useful. A follow-up of the behaviour of the irradiated alanine solution with 15 kGy according to the concentration is carried out by UV-Visible spectrophotometry. The results obtained prove the difficulty in analytical studies of the radiolysis of aqueous solutions by optical absorption due to the ambiguous broad spectra of the species and the reaction products. (Author). 47 refs

  7. On the fragmentation of biomolecules: Fragmentation of alanine dipeptide along the polypeptide chain

    International Nuclear Information System (INIS)

    Solov'yov, I. A.; Yakubovich, A. V.; Solov'yov, A. V.; Greiner, W.

    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 fragmentation of dipeptide along the polypeptide chain, as well as the interaction between alanines, has been considered. The energy of the system has been analyzed as a function of the distance between fragments for all possible dipeptide fragmentation channels. Analysis of the energy barriers makes it possible to estimate the characteristic fragmentation times and to determine the degree of applicability of classical electrodynamics for describing the system energy

  8. No effect of β-alanine on muscle function and kayak performance

    DEFF Research Database (Denmark)

    Bech, Signe Refsgaard; Nielsen, Tobias Schmidt; Hald, Martin

    2018-01-01

    PURPOSE: It was investigated if β-alanine supplementation counteracts muscular fatigue development or improves athletic performance. METHODS: Elite kayak rowers (10 males and 7 females) were supplemented with either 80 mg/kg body mass/day β-alanine or placebo for 8 weeks. Muscular fatigue...... development was investigated by applying a 2 min elbow flexor maximal voluntary contraction (MVC). Electromyography (EMG) was recorded continuously and voluntary activation (VA) was determined 30, 60, 90 and 115 s into the 2 min MVC. Additionally, performance was evaluated as 1000 m and 5 × 250 m kayak...

  9. The photon energy dependence of the alanine/EPR dosimetry system, an experimental investigation

    International Nuclear Information System (INIS)

    Bergstrand, E.S.; Hole, E.O.; Shortt, K.R.; Ross, C.K.

    2002-01-01

    The energy dependence of a dosimetry system based on electron paramagnetic resonance (EPR) spectroscopy of alanine has been studied to determine its suitability for use in dose verification for radiotherapy. A few experiments with high-energy photon irradiation of alanine have been reported in the literature. However, the reported results disagree whether the ratio of dose in alanine to dose in water is independent of the radiation energy or whether there is a small dependence for photon energies of relevance to radiotherapy. The concentration of free radicals in alanine is proportional to the absorbed dose in alanine over a wide dose range covering three decades. The relative number of radicals may be determined by examining the EPR spectrum, and hence it is possible to determine the dose with a system that has been calibrated using a known dose of 60 Co radiation. In the present work, irradiations of alanine dosimeters were performed at the National Research Council (NRC), in Ottawa, Canada. The radiation qualities investigated were 10, 20 and 30 MV x-rays using the NRC linac. For each radiation quality, 30 dosimeters were irradiated in a water phantom with a level of absorbed dose to water ranging from 10 to 50 Gy. For reference purposes, irradiations using the NRC 60 Co source were performed on more or less the same day as the irradiations at each specific linac quality. In all beams, the dose to water was measured using a graphite-walled NE2571 ionisation chamber that was originally calibrated by comparison with a sealed-water calorimeter. The alanine dosimeters were evaluated at the EPR laboratory at the University of Oslo, Norway, using an X-band Bruker ESP300E spectrometer with a rectangular double resonator. One of the resonators contained a Mn 2+ /MgO sample that was read after each dosimeter reading, in order to provide independence from short-term sensitivity fluctuations in the spectrometer. All dosimeters irradiated at one specific linac quality were

  10. An investigation of the photon energy dependence of the EPR alanine dosimetry system

    International Nuclear Information System (INIS)

    Bergstrand, Eva Stabell; Shortt, Ken R; Ross, Carl K; Hole, Eli Olaug

    2003-01-01

    The electron paramagnetic resonance (EPR) alanine dosimetry system is based on EPR measurements of radicals formed in alanine by ionizing radiation. The system has been studied to determine its energy dependence for photons in the 10-30 MV region relative to those of 60 Co and to find out if the system would be suitable for dosimetry comparisons. The irradiations were carried out at the National Research Council, Ottawa, Canada and the doses ranged from 8 to 54 Gy. The EPR measurements were performed at the University of Oslo, Norway. The ratio of the slope of the alanine reading versus dose-to-water curve for a certain linac photon beam quality and the corresponding slope for a reference 60 Co γ-radiation gives an experimental measure of the relative dose-to-water response of the EPR alanine dosimetry system. For calculating the linear regression coefficients of these alanine reading versus dose curves, the method of weighted least squares was used. This method is assumed to produce more accurate regression coefficients when applied to EPR dosimetry than the common method of standard least squares. The overall uncertainty on the ratio of slopes was between 0.5 and 0.6% for all three linac energies. The relative response for all the linac beams compared to cobalt was less than unity: by about 0.5% for the 20 and 30 MV points but by more than 1% for the 10 MV point. The given standard uncertainties negate concluding that there is any significant internal variation in the measured response as a function of beam quality between the three linac energies. Thus, we calculated the average dose response for all three energies and found that the alanine response is 0.8% (±0.5%) lower for high energy x-rays than for 60 Co γ-rays. This result indicates a small energy dependence in the alanine response for the high-energy photons relative to 60 Co which may be significant. This result is specific to our dosimetry system (alanine with 20% polyethylene binder pressed into a

  11. Development of a dosimeter for high doses assessment based on Alanine/EPR

    International Nuclear Information System (INIS)

    Galante, O.L.; Rodrigues, O. Jr.; Campos, L.L.

    2000-01-01

    The increasing use of radiation sources of high activity for industrial and medical applications becomes important the research and the development of detectors and dosimetric methods for quality control of the applied doses. This work presents the current stage of the research at IPEN/CNEN-SP that has as objective the development of a standard dosimetric system for high doses assessment based on the alanine as radiation detector and electron paramagnetic resonance (EPR) as measurement technique. The developed system consists of the cylindrical container built in polyethylene of high density and the detector element based on DL-alanine commercially available. For the detector preparation different binding materials such as paraffin and acetate polyvinyl solution (pva) and also the use of a polyethylene tube of low density with 3.2 mm of external diameter, 2 mm of internal diameter and 30 mm of length were tested to provide the easier preparation method and the most sensitive detector. For the alanine + paraffin detector it was used 80% of alanine and 20% of paraffin, for the alanine + pva detector it was used 70% of alanine and 30% of pva solution, and pure alanine was encapsulated, compacted and sealed in the case of the polyethylene tube. The obtained results with respect to handling, packing and construction easiness showed that the polyethylene tube presents all characteristics to obtain of a good detector element. The validation of the dosimetric system was carried out with gamma radiation of the cobalt-60 with doses in the range between 0.2 Gy to 200 kGy. Type tests such as fading, lowest detection limit, reproducibility and energy dependence of the sign EPR were performed. All measurements were carried out at room temperature using a spectrometer of electron paramagnetic resonance (EPR) Bruker model MXE. Taking into account the results obtained: linearity of the EPR signal between 10 Gy and 50 kGy, reproducibility better than 3%, low fading associated with

  12. Effect of combined β-alanine and sodium bicarbonate supplementation on cycling performance.

    Science.gov (United States)

    Bellinger, Phillip M; Howe, Samuel T; Shing, Cecilia M; Fell, James W

    2012-08-01

    The purpose of this study was to investigate the effects of 28 d of β-alanine supplementation on 4-min cycling time trial performance and to determine whether there was an additive effect of combined β-alanine and sodium bicarbonate (NaHCO3) supplementation on high-intensity cycling performance. Fourteen highly trained cyclists (mean ± SD: age = 25.4 ± 7.2 yr, mass = 71.1 ± 7.1 kg, V˙O(2max) = 66.6 ± 5.7 mL·kg·min) supplemented for 28 d with β-alanine (65 mg·kg body mass each day) or placebo. A maximal 4-min bout of cycling was performed before supplementation (baseline) and twice after supplementation: after ingestion of NaHCO3 (300 mg·kg body mass) and ingestion of a placebo using a randomized crossover design with 2 d between trials. Blood pH and HCO3 concentration were determined before loading (postsupplementation trials) and at pretest and posttest. In the acute NaHCO3 loading trials, blood pH and HCO3 were elevated from before loading to pretest, and the magnitude of the change in HCO3 from pretest to posttest was significantly greater compared with the acute placebo loading trial (P < 0.001). Average power output in the 4-min cycling performance trial was increased in placebo + NaHCO3 (+3.1% ± 1.8%) and β-alanine + NaHCO3 (+3.3% ± 3.0%) compared with baseline (P < 0.05). β-alanine + placebo did not significantly improve average power output compared with baseline (+1.6% ± 1.7%, P = 0.20); however, magnitude-based inferences demonstrated that β-alanine + placebo was associated with a 37% likelihood of producing average power improvements. In trained cyclists, β-alanine supplementation did not significantly improve 4-min cycling performance; however, there may be a small meaningful improvement in performance. Acute NaHCO3 supplementation significantly improved 4-min cycling performance. There seemed to be a minimal additive effect of combined β-alanine and NaHCO3 supplementation.

  13. Synthesis, Characterization and Metal Ion Detection of Novel Fluoroionophores Based on Heterocyclic Substituted Alanines

    Directory of Open Access Journals (Sweden)

    M. Manuela M Raposo

    2007-10-01

    Full Text Available The synthesis of new fluorescent probes containing the thiophene andbenzoxazole moieties combined with an alanine residue is described. The resulting highlyfluorescent heterocyclic alanine derivatives respond via a quenching effect, withparamagnetic Cu(II and Ni(II metal ions and with diamagnetic Hg(II, as shown by theabsorption and steady-state fluorescence spectroscopy studies. The formation ofmononuclear or dinuclear metal complexes was postulated based on the presence of thefree carboxylic acid as binding site and also with the interaction with the donor atoms inthe chromophore. Interaction with other important biological metal ions such as Zn(II,Ca(II and Na(I was also explored.

  14. Nationwide survey to compare the prevalence of transient elevation of liver transaminase during treatment of diabetic ketosis or ketoacidosis in new-onset acute and fulminant type 1 diabetes mellitus.

    Science.gov (United States)

    Takaike, Hiroko; Uchigata, Yasuko; Iwamoto, Yasuhiko; Imagawa, Akihisa; Iwahashi, Hiromi; Kanatsuka, Azuma; Kawasaki, Eiji; Kobayashi, Tetsuro; Shimada, Akira; Shimizu, Ikki; Maruyama, Taro; Hanafusa, Toshiaki; Makino, Hideichi

    2008-01-01

    A mild increase in liver enzyme levels is sometimes observed in patients with diabetic ketosis or ketoacidosis. The aim of the present study was to assess the cause and prevalence of the elevation of liver transaminase levels in fulminant and acute-onset type 1 diabetic patients experiencing diabetic ketosis or ketoacidosis. We analyzed data on the liver transaminase levels of 108 patients over 18 years of age with newly diagnosed type 1 diabetes complicated by ketosis or ketoacidosis. The data were collated from a nationwide survey on fulminant type 1 diabetes and retrospective medical records. Thirty-two (60.4%) out of the 53 patients suffering from fulminant type 1 diabetes were detected with transient elevation of liver transaminase (TELT) levels during the first month after initiation of insulin therapy; in the case of acute-onset type 1 diabetes, such an observation was noted in 16 (29.1%) out of 55 patients. Fatty liver was diagnosed in 20% of the patients, and 65% of these patients exhibited TELT. The dosage of insulin injected in these patients was significantly high. High blood glucose and fatty liver may influence the elevation of liver transaminase levels during the treatment of new-onset type 1 diabetes.

  15. Advancements in accuracy of the alanine dosimetry system. Part 1. The effects of environmental humidity

    International Nuclear Information System (INIS)

    Sleptchonok, Olga F.; Nagy, Vitaly; Desrosiers, Marc F.

    2000-01-01

    A one-year study of the EPR signal of γ-irradiated ( 60 Co) L-α-alanine with simultaneous monitoring of the cavity Q-factor was undertaken. The widespread opinion that the EPR signal remains absolutely stable under normal laboratory storage conditions is inaccurate. At 0% humidity, the signal can be regarded as stable within ±1% of its initial value for 6 months for 1 and 10 kGy doses, but for only 3 months for 100 kGy. When stored at the same relative humidity values up to 60%, the fading rates for dosimeters irradiated to 1 and 10 kGy are similar, whereas signals of dosimeters irradiated to 100 kGy fade considerably faster for all humidities. The rates of fading increase with the relative humidity, especially above 60% R. H. Environmental humidity also deteriorates the accuracy of alanine dosimetry by changing the resonant cavity Q-factor. This is particularly important when irradiated alanine dosimeters are used as instrument calibration standards. Short-term changes in alanine EPR signal amplitudes were recorded upon removal of the irradiated dosimeters from their storage environments. The importance of an in situ standard to correct for measurement errors due to environmental effects is demonstrated. (author)

  16. Inhibitory effect of different product analogues on β-alanine synthase: A thermodynamic and fluorescence analysis

    International Nuclear Information System (INIS)

    Andujar-Sanchez, Montserrat; Martinez-Gomez, Ana Isabel; Martinez-Rodriguez, Sergio; Clemente-Jimenez, Josefa Maria; Heras-Vazquez, Francisco Javier Las; Rodriguez-Vico, Felipe; Jara-Perez, Vicente

    2009-01-01

    The enzyme N-carbamoyl-β-alanine amidohydrolase catalyse the hydrolysis of N-carbamoyl-β-alanine or N-carbamoyl-β-aminoisobutyric acid to β-alanine or 3-aminoisobutyric acid, under the release of carbon-dioxide and ammonia. This work studies the inhibition of N-carbamoyl-β-alanine amidohydrolase from Agrobacterium tumefaciens C58 (Atβcar) by different carboxylic acid compounds that differ in number of carbons, and position and size of ramification, while the binding thermodynamics of the inhibitors are studied by isothermal titration calorimetry (ITC) and fluorescence. From the binding constants and inhibition studies, we conclude that propionate is the most efficient inhibitor among those tested. Substitution of the linear alkyl acids in positions 2 and 3 resulted in a drastic decrease of the affinity. The thermodynamic parameters show that a conformational change is triggered upon ligand binding. Binding enthalpy ΔH b is negative in all cases for all ligands, and thus, Van der Waals interactions and hydrogen bonding are most probably the major sources for this term. The process is entropically favoured at all temperatures and pH studied, most probably due to the liberation of water molecules accompanying the conformational change of the enzyme

  17. High-pressure X-ray diffraction of L-ALANINE crystal

    DEFF Research Database (Denmark)

    Olsen, J.S.; Gerward, Leif; Souza, A.G.

    2006-01-01

    L-ALANINE has been studied by X-ray diffraction at ambient temperature and pressure up to 10.3 GPa. The material is found to transform to a tetragonal structure between 2 and 3 GPa. and to a monoclinic structure between 8 and 10 GPa. The experimental bulk modulus is 25(5) GPa for the orthorhombic...

  18. Inhibitory effect of different product analogues on {beta}-alanine synthase: A thermodynamic and fluorescence analysis

    Energy Technology Data Exchange (ETDEWEB)

    Andujar-Sanchez, Montserrat; Martinez-Gomez, Ana Isabel; Martinez-Rodriguez, Sergio; Clemente-Jimenez, Josefa Maria; Heras-Vazquez, Francisco Javier Las; Rodriguez-Vico, Felipe [Departamento de Quimica Fisica, Bioquimica y Quimica Inorganica, Facultad de Ciencias Experimentales, Universidad de Almeria, Carretera de Sacramento s/n, La Canada de San Urbano, Almeria 04120 (Spain); Jara-Perez, Vicente [Departamento de Quimica Fisica, Bioquimica y Quimica Inorganica, Facultad de Ciencias Experimentales, Universidad de Almeria, Carretera de Sacramento s/n, La Canada de San Urbano, Almeria 04120 (Spain)], E-mail: vjara@ual.es

    2009-02-15

    The enzyme N-carbamoyl-{beta}-alanine amidohydrolase catalyse the hydrolysis of N-carbamoyl-{beta}-alanine or N-carbamoyl-{beta}-aminoisobutyric acid to {beta}-alanine or 3-aminoisobutyric acid, under the release of carbon-dioxide and ammonia. This work studies the inhibition of N-carbamoyl-{beta}-alanine amidohydrolase from Agrobacterium tumefaciens C58 (At{beta}car) by different carboxylic acid compounds that differ in number of carbons, and position and size of ramification, while the binding thermodynamics of the inhibitors are studied by isothermal titration calorimetry (ITC) and fluorescence. From the binding constants and inhibition studies, we conclude that propionate is the most efficient inhibitor among those tested. Substitution of the linear alkyl acids in positions 2 and 3 resulted in a drastic decrease of the affinity. The thermodynamic parameters show that a conformational change is triggered upon ligand binding. Binding enthalpy {delta}H{sub b} is negative in all cases for all ligands, and thus, Van der Waals interactions and hydrogen bonding are most probably the major sources for this term. The process is entropically favoured at all temperatures and pH studied, most probably due to the liberation of water molecules accompanying the conformational change of the enzyme.

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

    Directory of Open Access Journals (Sweden)

    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.

  20. 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

  1. 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)

  2. Influence of the irradiation temperature on the free-radical response of alanine

    International Nuclear Information System (INIS)

    Wieser, A.; Siegele, R.; Regulla, D.F.

    1989-01-01

    GSF operates the only IAEA high-level dosimetry reference laboratory and, as a joint project with the IAEA, the International Dose Assurance Service (IDAS). Dosimetry is based on long-lived free radicals in organic alanine induced by ionizing radiation and readout by ESR spectroscopy. The thermal time response of the radical concentration in alanine is fairly constant after irradiation provided that the alanine samples are stored at temperatures below 50 0 C. By contrast, a positive temperature coefficient had earlier been found at GSF for the production rate of alanine radicals, for irradiation temperatures between 0 and 50 0 C. This effect has to be considered for reference dosimetry in radiation processing. Radiation processing is also of interest at irradiation temperatures below 0 0 C. The present study describes experiments on the influence of irradiation temperatures between +50 and -100 0 C. Comparison is made between the present and earlier results, in the overlapping temperature range. An empirical function is proposed for the temperature coefficient based on the experimental data. (author)

  3. 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

    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...

  4. Spectrophotometric readout for alanine dosimeter to be used for food irradiation applications

    International Nuclear Information System (INIS)

    Ebraheem, S.; Beshir, W.B.; Sobhy, R.; Kovacs, A.; Wojnarovits, L.

    2002-01-01

    Complete text of publication follows. The electron spin resonance (EPR) readout of radical concentration produced upon irradiation of L-alanine is well known as a transfer dosimetry system for high dose range. The high cost of EPR/alanine dosimetry system is a serious handicap for large-scale routine application for use in irradiation facilities. In this study the irradiated L-alanine was dissolved in 1,4 -phenyl diammonium dichloride (PAC) solution resulting a complex of a purple colour. This complex has a variable absorbance with applied dose in the range from 1 - 20 kGy. The absorbance of this complex using spectrophotometric evaluation was investigated at 401 and 478 nm with increasing dose. The fluorimetric emission was also studied at the wavelength of 475 nm as a function of dose. This methos for dose evaluation is uncomplicated and very easy for routine application. The effect of the dye concentration as well as the suitable amount of alanine has been studied with respect to practical use. The stability of the complex for long period of time was also investigated

  5. Effect of chronic renal failure with metabolic acidosis on alanine metabolism in isolated liver cells

    NARCIS (Netherlands)

    Cano, N.; Sturm, J. M.; Meijer, A. J.; El-Mir, M. Y.; Novaretti, R.; Reynier, J. P.; Leverve, X. M.

    2004-01-01

    Background Et aims: Decreased ureagenesis and gluconeogenesis from atanine have been reported during chronic renal failure in rat. This study addressed the respective roles of plasma-membrane transport and intracellular metabolism in these abnormalities of alanine pathways. Methods: In hepatocytes

  6. 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.

  7. Beta-alanine/alpha-ketoglutarate aminotransferase for 3-hydroxypropionic acid production

    Science.gov (United States)

    Jessen, Holly Jean [Chanhassen, MN; Liao, Hans H [Eden Prairie, MN; Gort, Steven John [Apple Valley, MN; Selifonova, Olga V [Plymouth, MN

    2011-10-04

    The present disclosure provides novel beta-alanine/alpha ketoglutarate aminotransferase nucleic acid and protein sequences having increased biological activity. Also provided are cells containing such enzymes, as well as methods of their use, for example to produce malonyl semialdehyde and downstream products thereof, such as 3-hydroxypropionic acid and derivatives thereof.

  8. The VA and VCD spectra of various isotopomers of L-alanine in aqueous solution

    DEFF Research Database (Denmark)

    Abdali, Salim; Jalkanen, Karl J.; Bohr, Henrik

    2002-01-01

    Density functional theory (DFT) at the Becke 3LYP level has been used to calculate the vibrational absorption (VA) and vibrational circular dichroism (VCD) spectra of various deuterated species Of L-alanine. The effect of replacing the methine hydrogen, CH1, the methyl group, CH3, and both...

  9. Synthesis, characterization, and biocompatible properties of alanine-grafted chitosan copolymers.

    Science.gov (United States)

    Park, Gyu Han; Kang, Min-Sil; Knowles, Jonathan C; Gong, Myoung-Seon

    2016-04-01

    In order to overcome major problems regarding the lack of affinity to solvents and limited reactivity of the free amines of chitosan, introduction of appropriate spacer arms having terminal amine function is considered of interest. L-Alanine-N-carboxyanhydride was grafted onto chitosan via anionic ring-opening polymerization. The chemical and structural characterizations of L-alanine-grafted chitosan (Ala-g-Cts) were confirmed through Fourier transform infrared spectroscopy and proton nuclear magnetic resonance spectroscopy ((1)H NMR). In addition, the viscoelastic properties of Ala-g-Cts were examined by means of a rotational viscometer, and thermal analysis was carried out with a thermogravimetric analyzer and differential scanning calorimetry. Morphological changes in the chitosan L-alanine moiety were determined by x-ray diffraction. To determine the feasibility of using these films as biomedical materials, we investigated the effects of their L-alanine content on physical and mechanical properties. The biodegradation results of crosslinked Ala-g-Cts films were evaluated in phosphate-buffered solution containing lysozyme at 37℃. Proliferation of MC3T3-E1 cells on crosslinked Ala-g-Cts films was also investigated with use of the CCK-8 assay. © The Author(s) 2016.

  10. Effect of alpha interferon on glucose and alanine transport by rat renal brush border membrane vesicles

    International Nuclear Information System (INIS)

    Batuman, V.; Chadha, I.

    1990-01-01

    To investigate the pathogenetic mechanisms of interferon nephrotoxicity, we studied the effect of recombinant interferon alfa-2b on the uptake of 14 C-D-glucose and 14 C-L-alanine by rat renal brush-border-membrane vesicles. Interferon significantly inhibited 20 sec. sodium-dependent and 5 and 10 min. equilibrium uptake of both glucose and alanine. The inhibitory effect was dose dependent with maximum effect achieved at interferon concentration of 5 x 10 -8 M in the uptake media. The half-maximal inhibitory concentrations, IC 50 , of interferon on glucose uptake was 1.8 x 10 -8 M, and 5.4 x 10 -9 M on alanine uptake. Dixon plot analysis of uptake data was consistent with pure non-competitive inhibition. The inhibition constants, K i , 1.5 x 10 -8 M for glucose uptake, and 7.3 x 10 -9 M for alanine uptake, derived from Dixon plots were in close agreement with the IC 50 s calculated from the semilog dose response curves. These observations reveal that direct interactions at the proximal tubule cell membrane are involved in the pathogenesis of interferon nephrotoxicity, and that its mechanism of nephrotoxicity is similar to that of other low molecular weight proteins

  11. 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.

  12. Weight loss and elevated gluconeogenesis from alanine in lung cancer patients

    NARCIS (Netherlands)

    S. Leij-Halfwerk (Susanne); P.C. Dagnelie (Pieter); J.W.O. van den Berg (Willem); J.L.D. Wattimena (Josias); C.H. Hordijk-Luijk; J.H.P. Wilson (Paul)

    2000-01-01

    textabstractBACKGROUND: The role of gluconeogenesis from protein in the pathogenesis of weight loss in lung cancer is unclear. OBJECTIVE: Our aim was to study gluconeogenesis from alanine in lung cancer patients and to analyze its relation to the degree of weight loss.

  13. Selfcalibrated alanine/EPR dosimeters. A new generation of solid state/EPR dosimeters

    International Nuclear Information System (INIS)

    Yordanov, N.D.; Gancheva, V.

    1999-01-01

    Alanine/EPR dosimeters are well established as secondary, reference dosimeters for high-energy radiation. However, there are various sources of uncertainty in the evaluation of absorbed dose. This arises primarily from the necessity to calibrate each EPR spectrometer and each batch of dosimeters before their use. In order to overcome this disadvantage, a new generation alanine/EPR dosimeter has been developed, and its possibilities as a radiation detector are reported. Principally, it is a mixture of alanine, some quantity of EPR active substance, and a binding material. The EPR active substance, acting as an internal EPR standard, is chosen to have EPR parameters which are independent of the irradiation dose. The simultaneous recording of the spectra of both the sample and the standard under the same experimental conditions and the estimation of the ratio I alanine /I Mn as a function of the absorbed dose strongly reduces the uncertainties. The response of these dosimeters for 60 Co γ-radiation exhibits excellent linearity and reproducibility in the range of absorbed dose, 10 2 - 5 x 10 4 Gy. (author)

  14. Electron paramagnetic resonance radiation dosimetry: possible inorganic alternatives to the EPR/alanine dosimeter

    International Nuclear Information System (INIS)

    Keizer, P.N.; Morton, J.R.; Preston, K.F.

    1991-01-01

    The intensity of the EPR spectrum of γ-irradiated L-α-alanine has been accepted by the International Atomic Energy Agency as a secondary standard for high-dose (10-100 000 Gy) dosimetry. The alanine dosimeter is not without its disadvantages, however, and in this article alternative EPR dosimeters are explored. These include SO 3 - in irradiated K 2 CH 2 (SO 3 ) 2 and CO 2 - in irradiated sodium formate (NaHCO 2 ), both of which have some advantages over CH 3 CHCO 2 - in L-α-alanine. Using as a readout parameter the peak-to-peak excursion of the strongest line, these systems have a four-fold sensitivity advantage over alanine. The radicals SO 3 - and CO 2 - are, moreover, found in a wide variety of matrices, and it may be possible to find one in which they are even stronger. The need to discover a dosimeter material sensitive enough to function in the 'clinical' dose range (below 10 Gy) is emphasized. (author)

  15. New Generation of self-calibrated SS/EPR dosimeters: Alanine/EPR dosimeters

    International Nuclear Information System (INIS)

    Yordanov, N.D.; Gancheva, V.

    1999-01-01

    A new type of solid state/EPR dosimeters is described. Principally, it contains radiation sensitive diamagnetic material, some quantity of EPR active, but radiation insensitive, substance (for example Mn 2+ /MgO) and a binding material. In the present case alanine is used as a radiation sensitive substance. With this dosimeter, the EPR spectra of alanine and Mn 2+ are simultaneously recorded and the calibration graph represents the ratio of alanine versus Mn 2+ EPR signal intensity as a function of absorbed dose. In this way the reproducibility of the results is expected to be improved significantly including their intercomparison among different laboratories. Homogeneity of the prepared dosimeters and their behaviour (fading of EPR signals with time, influence of different meteorological conditions) show satisfactory reproducibility and stability with time. Because two different EPR active samples are recorded simultaneously, the influence of some instrument setting parameters (microwave power, modulation amplitude and modulation frequency) on the ratio I alanine /I Mn is also investigated. (author)

  16. Beta-alanine/alpha-ketoglutarate aminotransferase for 3-hydroxypropionic acid production

    Science.gov (United States)

    Jessen, Holly Jean; Liao, Hans H; Gort, Steven John; Selifonova, Olga V

    2014-11-18

    The present disclosure provides novel beta-alanine/alpha ketoglutarate aminotransferase nucleic acid and protein sequences having increased biological activity. Also provided are cells containing such enzymes, as well as methods of their use, for example to produce malonyl semialdehyde and downstream products thereof, such as 3-hydroxypropionic acid and derivatives thereof.

  17. Electronic structure and first hyperpolarizability of poly (μ2-L-alanine ...

    Indian Academy of Sciences (India)

    Poly(2-L-alanine-3-sodium nitrate (I)), -LASN, crystals have been grown by slow evaporation at room temperature. The nominal size of the crystals obtained by the method was of 500 nm. The UV–Vis spectrum shows a wide range, where absorption is lacking around 532 nm, which is required in order to have the ...

  18. Electronic structure and first hyperpolarizability of poly(μ2-L-alanine ...

    Indian Academy of Sciences (India)

    Administrator

    Abstract. Poly(μ2-L-alanine-μ3-sodium nitrate (I)), p-LASN, crystals have been grown by slow evaporation at room temperature. The nominal size of the crystals obtained by the method was of 500 nm. The UV–Vis spectrum shows a wide range, where absorption is lacking around 532 nm, which is required in order to have.

  19. Autolysis of Lactococcus lactis is increased upon D-alanine depletion of peptidoglycan and lipoteichoic acids

    NARCIS (Netherlands)

    Steen, Anton; Palumbo, Emmanuelle; Deghorain, Marie; Cocconcelli, Pier Sandro; Delcour, Jean; Kuipers, Oscar P.; Kok, Jan; Buist, Girbe; Hols, Pascal

    Mutations in the genes encoding enzymes responsible for the incorporation of D-Ala into the cell wall of Lactococcus lactis affect autolysis. An L. lactis alanine racemase (alr) mutant is strictly dependent on an external Supply Of D-Ala to be able to synthesize peptidoglycan and to incorporate

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

    Directory of Open Access Journals (Sweden)

    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.

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

    Science.gov (United States)

    Pey, Angel L.; Albert, Armando; Salido, Eduardo

    2013-01-01

    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. PMID:23956997

  2. 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.

  3. Follow-up of mild alanine aminotransferase elevation identifies hidden hepatitis C in primary care

    NARCIS (Netherlands)

    Helsper, C.W.|info:eu-repo/dai/nl/338910816; van Essen, G.A.|info:eu-repo/dai/nl/141200065; Frijling, B.D.; de Wit, N.J.|info:eu-repo/dai/nl/100525393

    2012-01-01

    BACKGROUND: Hepatitis C (HCV) and hepatitis B (HBV) virus infection can lead to serious complications if left untreated, but often remain undetected in primary care. Mild alanine aminotransferase (ALT) elevations (30-100 IU/l) are commonly found and could be associated with viral hepatitis;

  4. Electronic structure and first hyperpolarizability of poly(μ2-L-alanine ...

    Indian Academy of Sciences (India)

    Administrator

    poly(μ2-L-alanine-μ3-sodium nitrate (I)) crystals. A DUARTE MOLLER. Centro de Investigación en Materiales Avanzados, SC, Miguel de Cervantes 120, Complejo Industrial Chihuahua,. Chihuahua 31109, México. Present address: Universidad Tecnológica de Querétaro, Avenida Pie de la Cuesta 2501, Unidad Nacional,.

  5. Recent progress in application of JAERI alanine/ESR dosimetry system

    International Nuclear Information System (INIS)

    Kojima, T.

    1995-01-01

    Feasibility studies of application of JAERI alanine/ESR dosimetry system were performed on radiotherapy level dosimetry, low dose-rate dosimetry for residual life estimation of cable insulators used in nuclear power facilities, and dose monitoring for electron processing. (author)

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

    Science.gov (United States)

    Coqueiro, Audrey Yule; Raizel, Raquel; Bonvini, Andrea; Hypólito, Thaís; Godois, Allan da Mata; Pereira, Jéssica Ramos Rocha; Garcia, Amanda Beatriz de Oliveira; Lara, Rafael de Souza Bittencourt; Rogero, Marcelo Macedo; Tirapegui, Julio

    2018-01-25

    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.

  7. Efeito do Exercício com 1 e 3 Minutos de Intervalo de Descanso Entre as Séries na Atividade Sérica das Transaminases

    Directory of Open Access Journals (Sweden)

    Ramon Martins Alves

    2011-12-01

    Full Text Available O treinamento de força (TF intenso ocasiona danos na estrutura muscular, que podem ser detectados por meio de marcadores indiretos. OBJETIVO. Verificar se há alterações na atividade da creatina quinase (CK, da aspartato transaminase (AST e alanina transaminase (ALT com o TF em diferentes intervalos entre séries e exercícios. MÉTODOS. Treze voluntários (± 21,8 anos; ±173,6 cm; ±68,3 kg realizaram o teste de uma repetição máxima (1RM nos exercícios: supino reto, puxada (pulley, desenvolvimento sentado, tríceps (pulley e rosca bíceps. Após sete dias, todos foram submetidos a uma coleta de sangue (PRE e realizaram 4 séries máximas até a falha concêntrica com intensidade de 85% de 1RM e intervalos de 1 ou 3 minutos entre as séries e exercícios. Todos retornaram ao mesmo local 24, 48 e 72 horas após a sessão para mais uma coleta de sangue. Repetindo os procedimentos após uma semana, porém os voluntários que realizaram o teste com intervalo de 1 minuto na semana seguinte manteriam intervalos de 3 minutos e vice-versa. Realizou-se ANOVA mantendo como significância p<0,05. RESULTADOS. Houve redução significativa do número de repetições a cada série de todos os exercícios com 1 e 3 minutos de intervalo. Observou-se um aumento significativo da atividade sérica de CK e de AST 24, 48 e 72 horas utilizando ambos os intervalos. ALT não se alterou. CONCLUSÃO. O TF causou aumento na atividade de CK e AST, mas sem apresentar diferenças com os intervalos utilizados. Porém, pequenos intervalos reduzem o volume total de repetições.

  8. Knockout of the alanine racemase gene in Aeromonas hydrophila HBNUAh01 results in cell wall damage and enhanced membrane permeability.

    Science.gov (United States)

    Liu, Dong; Zhang, Lu; Xue, Wen; Wang, Yaping; Ju, Jiansong; Zhao, Baohua

    2015-07-01

    This study focused on the alanine racemase gene (alr-2), which is involved in the synthesis of d-alanine that forms the backbone of the cell wall. A stable alr-2 knockout mutant of Aeromonas hydrophila HBNUAh01 was constructed. When the mutant was supplemented with d-alanine, growth was unaffected; deprivation of d-alanine caused the growth arrest of the starved mutant cells, but not cell lysis. No alanine racemase activity was detected in the culture of the mutant. Additionally, a membrane permeability assay showed increasing damage to the cell wall during d-alanine starvation. No such damage was observed in the wild type during culture. Scanning and transmission electron microscopy analyses revealed deficiencies of the cell envelope and perforation of the cell wall. Leakage of UV-absorbing substances from the mutants was also observed. Thus, the partial viability of the mutants and their independence of d-alanine for growth indicated that inactivation of alr-2 does not impose an auxotrophic requirement for d-alanine. © FEMS 2015. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  9. 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

  10. Effect of beta-alanine and carnosine supplementation on muscle contractility in mice.

    Science.gov (United States)

    Everaert, Inge; Stegen, Sanne; Vanheel, Bert; Taes, Youri; Derave, Wim

    2013-01-01

    Enhanced carnosine levels have been shown to be ergogenic for high-intensity exercise performances, although the role of carnosine in the control of muscle function is poorly understood. Therefore, the aim of this study was to investigate the effect of long-term supplementation with increasing doses of carnosine and beta-alanine on muscle carnosine, anserine, and taurine levels and on in vitro contractility and fatigue in mice. Male Naval Medical Research Institute mice (n = 66) were control fed or supplemented with either carnosine (0.1%, 0.5%, or 1.8%) or beta-alanine (0.6 or 1.2%) in their drinking water for 8-12 wk. Soleus and extensor digitorum longus (EDL) were tested for in vitro contractile properties, and carnosine, anserine, and taurine content were measured in EDL and tibialis anterior by high-performance liquid chromatography. Only supplementation with 1.8% carnosine and 1.2% beta-alanine resulted in markedly higher carnosine (up to +160%) and anserine levels (up to +46%) compared with control mice. Beta-alanine supplementation (1.2%) resulted in increased fatigue resistance in the beginning of the fatigue protocol in soleus (+2%-4%) and a marked leftward shift of the force-frequency relation in EDL (10%-31% higher relative forces). Comparable with humans, beta-alanine availability seems to be the rate-limiting step for synthesis of muscle histidine-containing dipeptides in mice. Moreover, muscle histidine-containing dipeptides loading in mice moderately and muscle dependently affects excitation-contraction coupling and fatigue.

  11. K-band EPR dosimetry: small-field beam profile determination with miniature alanine dosimeter

    International Nuclear Information System (INIS)

    Chen, Felipe; Graeff, Carlos F.O.; Baffa, Oswaldo

    2005-01-01

    The use of small-size alanine dosimeters presents a challenge because the signal intensity is less than the spectrometer sensitivity. K-band (24 GHz) EPR spectrometer seems to be a good compromise between size and sensitivity of the sample. Miniature alanine pellets were evaluated for small-field radiation dosimetry. Dosimeters of DL-alanine/PVC with dimensions of 1.5 mm diameter and 2.5 mm length with 5 mg mass were developed. These dosimeters were irradiated with 10 MV X-rays in the dose range 0.05-60 Gy and the first harmonic (1 h) spectra were recorded. Microwave power, frequency and amplitude of modulation were optimized to obtain the best signal-to-noise ratio (S/N). For beam profile determination, a group of 25 dosimeters were placed in an acrylic device with dimensions of (7.5x2.5x1) cm 3 and irradiated with a (3x3) cm 2 10 MV X-rays beam field size. The dose at the central region of the beam was 20 Gy at a depth of 2.2 cm (build up for acrylic). The acrylic device was oriented perpendicular to the beam axis and to the gantry rotation axis. For the purposes of comparison of the spatial resolution, the beam profile was also determined with a radiographic film and 2 mm aperture optical densitometer; in this case the dose was 1 cGy. The results showed a similar spatial resolution for both types of dosimeters. The dispersion in dose reading was larger for alanine in comparison with the film, but alanine dosimeters can be read faster and more directly than film over a wide dose range

  12. 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.

  13. [Evaluation of hepatic gluconeogenesis by intravenous L-alanine tolerance test after major hepatectomy].

    Science.gov (United States)

    Miyagi, N

    1989-11-01

    The response of glucose, insulin and glucagon to L-alanine tolerance test (0.5 g/kg, intravenously, A.T.T.) was examined in dogs following hepatectomy. Before A.T.T., dogs were starved for 24 hours after antecedent carbohydrate-free diets. Under this condition, hepatic glycogen and blood glucose were 0.5 +/- 0.2 mg/g liver and 85 +/- 11.9 mg/dl, respectively. The marked response of blood glucose to alanine tolerance test was observed and the maximal increment above the basal level was 47 +/- 5.2 mg/dl at 30 minutes after A.T.T. One week after 44% and 70% hepatectomy, the increments of glucose at 30 minutes after A.T.T. were 58% and 24% respectively, as compared with those before hepatectomy. These results suggest that the glucose response to alanine tolerance is useful for an index of functional reserve of glucose production in the remnant liver after hepatectomy. The increments of blood glucose at 30 minutes after A.T.T. 1, 2, 3 and 4 weeks after 70% hepatectomy were 24.0, 21.8, 52.7 and 80% of that before hepatectomy respectively. The insulin response to alanine tolerance gave two peaks at 2.5 min and 30 min and the latter peak was dependent on the blood glucose response before and after hepatectomy. One or two weeks after 70% hepatectomy, the exaggerated glucagon response to alanine tolerance was observed. These results suggest that A.T.T. is not only an excellent index of functional reserve of hepatic gluconeogenesis, but also is an appropriate method for investigation of the relation between endogenous glucose production and endocrine pancreas.

  14. 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.

  15. The metabolism of histamine in the Drosophila optic lobe involves an ommatidial pathway: β-alanine recycles through the retina

    Science.gov (United States)

    Borycz, Janusz; Borycz, Jolanta A.; Edwards, Tara N.; Boulianne, Gabrielle L.; Meinertzhagen, Ian A.

    2012-01-01

    SUMMARY Flies recycle the photoreceptor neurotransmitter histamine by conjugating it to β-alanine to form β-alanyl-histamine (carcinine). The conjugation is regulated by Ebony, while Tan hydrolyses carcinine, releasing histamine and β-alanine. In Drosophila, β-alanine synthesis occurs either from uracil or from the decarboxylation of aspartate but detailed roles for the enzymes responsible remain unclear. Immunohistochemically detected β-alanine is present throughout the fly’s entire brain, and is enhanced in the retina especially in the pseudocone, pigment and photoreceptor cells of the ommatidia. HPLC determinations reveal 10.7 ng of β-alanine in the wild-type head, roughly five times more than histamine. When wild-type flies drink uracil their head β-alanine increases more than after drinking l-aspartic acid, indicating the effectiveness of the uracil pathway. Mutants of black, which lack aspartate decarboxylase, cannot synthesize β-alanine from l-aspartate but can still synthesize it efficiently from uracil. Our findings demonstrate a novel function for pigment cells, which not only screen ommatidia from stray light but also store and transport β-alanine and carcinine. This role is consistent with a β-alanine-dependent histamine recycling pathway occurring not only in the photoreceptor terminals in the lamina neuropile, where carcinine occurs in marginal glia, but vertically via a long pathway that involves the retina. The lamina’s marginal glia are also a hub involved in the storage and/or disposal of carcinine and β-alanine. PMID:22442379

  16. NÍVEIS SÉRICOS DE TRANSAMINASE GLUTÂMICA-OXALACÉTICA (GOT E TRANSAMINASE GLUTÂMICA-PIRÚVICA (GPT EM BOVINOS DO ESTADO DE GOIÁS SERUM LEVELS OF GOT AND GPT IN BOVINES IN THE GOIÁS STATE

    Directory of Open Access Journals (Sweden)

    Eduardo Cavalheiro Jardim

    2007-09-01

    Full Text Available

    Foram analisados 162 hemossoros de bovinos mestiços zebu com a finalidade de obtenção dos níveis séricos da transaminase glutâmica-oxalacética e da transaminase glutâmica-pirúvica, de 18 bezerros(as, 31 novilhos(as e 83 vacas, procedentes de municípios do Estado de Goiás. Os valores encontrados foram os seguintes (URF: TGO, 77,5 +/- 27,3 (bezerros(as, 72,9 +/- 19,6 (novilhos(as e 81,8 +/- 28,4 (vacas; TGP, 29,5 +/- 8,1 (novilhos(as, 31,1 +/- 8.3 (novilhos(as e 34,1 +/- 10,6 (vacas. É muito difícil estandadizar condições de manejo, e os dados encontrados no presente trabalho confirmam que as transaminases são enzimas, dinâmicas, capazes de responderem rapidamente a um grande número de trocas fisiológicas. Posteriores trabalhos devem ser desenvolvidos buscando uniformizar os grupos de trabalho sob os diferentes pontos considerados no presente estudo.

    With the objective of studying the serum levels of GPT and GOT, 162 blood serum from crossbred zebu were analyzed. The blood was taken from 48 calves, 31 steers and 83 cows from farms of counties in the State of Goiás. GOT serum levels (URF were as fo1lows: calves, 72.5 +/- 27.3, steers, 72.9 +/- 19.6 and cows 81.8 +/-28.4. For GPT serum levels (URF was found: calves, 29.5 +/- 8.1, steers, 31.1 +/- 8.3 and cows 34.1 +/- 10.6. Some differences were found in GOT and GPT, in comparison with other foreign reports in mature cows.

  17. The alanine detector in BNCT dosimetry: dose response in thermal and epithermal neutron fields.

    Science.gov (United States)

    Schmitz, T; Bassler, N; Blaickner, M; Ziegner, M; Hsiao, M C; Liu, Y H; Koivunoro, H; Auterinen, I; Serén, T; Kotiluoto, P; Palmans, H; Sharpe, P; Langguth, P; Hampel, G

    2015-01-01

    The response of alanine solid state dosimeters to ionizing radiation strongly depends on particle type and energy. Due to nuclear interactions, neutron fields usually also consist of secondary particles such as photons and protons of diverse energies. Various experiments have been carried out in three different neutron beams to explore the alanine dose response behavior and to validate model predictions. Additionally, application in medical neutron fields for boron neutron capture therapy is discussed. Alanine detectors have been irradiated in the thermal neutron field of the research reactor TRIGA Mainz, Germany, in five experimental conditions, generating different secondary particle spectra. Further irradiations have been made in the epithermal neutron beams at the research reactors FiR 1 in Helsinki, Finland, and Tsing Hua open pool reactor in HsinChu, Taiwan ROC. Readout has been performed with electron spin resonance spectrometry with reference to an absorbed dose standard in a (60)Co gamma ray beam. Absorbed doses and dose components have been calculated using the Monte Carlo codes fluka and mcnp. The relative effectiveness (RE), linking absorbed dose and detector response, has been calculated using the Hansen & Olsen alanine response model. The measured dose response of the alanine detector in the different experiments has been evaluated and compared to model predictions. Therefore, a relative effectiveness has been calculated for each dose component, accounting for its dependence on particle type and energy. Agreement within 5% between model and measurement has been achieved for most irradiated detectors. Significant differences have been observed in response behavior between thermal and epithermal neutron fields, especially regarding dose composition and depth dose curves. The calculated dose components could be verified with the experimental results in the different primary and secondary particle fields. The alanine detector can be used without

  18. 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.

  19. New classes of alanine racemase inhibitors identified by high-throughput screening show antimicrobial activity against Mycobacterium tuberculosis.

    Directory of Open Access Journals (Sweden)

    Karen G Anthony

    Full Text Available In an effort to discover new drugs to treat tuberculosis (TB we chose alanine racemase as the target of our drug discovery efforts. In Mycobacterium tuberculosis, the causative agent of TB, alanine racemase plays an essential role in cell wall synthesis as it racemizes L-alanine into D-alanine, a key building block in the biosynthesis of peptidoglycan. Good antimicrobial effects have been achieved by inhibition of this enzyme with suicide substrates, but the clinical utility of this class of inhibitors is limited due to their lack of target specificity and toxicity. Therefore, inhibitors that are not substrate analogs and that act through different mechanisms of enzyme inhibition are necessary for therapeutic development for this drug target.To obtain non-substrate alanine racemase inhibitors, we developed a high-throughput screening platform and screened 53,000 small molecule compounds for enzyme-specific inhibitors. We examined the 'hits' for structural novelty, antimicrobial activity against M. tuberculosis, general cellular cytotoxicity, and mechanism of enzyme inhibition. We identified seventeen novel non-substrate alanine racemase inhibitors that are structurally different than any currently known enzyme inhibitors. Seven of these are active against M. tuberculosis and minimally cytotoxic against mammalian cells.This study highlights the feasibility of obtaining novel alanine racemase inhibitor lead compounds by high-throughput screening for development of new anti-TB agents.

  20. Thermochemistry of the solution of β-alanine in (H2O + alcohol) mixtures at 298.15 K

    International Nuclear Information System (INIS)

    Smirnov, Valeriy I.; Badelin, Valentin G.

    2013-01-01

    Highlights: • Enthalpies of β-alanine dissolution have been measured in aqueous solution of MeOH, EtOH, 1-PrOH and 2-PrOH. • Measured data were reported as functions of composition of water + alcohol mixtures. • Enthalpy coefficients of pairwise interactions have been analyzed in terms of McMillan–Mayer theory. - Abstract: The enthalpies of the solution of β-alanine in H 2 O + (methanol, ethanol, 1-propanol and 2-propanol) mixtures with alcohol content up to 0.4 mol fractions, have been determined calorimetrically at T = 298.15 K. The standard enthalpies of the solution and transfer of β-alanine from water to aqueous alcohol have been calculated. The effect of structure properties of a mixed solvent on specified enthalpy characteristics of β-alanine is discussed. The enthalpy coefficients of pairwise interactions between β-alanine and alcohol molecules have been computed. It has been found that these coefficients become increasingly positive in methanol, ethanol, 1-propanol, and 2-propanol sequence. A comparative analysis of thermodynamic characteristics of dissolution of β-alanine and D,L-α-alanine in the mixtures studied has been made

  1. Calculations of the relative effectiveness of alanine for neutrons with energies up to 17.1 MeV

    International Nuclear Information System (INIS)

    Gerstenberg, H.M.; Coyne, J.J.

    1990-01-01

    The relative effectiveness (RE) of alanine has been calculated for neutrons using the RE of alanine for charged particles. The neutrons interact with one or more of the elements (hydrogen, carbon, nitrogen and oxygen) that compose the alanine. These interactions produce spectra of secondary charged particles consisting of ions of H, D, He, Be, B, C, N and O. From a combination of the calculated secondary charged particle spectra generated by the slowing down neutrons, and the calculated RE of the ions produced, a RE for the neutrons can be obtained. In addition, lineal energy spectra were determined for neutrons with energies up to 17.1 MeV interacting with alanine. An analytical code was used to calculate these spectra for a 1 μm diameter alanine cell surrounded by an alanine medium. For comparison, similar calculations were made for muscle tissue. Finally, the calculated differential RE was folded with dose distributions to obtain RE-weighted distributions for alanine. (author)

  2. Dosimetry auditing procedure with alanine dosimeters for light ion beam therapy

    DEFF Research Database (Denmark)

    Ableitinger, Alexander; Vatnitsky, Stanislav; Herrmann, Rochus

    2013-01-01

    Background and purpose In the next few years the number of facilities providing ion beam therapy with scanning beams will increase. An auditing process based on an end-to-end test (including CT imaging, planning and dose delivery) could help new ion therapy centres to validate their entire logistic...... of the biological dose is out of scope of the current work. Materials and methods The audit procedure was based on a homogeneous phantom that mimics the dimension of a head (20 × 20 × 21 cm3). The phantom can be loaded either with an ionisation chamber or 20 alanine dosimeters plus 2 radiochromic EBT films. Dose...... of about 3%. Conclusions Alanine dosimeters are suitable detectors for dosimetry audits in ion beam therapy and the presented end-to-end test is feasible. If further studies show similar results, this dosimetric audit could be implemented as a credentialing procedure for clinical proton and carbon beam...

  3. Development of DL-alanine systems for gamma radiation and electron dosimetry

    International Nuclear Information System (INIS)

    Costa, Zelia Maria da

    1994-01-01

    Two different dosimetric systems using DL-Alanine samples were employed to determine the absorbed dose from 60 Co gamma-rays source and electrons emitted from an accelerator. The first dosimetric system is based on the relationship between free radicals produced and the absorbed dose using the electron spin resonance (ESR) technique. Details on the sample preparation, the spectrometer parameter setting, the analysis of the ESR signal to dose, the influence of dose rate and the radiation type dependence are described. The second dosimetric system is based on the determination by absorbance spectrophotometry of the complex produced, which are formed when the irradiated alanine is dissolved in a solution containing ferrous ammonium sulphate xylenol in 0,05 N H 2 SO 4 . Different concentrations for each reagents has been analyzed in the preparation of this solution as well as the influence caused by radiation type and dose rate in the absorbance. (author)

  4. Utilization of ALANPOL dosemeter composed by alanine and polyethylene for determination of doses in industrial effluents

    International Nuclear Information System (INIS)

    Rela, Carolina S.; Sampa, Maria Helena de O.; Napolitano, Celia M.; Pontuschka, Walter M.

    2005-01-01

    A study of traceability and accuracy was performed with the dosimetry system ALANPOL developed by IVHTJ/ Poland that uses alanine as a dosimeter and EPR as the read out system. The dosimeter ALANPOL is a polymer compound that aggregates alanine. The dosimeter has the outside diameter 3 mm, and the length of 25,4 mm (1 inch). In order to use this dosimeter for monitoring the existing real time calorimetry dosimeter system set up in a electron beam waste treatment plant. Electron beam and gamma irradiation experiments with ALANPOL was carried out under demineralized water and industrial effluent. The result show that in the dose range of 1-40 kGy the ALANPOL dosimeter system is suitable underwater monitoring for absorbed dose and can be applied for the evaluation of the on line calorimetry dosimeter system. (author)

  5. Braked rotation of CH3 group in L-alanine monocrystals: temperature transformation of EPR spectrum

    International Nuclear Information System (INIS)

    Lemanov, V.V.; Sochava, L.S.

    2003-01-01

    EPR spectra temperature transformation of the irradiated alanine crystals is used for studying rotation of CH 3 methyl group in L-alamine monocrystals. 60 Co (2 x 10 4 Gy dose) was applied as a γ-radiation source. The simple method of experimental data processing which is reduced to obtaining the resonance lines width dependence on the temperature is used for the quantitative analysis of the spectrum temperature transformation. Temperature dependence of the CH 3 group rotation frequency is identified on the basis of these data. Activation energy U = 0.18 eV and pre-exponential multiplier ω 0 = 10 13 s -1 are determined from the EPR spectra temperature transformation which are in good agreement with values obtained earlier from the measurements of the proton spin-lattice relaxation in alanine polycrystal samples [ru

  6. A reference dosimetric system for dose interval of radiotherapy based on alanine/RPE

    International Nuclear Information System (INIS)

    Rodrigues Junior, Orlando; Galante, Ocimar L.; Campos, Leticia L.

    2001-01-01

    This work describes the development of a reference dosimetric system based on alanine/EPR for radiotherapy dose levels. Currently the IPEN is concluding a similar system for the dose range used for irradiation of products, 10-10 5 Gy. The objective of this work is to present the efforts towards to improve the measure accuracy for doses in the range between 1-10 Gy. This system could be used as reference by radiotherapy services, as much in the quality control of the equipment, as for routine accompaniment of more complex handling where the total doses can reach some grays. The system uses alanine as detector and electronic paramagnetic resonance - EPR as measure technique. To reach accuracy better than 5% mathematical studies on the best optimization of the EPR spectrometer parameters and methods for the handling of the EPR sign are discussed. (author)

  7. Naturally Inspired Peptide Leads: Alanine Scanning Reveals an Actin-Targeting Thiazole Analogue of Bisebromoamide.

    Science.gov (United States)

    Johnston, Heather J; Boys, Sarah K; Makda, Ashraff; Carragher, Neil O; Hulme, Alison N

    2016-09-02

    Systematic alanine scanning of the linear peptide bisebromoamide (BBA), isolated from a marine cyanobacterium, was enabled by solid-phase peptide synthesis of thiazole analogues. The analogues have comparable cytotoxicity (nanomolar) to that of BBA, and cellular morphology assays indicated that they target the actin cytoskeleton. Pathway inhibition in human colon tumour (HCT116) cells was explored by reverse phase protein array (RPPA) analysis, which showed a dose-dependent response in IRS-1 expression. Alanine scanning reveals a structural dependence to the cytotoxicity, actin targeting and pathway inhibition, and allows a new readily synthesised lead to be proposed. © 2016 The Authors. Published by Wiley-VCH Verlag GmbH & Co. KGaA.

  8. Orientation of crystals in alanine dosimeter assessed by DRS, as seen in EPR spectra evaluation

    International Nuclear Information System (INIS)

    Grazyna Przybytniak; Zagorski, Z.P.

    1996-01-01

    The alanine dosimeter made for evaluation by diffuse light reflection spectrophotometry (ALA/DRS) does not show the effect of orientation of crystals. Supposed deviation from random orientation has been investigated by EPR spectroscopy. EPR investigation shows that in spite of the very fine size of L-alanine crystals, they are oriented in thin layers of the polyethylene matrix. Specially prepared films with deliberately well oriented crystals have confirmed this observation. Our ALA/DRS dosimeter can be evaluated by the EPR method for the concentration of free radicals, providing that the dominating crystal orientation in the dosimetric film is indicated on it as an arrow, and the sample is inserted into the magnetic cavity always in the same orientation as has been done during the calibration operation. (author). 6 refs., 2 figs

  9. 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)

  10. Synthesis, Characterization and Metal Ion Detection of Novel Fluoroionophores Based on Heterocyclic Substituted Alanines

    Science.gov (United States)

    Costa, Susana P. G.; Oliveira, Elisabete; Lodeiro, Carlos; Raposo, M. Manuela M.

    2007-01-01

    The synthesis of new fluorescent probes containing the thiophene and benzoxazole moieties combined with an alanine residue is described. The resulting highly fluorescent heterocyclic alanine derivatives respond via a quenching effect, with paramagnetic Cu(II) and Ni(II) metal ions and with diamagnetic Hg(II), as shown by the absorption and steady-state fluorescence spectroscopy studies. The formation of mononuclear or dinuclear metal complexes was postulated based on the presence of the free carboxylic acid as binding site and also with the interaction with the donor atoms in the chromophore. Interaction with other important biological metal ions such as Zn(II), Ca(II) and Na(I) was also explored. PMID:28903216

  11. Volume dose ratios relevant for alanine dosimetry in small, 6 MV photon beams

    DEFF Research Database (Denmark)

    Cronholm, Rickard O.; Andersen, Claus Erik; Behrens, Claus F.

    2012-01-01

    therapy). To this end, we here present the results of a Monte Carlo simulation study with DOSRZnrc that investigated the influence of field and detector size for small 6 MV photon beams. The study focusses on doses averaged over the volume of the detector rather than point doses.The ratio of volume...... averaged doses to water (D¯W) and alanine (D¯det) was found to be approximately 1.025 for most situations studied, and a constant ratio is likely to be representative for many applications in radiation therapy. However, D¯W/D¯det was found to be as low as 0.9908 ± 0.0037 in situations where one might...... expect significant deviations from charged particle equilibrium (i.e. at shallow depths and when the field size was smaller than the range of the secondary electrons). These effects therefore need consideration when finite-size alanine dosimeters are used under such conditions....

  12. Unusual hydroxyl migration in the fragmentation of β-alanine dication in the gas phase.

    Science.gov (United States)

    Piekarski, Dariusz Grzegorz; Delaunay, Rudy; Maclot, Sylvain; Adoui, Lamri; Martín, Fernando; Alcamí, Manuel; Huber, Bernd A; Rousseau, Patrick; Domaracka, Alicja; Díaz-Tendero, Sergio

    2015-07-14

    We present a combined experimental and theoretical study of the fragmentation of doubly positively charged β-alanine molecules in the gas phase. The dissociation of the produced dicationic molecules, induced by low-energy ion collisions, is analysed by coincidence mass spectrometric techniques; the coupling with ab initio molecular dynamics simulations allows rationalisation of the experimental observations. The present strategy gives deeper insights into the chemical mechanisms of multiply charged amino acids in the gas phase. In the case of the β-alanine dication, in addition to the expected Coulomb explosion and hydrogen migration processes, we have found evidence of hydroxyl-group migration, which leads to unusual fragmentation products, such as hydroxymethyl cation, and is necessary to explain some of the observed dominant channels.

  13. 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

  14. A comparative study of the metal binding behavior of alanine based bis-thiourea isomers

    OpenAIRE

    Fakhar, Imran; Yamin, Bohari M.; Hasbullah, Siti Aishah

    2017-01-01

    Two new symmetrical bis-thiourea, 2,2?-[{(terephthaloylbis(azanediyl)bis(carbonothioyl) bis(azanediyl)}dipropanoic acid] (1A) and 3,3?-[{(terephthaloylbis(azanediyl)bis (carbonothioyl)bis(azanediyl)} dipropanoic acid] (1B) were synthesized by the reaction of terephthaloyl chloride with ?- and ?-alanine in good yields. Their binding properties were investigated with various metal cations using UV?Vis titration experiments. Both isomers exhibited effective binding with Ag+, Cu2+, Hg2+, Pb2+, Fe...

  15. Peptide conformational preferences in osmolyte solutions: Transfer free energies of deca-alanine

    OpenAIRE

    Kokubo, Hironori; Hu, Char Y.; Pettitt, B. Montgomery

    2011-01-01

    The nature in which the protecting osmolyte trimethylamine-N-oxide (TMAO) and the denaturing osmolyte urea affect protein stability is investigated simulating a deca-alanine peptide model in multiple conformations of the denatured ensemble. Binary solutions of both osmolytes and mixed osmolyte solutions at physiologically-relevant concentrations of 2:1 (urea:TMAO) are studied using standard molecular dynamics simulations and solvation free energy calculations. Component analysis reveals the d...

  16. An Exonic Insertion Encodes an Alanine Stretch in Porcine Synapsin I

    DEFF Research Database (Denmark)

    Hedegaard, Claus; Bendixen, Emøke; Jensen, Poul Henning

    2009-01-01

    The synapsin (syn) protein family comprises the neuron-specific phosphoproteins syn I, II, and III, all found in at least two isoforms (Südhof et al. 1989 ; Porton et al. 1999 ). Mature syns localize to the membrane of neurotransmitter-releasing synaptic vesicles; they are expected to have regula......, including identification and verification at the protein level of an alanine-encoding insert...

  17. Uracil and beta-alanine degradation in Saccharomyces Kluyveri - discovery of a novel catabolic pathway

    DEFF Research Database (Denmark)

    Andersen, Gorm

    2006-01-01

    ’en i gær og de genetiske forudsætninger for uracil og beta-alanine (BAL) katabolisme i S. kluyveri undersøgt. Evnen til at bruge uracil, dihydrouracil (DHU), beta-ureidopropionate (BUP) og BAL som nitrogenkilde blev studeret i 38 gær arter. Disse var udvalgt, så de dækkede “Saccharomyces komplekset...

  18. A COMPARATIVE STUDY ON THE ACTIVITY OF ALANIN-AMINOTRANSFERASE IN HYPOPHTHALMICHTHYS MOLITRIX AND ARISTICHTHYS NOBILIS

    Directory of Open Access Journals (Sweden)

    Gabriela Vasile

    2006-08-01

    Full Text Available The present paper represents a comparative study on the activity of one aminotransferase - alaninaminotransferase, in the digestive tube of Hypophthalmichthys molitrix (silver carp and Aristichthys nobilis (bighead carp. The enzymatic activity has been determined colorimetrically, with 2, 4 - dinitrophenyl hydrazine, the results obtained being expressed as UE / g / min. It was observed that, comparatively with the alanin-aminotransferase activity recorded in silver carp, in the case of bighead carp, the values recorded are much lower.

  19. Mechanism of leukotriene D4 inhibition of Na-alanine cotransport in intestinal epithelial cells

    OpenAIRE

    Talukder, Jamilur R.; Kekuda, Ramesh; Saha, Prosenjit; Sundaram, Uma

    2008-01-01

    In a rabbit model of chronic intestinal inflammation, we previously demonstrated inhibition of neutral Na-amino acid cotransport. The mechanism of the inhibition was secondary to a decrease in the affinity for amino acid rather than the number of cotransporters. Since leukotriene (LT)D4 is known to be elevated in enterocytes during chronic intestinal inflammation, we used rat intestinal epithelial cell (IEC-18) monolayers to determine the mechanism of regulation of Na-alanine cotransport (ala...

  20. β-alanine supplementation improves isometric endurance of the knee extensor muscles

    Directory of Open Access Journals (Sweden)

    Sale Craig

    2012-06-01

    Full Text Available Abstract Background We examined the effect of four weeks of β-alanine supplementation on isometric endurance of the knee extensors at 45% maximal voluntary isometric contraction (MVIC. Methods Thirteen males (age 23 ± 6 y; height 1.80 ± 0.05 m; body mass 81.0 ± 10.5 kg, matched for pre-supplementation isometric endurance, were allocated to either a placebo (n = 6 or β-alanine (n = 7; 6.4 g·d-1 over 4 weeks supplementation group. Participants completed an isometric knee extension test (IKET to fatigue, at an intensity of 45% MVIC, before and after supplementation. In addition, two habituation tests were completed in the week prior to the pre-supplementation test and a further practice test was completed in the week prior to the post-supplementation test. MVIC force, IKET hold-time, and impulse generated were recorded. Results IKET hold-time increased by 9.7 ± 9.4 s (13.2% and impulse by 3.7 ± 1.3 kN·s-1 (13.9% following β-alanine supplementation. These changes were significantly greater than those in the placebo group (IKET: t(11 = 2.9, p ≤0.05; impulse: t(11 = 3.1, p ≤ 0.05. There were no significant changes in MVIC force in either group. Conclusion Four weeks of β-alanine supplementation at 6.4 g·d-1 improved endurance capacity of the knee extensors at 45% MVIC, which most likely results from improved pH regulation within the muscle cell as a result of elevated muscle carnosine levels.

  1. Aspartate Aminotransferase (AST/GOT) and Alanine Aminotransferase (ALT/GPT) Detection Techniques

    OpenAIRE

    Huang, Xing-Jiu; Choi, Yang-Kyu; Im, Hyung-Soon; Yarimaga, Oktay; Yoon, Euisik; Kim, Hak-Sung

    2006-01-01

    The levels of aspartate aminotransferase (AST/GOT) and alanine aminotransferase (ALT/GPT) in serum can help people diagnose body tissues especially the heart and the liver are injured or not. This article provides a comprehensive review of research activities that concentrate on AST/GOT and ALT/GPT detection techniques due to their clinical importance. The detection techniques include colorimetric, spectrophotometric, chemiluminescence, chromatography, fluorescence and UV absorbance, radioche...

  2. Effects of inhibitors of GABA-transaminase on hole-board exploration and on temperature. Relation with effects on quasi-morphine abstinence behaviour induced by sodium dipropylacetate.

    Science.gov (United States)

    van der Laan, J W; Weick, G; van Bleek, G

    1985-11-01

    Four inhibitors of gamma-aminobutyric acid transaminase (GABA-T) were investigated together with respect to their effects on hole-board exploration and temperature and the relation with effects on quasi-morphine-abstinence behaviour induced by dipropylacetate (DPA) in rats. Amino-oxyacetic acid (AOAA), gamma-acetylenic-GABA (GAG), gamma-vinyl-GABA (GVC) and ethanolamine-O-sulfate (EOS) were found to reduce hole-board exploration especially in the higher doses used, although the time-course of the effect was different for the compounds. For EOS and GVG the decrease in hole-board exploration paralleled a strong hypothermic effect. The compounds AOAA and GAG exerted a less and more transient hypothermic effect. However, the decrease in hole-board exploration did not fall in with this decrease in temperature. AOAA and GAG were found to decrease DPA-induced body shakes and locomotor activity, while GVG and EOS had no effect on body shakes and transient effects but opposite to each other, on locomotor activity. The efficacy of the GABA-T-inhibitors was measured biochemically, and the influence on the activity of glutamate decarboxylase (GAD) was also determined. AOAA and GAG were found to be strong inhibitors of GABA-T whereas the other two compounds were less efficient in the used doses. In addition AOAA and GAG influenced the activity of GAD strongly, while using GVG only a small decrease was found. The results suggest that the anti-quasi-withdrawal, the sedative and the hypothermic effects are not related to each other nor related to an effect on GABA-T. The suppressive effects on quasi-withdrawal body shakes, however, could be related to the inhibition of GAD and a hypothesis involving a compartmentalized action of DPA on GABA-metabolism has been proposed.

  3. The periplasmic transaminase PtaA of Pseudomonas fluorescens converts the glutamic acid residue at the pyoverdine fluorophore to α-ketoglutaric acid.

    Science.gov (United States)

    Ringel, Michael T; Dräger, Gerald; Brüser, Thomas

    2017-11-10

    The periplasmic conversion of ferribactin to pyoverdine is essential for siderophore biogenesis in fluorescent pseudomonads, such as pathogenic Pseudomonas aeruginosa or plant growth-promoting Pseudomonas fluorescens The non-ribosomal peptide ferribactin undergoes cyclizations and oxidations that result in the fluorophore, and a strictly conserved fluorophore-bound glutamic acid residue is converted to a range of variants, including succinamide, succinic acid, and α-ketoglutaric acid residues. We recently discovered that the pyridoxal phosphate-containing enzyme PvdN is responsible for the generation of the succinamide, which can be hydrolyzed to succinic acid. Based on this, a distinct unknown enzyme was postulated to be responsible for the conversion of the glutamic acid to α-ketoglutaric acid. Here we report the identification and characterization of this enzyme in P. fluorescens strain A506. In silico analyses indicated a periplasmic transaminase in fluorescent pseudomonads and other proteobacteria that we termed PtaA for " p eriplasmic t ransaminase A " An in-frame-deleted ptaA mutant selectively lacked the α-ketoglutaric acid form of pyoverdine, and recombinant PtaA complemented this phenotype. The ptaA / pvdN double mutant produced exclusively the glutamic acid form of pyoverdine. PtaA is homodimeric and contains a pyridoxal phosphate cofactor. Mutation of the active-site lysine abolished PtaA activity and affected folding as well as Tat-dependent transport of the enzyme. In pseudomonads, the occurrence of ptaA correlates with the occurrence of α-ketoglutaric acid forms of pyoverdines. As this enzyme is not restricted to pyoverdine-producing bacteria, its catalysis of periplasmic transaminations is most likely a general tool for specific biosynthetic pathways. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

  4. Characterization of Four New Distinct ω-Transaminases from Pseudomonas putida NBRC 14164 for Kinetic Resolution of Racemic Amines and Amino Alcohols.

    Science.gov (United States)

    Wu, Hua-Lei; Zhang, Jian-Dong; Zhang, Chao-Feng; Fan, Xiao-Jun; Chang, Hong-Hong; Wei, Wen-Long

    2017-03-01

    Four uncharacterized ω-transaminases (ωTAs) from Pseudomonas putida NBRC 14164 have been identified and cloned from the pool of fully sequenced genomes. The genes were functionally expressed in Escherichia coli BL21, and the enzymes were purified and characterized. Four TAs showed highly (S)-selective ωTA activity and converted (S)-α-methylbenzylamine and pyruvate to acetophenone and L-Ala. The maximum activity of cloned enzymes was in the pH range of 8.0-8.5 (Pp36420), 8.5-9.5 (Pp21050), 9.0-9.5 (PpspuC), and 9.5-10.5 (PpbauA), and the optimal temperatures were at 35 °C (Pp36420, Pp21050, and PpspuC) and 50 °C (PpbauA), respectively, with K M of 161.3 mM (Pp21050), 136.7 mM (PpbauA), 398.5 mM (Pp36420), and 130.9 mM (PpspuC) and yielding a catalytic efficiency k cat /K M of 0.015, 0.003, 0.012, and 0.023 mM -1  s -1 . Several racemic amines and amino alcohols were resolved by the cloned ωTAs; perfect conversions (48-50 %) were obtained by at least one enzyme, and the residual substrates were left with 97-99 % ee. Kinetic resolution of racemic phenylglycinol was done with PpspuC in a 100-mL scale. Enaniomeric excess of (S)-phenylglycinol reached 99 % with 45 % isolated yield. The high enantioselectivity and large substrate spectra of the cloned PpTAs showed an attractive potency for biotechnology application in production of chiral amines and amino alcohols.

  5. beta-Alanine elevates dopamine levels in the rat nucleus accumbens: antagonism by strychnine.

    Science.gov (United States)

    Ericson, Mia; Clarke, Rhona B C; Chau, PeiPei; Adermark, Louise; Söderpalm, Bo

    2010-04-01

    Glycine receptors (GlyRs) in the nucleus accumbens (nAc) have recently been suggested to be involved in the reinforcing and dopamine-elevating properties of ethanol via a neuronal circuitry involving the VTA. Apart from ethanol, both glycine and taurine have the ability to modulate dopamine output via GlyRs in the same brain region. In the present study, we wanted to explore whether yet another endogenous ligand for the GlyR, beta-alanine, had similar effects. To this end, we monitored dopamine in the nAc by means of in vivo microdialysis and found that local perfusion of beta-alanine increased dopamine output. In line with previous observations investigating ethanol, glycine and taurine, the competitive GlyR antagonist strychnine completely blocked the dopamine elevation. The present results suggest that beta-alanine has the ability to modulate dopamine levels in the nAc via strychnine-sensitive GlyRs, and are consistent with previous studies suggesting the importance of this receptor for modulating dopamine output.

  6. Volume dose ratios relevant for alanine dosimetry in small, 6 MV photon beams

    International Nuclear Information System (INIS)

    Cronholm, Rickard O.; Andersen, Claus E.; Behrens, Claus F.; Helt-Hansen, Jakob

    2012-01-01

    The overall objective of this work is to establish alanine dosimetry for traceable measurements in clinical radiotherapy beams, in particular for non-reference situations such as small field sizes and composite beam delivery (e.g. intensity modulated radiotherapy and volumetric modulated arc therapy). To this end, we here present the results of a Monte Carlo simulation study with DOSRZnrc that investigated the influence of field and detector size for small 6 MV photon beams. The study focusses on doses averaged over the volume of the detector rather than point doses. The ratio of volume averaged doses to water (D ¯ W ) and alanine (D ¯ det ) was found to be approximately 1.025 for most situations studied, and a constant ratio is likely to be representative for many applications in radiation therapy. However, D ¯ W /D ¯ det was found to be as low as 0.9908 ± 0.0037 in situations where one might expect significant deviations from charged particle equilibrium (i.e. at shallow depths and when the field size was smaller than the range of the secondary electrons). These effects therefore need consideration when finite-size alanine dosimeters are used under such conditions.

  7. Characterisation of L-alanine and glycine absorption across the gut of an ancient vertebrate.

    Science.gov (United States)

    Glover, Chris N; Bucking, Carol; Wood, Chris M

    2011-08-01

    This study utilised an in vitro technique to characterise absorption of two amino acids across the intestinal epithelium of Pacific hagfish, Eptatretus stoutii. Uptake of L-alanine and glycine conformed to Michaelis-Menten kinetics. An uptake affinity (K(m); substrate concentration required to attain a 50% uptake saturation) of 7.0 mM and an uptake capacity (J (max)) of 83 nmol cm(-2) h(-1) were described for L-alanine. The K(m) and J(max) for glycine were 2.2 mM and 11.9 nmol cm(-2) h(-1), respectively. Evidence suggested that the pathways of L-alanine and glycine absorption were shared, and sodium dependent. Further analysis indicated that glycine uptake was independent of luminal pH and proline, but a component of uptake was significantly impaired by 100-fold excesses of threonine or asparagine. The presence of a short-term (24 h) exposure to waterborne glycine, similar in nature to that which may be expected to occur when feeding inside an animal carcass, had no significant impact on gastrointestinal glycine uptake. This may indicate a lack of cross talk between absorptive epithelia. These results are the first published data to describe gastrointestinal uptake of an organic nutrient in the oldest extant vertebrate and may provide potential insight into the evolution of nutrient transport systems.

  8. Determination of isodose curves in Radiotherapy using an Alanine/ESR dosemeter

    International Nuclear Information System (INIS)

    Chen, F.; Baffa, O.; Graeff, C.F.O.

    1998-01-01

    It was studied the possible use of an Alanine/ESR dosemeter in the isodose curves mapping in normal treatments of Radiotherapy. It was manufactured a lot of 150 dosemeters with base in a mixture of D-L Alanine dust (80 %) and paraffin (20 %). Each dosemeter has 4.7 mm diameter and 12 mm length. A group of 100 dosemeters of the lot were arranged inside 50 holes of the slice 25 of the phantom Rando Man. The phantom irradiation was realized in two opposed projections (AP and PA) in Co-60 equipment. A group of 15 dosemeters was take of the same lot for obtaining the calibration curve in a 1-20 Gy range. After irradiation the signal of each dosemeter was measured in an ESR spectrometer operating in the X-band (∼ 9.5 GHz) and the wideness of Alanine ESR spectra central line was correlated with the radiation dose. The wideness dose calibration curve resulted linear with a correlation coefficient 0.9996. The isodose curves obtained show a profile enough similar at comparing with the theoretical curves. (Author)

  9. Alanine dosimetry using a spectrophotometric ferric-xylenol orange complex readout

    International Nuclear Information System (INIS)

    Laere, K. van; Buysse, J.; Berkvens, P.

    1989-01-01

    The spectrophotometric dosimetric method using the indirect oxidation of ferrous ions after dissolution of irradiated DL-and L-alanine has been thoroughly investigated with respect to its composition, read-out procedure and dose-response. Optimal concentration of 0.10 N H 2 SO 4 , 0.2 mM xylenol orange and 0.2 mM Fe 2+ were found, giving an absorption maximum at 547 nm. Standardization of chemical processing procedures allows a reproducibility better than 0.5%. The useful dose range has been extended to 0.03-12 kGy by means of slightly different read-out procedure. The quantitative concept of ''indirect yield'', G id , was introduced for this procedure as a measure of the indirect oxidation capacity of the radicals. It was found to be G id,0 (Fe 3+ ) 7.1 ions/100 eV transferred into the alanine. The spectrophotometric readout combines the highly advantageous use of alanine as a dosemeter with the straightforwardness, accuracy and low costs of the chemical procedure. (author)

  10. 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.

  11. Exogenous alanine and/or glucose plus kanamycin kills antibiotic-resistant bacteria.

    Science.gov (United States)

    Peng, Bo; Su, Yu-Bin; Li, Hui; Han, Yi; Guo, Chang; Tian, Yao-Mei; Peng, Xuan-Xian

    2015-02-03

    Multidrug-resistant bacteria are an increasingly serious threat to human and animal health. However, novel drugs that can manage infections by multidrug-resistant bacteria have proved elusive. Here we show that glucose and alanine abundances are greatly suppressed in kanamycin-resistant Edwardsiella tarda by GC-MS-based metabolomics. Exogenous alanine or glucose restores susceptibility of multidrug-resistant E. tarda to killing by kanamycin, demonstrating an approach to killing multidrug-resistant bacteria. The mechanism underlying this approach is that exogenous glucose or alanine promotes the TCA cycle by substrate activation, which in turn increases production of NADH and proton motive force and stimulates uptake of antibiotic. Similar results are obtained with other Gram-negative bacteria (Vibrio parahaemolyticus, Klebsiella pneumoniae, Pseudomonas aeruginosa) and Gram-positive bacterium (Staphylococcus aureus), and the results are also reproduced in a mouse model for urinary tract infection. This study establishes a functional metabolomics-based strategy to manage infection by antibiotic-resistant bacteria. Copyright © 2015 Elsevier Inc. All rights reserved.

  12. 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.

  13. Prolonged continuous intravenous infusion of the dipeptide L-alanine- L-glutamine significantly increases plasma glutamine and alanine without elevating brain glutamate in patients with severe traumatic brain injury.

    Science.gov (United States)

    Nägeli, Mirjam; Fasshauer, Mario; Sommerfeld, Jutta; Fendel, Angela; Brandi, Giovanna; Stover, John F

    2014-07-02

    Low plasma glutamine levels are associated with worse clinical outcome. Intravenous glutamine infusion dose- dependently increases plasma glutamine levels, thereby correcting hypoglutaminemia. Glutamine may be transformed to glutamate which might limit its application at a higher dose in patients with severe traumatic brain injury (TBI). To date, the optimal glutamine dose required to normalize plasma glutamine levels without increasing plasma and cerebral glutamate has not yet been defined. Changes in plasma and cerebral glutamine, alanine, and glutamate as well as indirect signs of metabolic impairment reflected by increased intracranial pressure (ICP), lactate, lactate-to-pyruvate ratio, electroencephalogram (EEG) activity were determined before, during, and after continuous intravenous infusion of 0.75 g L-alanine-L-glutamine which was given either for 24 hours (group 1, n = 6) or 5 days (group 2, n = 6) in addition to regular enteral nutrition. Lab values including nitrogen balance, urea and ammonia were determined daily. Continuous L-alanine-L-glutamine infusion significantly increased plasma and cerebral glutamine as well as alanine levels, being mostly sustained during the 5 day infusion phase (plasma glutamine: from 295 ± 62 to 500 ± 145 μmol/ l; brain glutamine: from 183 ± 188 to 549 ± 120 μmol/ l; plasma alanine: from 327 ± 91 to 622 ± 182 μmol/ l; brain alanine: from 48 ± 55 to 89 ± 129 μmol/ l; p alanine-L-glutamine infusion (0.75 g/ kg/ d up to 5 days) increased plasma and brain glutamine and alanine levels. This was not associated with elevated glutamate or signs of potential glutamate-mediated cerebral injury. The increased nitrogen load should be considered in patients with renal and hepatic dysfunction. Clinicaltrials.gov NCT02130674. Registered 5 April 2014.

  14. Investigation of complementary methods to the electron paramagnetic resonance for the analysis of radiolysis products formed in alanine by γ irradiation

    International Nuclear Information System (INIS)

    Najosky, Vincent

    1997-01-01

    After a presentation of alanine (a remarkable sensing substance) and of the electron paramagnetic resonance spectroscopy, a discussion of the γ irradiation of alanine and of the radiolytic efficiency, the author reports the design of a glass ampoule aimed at recovering the different gases which may be emitted by irradiated alanine. This ampoule is then used to perform a gas analysis by gas chromatography. The alanine irradiated in the ampoule is then analysed by electron paramagnetic resonance and by mass spectrometry. Using the obtained results, the author determines whether this glass ampoule system provides new information in terms of fundamental radical mechanisms occurring in irradiated alanine [fr

  15. Dosimetry auditing procedure with alanine dosimeters for light ion beam therapy.

    Science.gov (United States)

    Ableitinger, Alexander; Vatnitsky, Stanislav; Herrmann, Rochus; Bassler, Niels; Palmans, Hugo; Sharpe, Peter; Ecker, Swantje; Chaudhri, Naved; Jäkel, Oliver; Georg, Dietmar

    2013-07-01

    In the next few years the number of facilities providing ion beam therapy with scanning beams will increase. An auditing process based on an end-to-end test (including CT imaging, planning and dose delivery) could help new ion therapy centres to validate their entire logistic chain of radiation delivery. An end-to-end procedure was designed and tested in both scanned proton and carbon ion beams, which may also serve as a dosimetric credentialing procedure for clinical trials in the future. The developed procedure is focused only on physical dose delivery and the validation of the biological dose is out of scope of the current work. The audit procedure was based on a homogeneous phantom that mimics the dimension of a head (20 × 20 × 21 cm(3)). The phantom can be loaded either with an ionisation chamber or 20 alanine dosimeters plus 2 radiochromic EBT films. Dose verification aimed at measuring a dose of 10Gy homogeneously delivered to a virtual-target volume of 8 × 8 × 12 cm(3). In order to interpret the readout of the irradiated alanine dosimeters additional Monte Carlo simulations were performed to calculate the energy dependent detector response of the particle fluence in the alanine detector. A pilot run was performed with protons and carbon ions at the Heidelberg Ion Therapy facility (HIT). The mean difference of the absolute physical dose measured with the alanine dosimeters compared with the expected dose from the treatment planning system was -2.4 ± 0.9% (1σ) for protons and -2.2 ± 1.1% (1σ) for carbon ions. The measurements performed with the ionisation chamber indicate this slight underdosage with a dose difference of -1.7% for protons and -1.0% for carbon ions. The profiles measured by radiochromic films showed an acceptable homogeneity of about 3%. Alanine dosimeters are suitable detectors for dosimetry audits in ion beam therapy and the presented end-to-end test is feasible. If further studies show similar results, this dosimetric audit could be

  16. A method to improve application technique in Alanine/ESR dosimetry

    International Nuclear Information System (INIS)

    Choi, Hoon; Ha, Ju Hee; Choi, Won; Lim, Young Khi

    2008-01-01

    Full text: For long-term radiation monitoring to assess the cable aging in harsh condition of nuclear power plant, ESR dosimetry method using alanine dosimeters was already recommended in many technical document and paper. Several ESR dosimetry systems were already produced and used widely, but the actual application of these systems for industrial dosimetry needs careful consideration of error sources in process of dose measurement. The alanine dosimeters were measured by E-scan alanine analyzer system or EMX ESR spectrometer. For the accurate measurement of the dosimeters, we have studied various source of errors. First, this paper discusses sources of inaccuracy related to data processing. To make a meaningful dose assessment, the dosimeter measurements need to be compared with measurements made using certified dosimeters of known dose. This is achieved by performing the routine calibration procedure which creates a calibration curve and corresponding fit coefficients from measurements made with a set of dosimeters with known certified dose. The calibration curves in ESR dosimetry are usually constructed by means of the least-squares technique in its simplest variant. The recommended alternative linearity several replicate measurements of Y at each used X value. Also, measurement is subject to error, so repeat measurements will not be identical. A technical description of a linear calibration is assumed that the dispersion of the measurements is the same for each standard. But in some cases, the standard deviation has to be specified separately for each value of concentration. In this case, it used the WLS (Weighted Least-Squared Regression) method instead of the OLS (Ordinary Least-Squared Regression) method. Second, the precision of the ESR spectrum was showed as a reproducibility test for the two ESR systems. The reproducibility test was performed at an absorbed dose of 1 k Gy, which is at least three orders of magnitude above the background reading of an

  17. β-alanine supplementation improves tactical performance but not cognitive function in combat soldiers.

    Science.gov (United States)

    Hoffman, Jay R; Landau, Geva; Stout, Jeffrey R; Dabora, Matan; Moran, Daniel S; Sharvit, Nurit; Hoffman, Mattan W; Ben Moshe, Yuval; McCormack, William P; Hirschhorn, Gil; Ostfeld, Ishay

    2014-04-10

    There are no known studies that have examined β-alanine supplementation in military personnel. Considering the physiological and potential neurological effects that have been reported during sustained military operations, it appears that β-alanine supplementation may have a potential benefit in maintaining physical and cognitive performance during high-intensity military activity under stressful conditions. The purpose of this study was to examine the effect of 28 days of β-alanine ingestion in military personnel while fatigued on physical and cognitive performance. Twenty soldiers (20.1 ± 0.9 years) from an elite combat unit were randomly assigned to either a β-alanine (BA) or placebo (PL) group. Soldiers were involved in advanced military training, including combat skill development, navigational training, self-defense/hand-to-hand combat and conditioning. All participants performed a 4-km run, 5-countermovement jumps using a linear position transducer, 120-m sprint, a 10-shot shooting protocol with assault rifle, including overcoming a misfire, and a 2-min serial subtraction test to assess cognitive function before (Pre) and after (Post) 28 days of supplementation. The training routine resulted in significant increases in 4-km run time for both groups, but no between group differences were seen (p = 0.597). Peak jump power at Post was greater for BA than PL (p = 0.034), while mean jump power for BA at Post was 10.2% greater (p = 0.139) than PL. BA had a significantly greater (p = 0.012) number of shots on target at Post (8.2 ± 1.0) than PL (6.5 ± 2.1), and their target engagement speed at Post was also significantly faster (p = 0.039). No difference in serial subtraction performance was seen between the groups (p = 0.844). Results of this study indicate that 4-weeks of β-alanine ingestion in young, healthy soldiers did not impact cognitive performance, but did enhance power performance, marksmanship and target

  18. Poli-Β-Alanin Ve Türevlerinin Sentezi, Karakterizasyonu Ve Antibakteriyel Özelliklerinin İncelenmesi

    OpenAIRE

    ÇATIKER, Efkan; ÇİL, Elif; FİLİK, Tahsin

    2018-01-01

    Yakın zamanda naylon-3 (poli-β-alanin) kopolimerleri ve türevlerinin biyoaktivite özellikleri ileilgili çalışmalar artmıştır. Bu çalışmalarda birçok naylon-3 kopolimeri ve türevlerininantibakteriyel, antifungal hatta hücre tutma aktivitesi gösterdiği saptanmıştır. Mevcut çalışmada,poli-β-alanin, poli(α-metil-β-alanin) ve poli(β-metil-β-alanin) polimerleri sentezlenerek yapısalkarakterizasyonu ortaya konmuş ve antibakteriyel aktivitesi incelenmiştir. Sentezlenen üçpolimerin, üç Gram pozitif (B...

  19. 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.

  20. Probing the energy landscape of alanine dipeptide and decalanine using temperature as a tunable parameter in molecular dynamics

    International Nuclear Information System (INIS)

    Chatterjee, A; Bhattacharya, S

    2016-01-01

    We perform several molecular dynamics (MD) calculations of solvated alanine dipeptide and decalanine in vacuum with temperature as a tunable parameter and in the process, generate Markov state models (MSMs) at each temperature. An interesting observation that the kinetic rates appear to obey the Arrhenius rate law allows us to predict the dynamics of alanine dipeptide at 300 K at the microsecond timescales using the nanoseconds long high temperature calculations without actually performing MD simulations at 300 K. We conclude that the energy landscape of alanine dipeptide contains superbasins deeper than k B T and determine the energy barriers associated with the moves from the Arrhenius rate expression. Similar insights regarding the energy landscape associated with folding/unfolding pathways of a deca-alanine molecule are obtained using kinetic rates calculated at different temperatures. (paper)

  1. 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.

  2. Effects of Plyometric Training and Beta-Alanine Supplementation on Maximal-Intensity Exercise and Endurance in Female Soccer Players

    OpenAIRE

    Rosas, Fabi?n; Ram?rez-Campillo, Rodrigo; Mart?nez, Cristian; Caniuqueo, Alexis; Ca?as-Jamet, Rodrigo; McCrudden, Emma; Meylan, Cesar; Moran, Jason; Nakamura, F?bio Y.; Pereira, Lucas A.; Loturco, Irineu; Diaz, Daniela; Izquierdo, Mikel

    2017-01-01

    Abstract Plyometric training and beta-alanine supplementation are common among soccer players, although its combined use had never been tested. Therefore, a randomized, double-blind, placebo-controlled trial was conducted to compare the effects of a plyometric training program, with or without beta-alanine supplementation, on maximal-intensity and endurance performance in female soccer players during an in-season training period. Athletes (23.7 ? 2.4 years) were assigned to either a plyometri...

  3. Inhibiting Inosine Hydrolase and Alanine Racemase to Enhance the Germination of Bacillus anthracis Sterne Spores: Potential Spore Decontamination Strategies

    Science.gov (United States)

    2015-06-19

    2015): << Inhibiting inosine hydrolase and alanine racemase to enhance the germination of Bacillus anthracis Sterne spores: potential spore...display a currently valid OMB control number. 1. REPORT DATE 02 OCT 2015 2. REPORT TYPE N/A 3. DATES COVERED 4. TITLE AND SUBTITLE Inhibiting...inosine hydrolase and alanine racemase to enhance the germination of Bacillus anthracis Sterne spores potential spore decontamination strategies 5a

  4. Human glutamate pyruvate transaminase (GPT): Localization to 8q24.3, cDNA and genomic sequences, and polymorphic sites

    Energy Technology Data Exchange (ETDEWEB)

    Sohocki, M.M.; Sullivan, L.S.; Daiger, S.P. [Univ. of Texas Health Science Center, Houston, TX (United States)] [and others

    1997-03-01

    Two frequent protein variants of glutamate pyruvate transaminase (GPT) (E.C.2.6.1.2) have been used as genetic markers in humans for more than two decades, although chromosomal mapping of the GPT locus in the 1980s produced conflicting results. To resolve this conflict and develop useful DNA markers for this gene, we isolated and characterized cDNA and genomic clones of GPT. We have definitively mapped human GPT to the terminus of 8q using several methods. First, two cosmids shown to contain the GPT sequence were derived from a chromosome 8-specific library. Second, by fluorescence in situ hybridization, we mapped the cosmid containing the human GPT gene to chromosome band 8q24.3. Third, we mapped the rat gpt cDNA to the syntenic region of rat chromosome 7. Finally, PCR primers specific to human GPT amplify sequences contained within a {open_quotes}half-YAC{close_quotes} from the long arm of chromosome 8, that is, a YAC containing the 8q telomere. The human GPT genomic sequence spans 2.7 kb and consists of 11 exons, ranging in size from 79 to 243 bp. The exonic sequence encodes a protein of 495 amino acids that is nearly identical to the previously reported protein sequence of human GPT-1. The two polymorphic GPT isozymes are the result of a nucleotide substitution in codon 14. In addition, a cosmid containing the GPT sequence also contains a previously unmapped, polymorphic microsatellite sequence, D8S421. The cloned GPT gene and associated polymorphisms will be useful for linkage and physical mapping of disease loci that map to the terminus of 8q, including atypical vitelliform macular dystrophy (VMD1) and epidermolysis bullosa simplex, type Ogna (EBS1). In addition, this will be a useful system for characterizing the telomeric region of 8q. Finally, determination of the molecular basis of the GPT isozyme variants will permit PCR-based detection of this world-wide polymorphism. 22 refs., 3 figs.

  5. Status report of the ESR/alanine project of the IAEA Dosimetry Laboratory

    International Nuclear Information System (INIS)

    Girzikowsky, R.

    1990-01-01

    The main tasks of the Dosimetry Laboratory of the IAEA are in the field of therapy-level dosimetry. Other dose ranges, i.e. protection-level standardization and calibration, are partly covered, too. On high-dose level the laboratory has been offering a Fricke-dosimetry service to Member States on request since 1965. In 1985 the Dosimetry Section has initiated an International Dose Assurance Service (IDAS). This service is an important part of the Agency's high-dose standardization programme and is based on Electron-Spin-Resonance (ESR) analysis of radiation-induced free radicals in alanine. This ESR/alanine dosimetry system was initially developed for high-dose application by GSF/Munich as the Agency's outside contractor. Although efforts were undertaken to equip the Agency Laboratory with an ESR analyzer since 1984, the purchase of an adequate unit was postponed until the end of 1988. From the date of establishment of the IDAS programme until today, the handling of all technical aspects, i.e. dosimeter production and evaluation was and is carried out by GSP/Munich under IAEA contract. As mentioned above, the IAEA Dosimetry Laboratory is in possession of its own ESR analyzer since December 1988. It was then installed in May 1989. Only one staff member of the laboratory was assigned to this project. He made himself acquainted with the analyzer, the measuring technique, and the application of alanine as dosimeter material. The ESR spectrometric analyzer is a CW ESR type ESP 300 manufactured by Bruker GmbH/FRG. It consists of a 9''/2,7 kW magnet, an X-band microwave bridge, a field regulator unit, a signal channel unit and a data system ESP 1620 based on the 68020 CPU. The probe material selected for measurement of concentration of radiation-induced free radicals is L-Alanine. This amino acid is a suitable material for transfer dosimeters due to its properties, i.e. wide sensitivity range (10 Gy to 100 kGy), energy independence for high-energy photons and electron

  6. Free Energy Landscapes of Alanine Oligopeptides in Rigid-Body and Hybrid Water Models.

    Science.gov (United States)

    Nayar, Divya; Chakravarty, Charusita

    2015-08-27

    Replica exchange molecular dynamics is used to study the effect of different rigid-body (mTIP3P, TIP4P, SPC/E) and hybrid (H1.56, H3.00) water models on the conformational free energy landscape of the alanine oligopeptides (acAnme and acA5nme), in conjunction with the CHARMM22 force field. The free energy landscape is mapped out as a function of the Ramachandran angles. In addition, various secondary structure metrics, solvation shell properties, and the number of peptide-solvent hydrogen bonds are monitored. Alanine dipeptide is found to have similar free energy landscapes in different solvent models, an insensitivity which may be due to the absence of possibilities for forming i-(i + 4) or i-(i + 3) intrapeptide hydrogen bonds. The pentapeptide, acA5nme, where there are three intrapeptide backbone hydrogen bonds, shows a conformational free energy landscape with a much greater degree of sensitivity to the choice of solvent model, though the three rigid-body water models differ only quantitatively. The pentapeptide prefers nonhelical, non-native PPII and β-sheet populations as the solvent is changed from SPC/E to the less tetrahedral liquid (H1.56) to an LJ-like liquid (H3.00). The pentapeptide conformational order metrics indicate a preference for open, solvent-exposed, non-native structures in hybrid solvent models at all temperatures of study. The possible correlations between the properties of solvent models and secondary structure preferences of alanine oligopeptides are discussed, and the competition between intrapeptide, peptide-solvent, and solvent-solvent hydrogen bonding is shown to be crucial in the relative free energies of different conformers.

  7. 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

  8. Amino Acid Isotopic Trophic Enrichment in Mesozooplankton: Is Alanine a Better Predictor of Protistan Grazer Steps?

    Science.gov (United States)

    Decima, M.; Landry, M. R.; Bradley, C. J.; Fogel, M. L.

    2016-02-01

    Food-web studies within marine environments are increasingly reliant upon results from compound-specific isotope analysis of amino acids (CSIA-AA). The approach is advantageous because it allows consumer trophic positions to be estimated without sampling the dynamic primary producers. The baseline signal in the source AA phenylalanine is preserved, and a constant enrichment in glutamic acid at each trophic step is assumed, regardless of consumer type or diet. However, a number of recent studies challenge the assumption of universal and invariant isotopic fractionation of glutamic acid for all trophic levels, as well as its specific applicability to the main grazers in the ocean: the protistan microzooplankton. We present results from both laboratory and field studies that further explore this issue. Experiments include six 2-stage chemostats, using two different microzooplankton-phytoplankton pairs and one copepod-phytoplankton pair, and one 3-stage experiment using a copepod-microzooplankton-phytoplankton chain. We confirm previous observations of negligible fractionation of glutamic acid in protistan consumers when nutrients are limiting. In contrast, a consistent trophic enrichment effect was observed for alanine, with increasing δ15N values by trophic level for both metazoan and protistan consumers. A re-analysis of published CSIA-AA data of zooplankton species show that an index using alanine and phenylalanine gives trophic level estimates closer to expected given current understanding of the linkages within microbial food webs. Our results examine the details of isotopic fractionation of alanine within defined food chains and generally support its potential use as a trophic level indicator that includes the protistan contribution to mesozooplankton diet.

  9. Alanine scan of core positions in ubiquitin reveals links between dynamics, stability, and function.

    Science.gov (United States)

    Lee, Shirley Y; Pullen, Lester; Virgil, Daniel J; Castañeda, Carlos A; Abeykoon, Dulith; Bolon, Daniel N A; Fushman, David

    2014-04-03

    Mutations at solvent-inaccessible core positions in proteins can impact function through many biophysical mechanisms including alterations to thermodynamic stability and protein dynamics. As these properties of proteins are difficult to investigate, the impacts of core mutations on protein function are poorly understood for most systems. Here, we determined the effects of alanine mutations at all 15 core positions in ubiquitin on function in yeast. The majority (13 of 15) of alanine substitutions supported yeast growth as the sole ubiquitin. Both the two null mutants (I30A and L43A) were less stable to temperature-induced unfolding in vitro than wild type (WT) but were well folded at physiological temperatures. Heteronuclear NMR studies indicated that the L43A mutation reduces temperature stability while retaining a ground-state structure similar to WT. This structure enables L43A to bind to common ubiquitin receptors in vitro. Many of the core alanine ubiquitin mutants, including one of the null variants (I30A), exhibited an increased accumulation of high-molecular-weight species, suggesting that these mutants caused a defect in the processing of ubiquitin-substrate conjugates. In contrast, L43A exhibited a unique accumulation pattern with reduced levels of high-molecular-weight species and undetectable levels of free ubiquitin. When conjugation to other proteins was blocked, L43A ubiquitin accumulated as free ubiquitin in yeast. Based on these findings, we speculate that ubiquitin's stability to unfolding may be required for efficient recycling during proteasome-mediated substrate degradation. Copyright © 2013 Elsevier Ltd. All rights reserved.

  10. Dose verification and calibration of the Cyberknife by EPR/alanine dosimetry

    Energy Technology Data Exchange (ETDEWEB)

    Garcia, Tristan, E-mail: tristan.garcia@cea.fr [CEA, LIST, Laboratoire National Henri Becquerel, 91191 Gif-sur-Yvette Cedex (France); Lacornerie, Thomas [Service de Physique Medicale, Centre Oscar-Lambret, 3, rue Frederic-Combemale, 59020 Lille (France); Popoff, Romain; Lourenco, Valerie; Bordy, Jean-Marc [CEA, LIST, Laboratoire National Henri Becquerel, 91191 Gif-sur-Yvette Cedex (France)

    2011-09-15

    Standard calibration protocols in radiotherapy (AAPM TG-51 and IAEA TRS 398) recommend a 10 x 10 cm{sup 2} irradiation field at a distance of 100 cm. However, the specific geometry of the Cyberknife machine enables only circular radiation beams with a maximum diameter of 6 cm field size at a distance of 80 cm from the source which prevents from a direct use of these protocols. Thereby, specific calibration and verification protocols are required. Within this framework, the Laboratoire National Henri Becquerel and the Centre Oscar-Lambret decided to study two points of interest: i) the feasibility of the calibration of the Cyberknife beam; ii) the verification of the delivered dose during a treatment with a very small diameter beam using Electron Paramagnetic Resonance (EPR)/alanine dosimetry. Alanine dosimeters are suitable for this work thanks to their small size, their integrating capability, and their low energy dependence as a function of both dose rate and beam energy in the radiotherapy dose range. The results of the dose rate calibration using EPR/alanine are very good, their deviations compared to the dose rate measured in the Cyberknife reference conditions are small (about 0.5%). The doses delivered during a treatment, measured within a 2% standard uncertainty, showed less than 4% deviation from the doses calculated by the treatment planning system. So, a rather good agreement is found between the measured and theoretically delivered doses. Nevertheless, the smaller the collimator diameter is, the larger the deviation is. This problem could be due to the evaluation of the output factor correction which is difficult to achieve for such small fields or to the Treatment Planning System algorithm itself and in particular the profile modelling.

  11. Small-Field Dosimetry in A 6 MV Photon Beam Using Alanine and Liquid Ionisation Chamber

    DEFF Research Database (Denmark)

    Zimmermann, S.; Riis, H. L.; Hjelm-Hansen, M.

    2012-01-01

    Purpose/Objective: Dosimetry of small field sizes in MV photon beams is an increasingly important subject, and a generally accepted guideline for clinical measurements is still lacking. The present comparative study was carried out to further investigate the use of alanine and the PTW microLion...... of each field and depth. This dose maximum was measured for each field using a Scanditronix Wellhöfer photon field diode. The same measurements were carried out using a liquid ionchamber, PTW microLion, irradiated by 500 MU. The output of the accelerator was controlled by a PTW semiflex ion chamber...

  12. Evaluation of alanine as a reference dosimeter for therapy level dose comparisons in megavoltage electron beams

    International Nuclear Information System (INIS)

    McEwen, Malcolm; Sharpe, Peter; Voros, Sandor

    2015-01-01

    When comparing absorbed dose standards from different laboratories (e.g. National Measurement Institutes, NMIs, for Key or Supplementary comparisons) it is rarely possible to carry out a direct comparison of primary standard instruments, and therefore some form of transfer detector is required. Historically, air-filled, unsealed ionization chambers have been used because of the long history of using these instruments, very good stability over many years, and ease of transport. However, the use of ion chambers for therapy-level comparisons is not without its problems. Findings from recent investigations suggest that ion chambers are prone to non-random variations, they are not completely robust to standard courier practices, and failure at any step in a comparison can render all measurements potentially useless. An alternative approach is to identify a transfer system that is insensitive to some of these concerns - effectively a dosimeter that is inexpensive, simple to use, robust, but with sufficient precision and of a size relevant to the disseminated quantity in question. The alanine dosimetry system has been successfully used in a number of situations as an audit dosimeter and therefore the purpose of this investigation was to determine whether alanine could also be used as the transfer detector for dosimetric comparisons, which require a lower value for the measurement uncertainty. A measurement protocol was developed for comparing primary standards of absorbed dose to water in high-energy electron beams using alanine pellets irradiated in a water-equivalent plastic phantom. A trial comparison has been carried out between three NMIs and has indicated that alanine is a suitable alternative to ion chambers, with the system used achieving a precision of 0.1%. Although the focus of the evaluation was on the performance of the dosimeter, the comparison results are encouraging, showing agreement at the level of the combined uncertainties (∼0.6%). Based on this

  13. Inhibition study of alanine aminotransferase enzyme using sequential online capillary electrophoresis analysis.

    Science.gov (United States)

    Liu, Lina; Chen, Yuanfang; Yang, Li

    2014-12-15

    We report the study of several inhibitors on alanine aminotransferase (ALT) enzyme using sequential online capillary electrophoresis (CE) assay. Using metal ions (Na(+) and Mg(2+)) as example inhibitors, we show that evolution of the ALT inhibition reaction can be achieved by automatically and simultaneously monitoring the substrate consumption and product formation as a function of reaction time. The inhibition mechanism and kinetic constants of ALT inhibition with succinic acid and two traditional Chinese medicines were derived from the sequential online CE assay. Our study could provide valuable information about the inhibition reactions of ALT enzyme. Copyright © 2014 Elsevier Inc. All rights reserved.

  14. Theoretical investigation of radical species formed from L-α-alanine under gamma-irradiation

    International Nuclear Information System (INIS)

    Simion, C.

    2008-01-01

    Gamma-irradiated L-α-alanine used in EPR-coupled dosimetry has a complex EPR spectrum at room temperature. Changing the temperature or other conditions of the irradiated samples leads to varied EPR spectrum, i.e., some components disappear and/or new ones are formed. We used both molecular mechanics (MM+) and semiempirical (AM1) methods to perform a theoretical investigation of the seven radical species that have been experimentally detected. We established their order of priority in the given simulation conditions (at 0 K, in vacuo). The formation stages advanced for these long-lived radical species were characterized by a theoretical determination of the reaction enthalpies. (author)

  15. Aspartate Aminotransferase (AST/GOT) and Alanine Aminotransferase (ALT/GPT) Detection Techniques

    Science.gov (United States)

    Huang, Xing-Jiu; Choi, Yang-Kyu; Im, Hyung-Soon; Yarimaga, Oktay; Yoon, Euisik; Kim, Hak-Sung

    2006-01-01

    The levels of aspartate aminotransferase (AST/GOT) and alanine aminotransferase (ALT/GPT) in serum can help people diagnose body tissues especially the heart and the liver are injured or not. This article provides a comprehensive review of research activities that concentrate on AST/GOT and ALT/GPT detection techniques due to their clinical importance. The detection techniques include colorimetric, spectrophotometric, chemiluminescence, chromatography, fluorescence and UV absorbance, radiochemical, and electrochemical techniques. We devote the most attention on experimental principle. In some methods a few representative devices and important conclusions are presented.

  16. A comparative study of MP2, B3LYP, RHF and SCC-DFTB force fields in predicting the vibrational spectra of N-acetyl-L-alanine-N'-methyl amide: VA and VCD spectra

    DEFF Research Database (Denmark)

    Bohr, Henrik; Jalkanen, Karl J.; Elstner, M.

    1999-01-01

    Recently we have looked for spectroscopic probes for secondary structural elements in the vibrational spectra of N-acetyl-L-alanine N'-methyl amide (NALANMA), L-alanine (LA), N-acetyl-L-alanyl-L-alanine N'-methyl amide (NALALANMA) and L-alanyl-L-alanine (LALA). Our goal has been to identify spect...

  17. 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)

  18. Verification of the pure alanine in PMMA tube dosimeter applicability for dosimetry of radiotherapy photon beams: a feasibility study.

    Science.gov (United States)

    Al-Karmi, Anan M; Ayaz, Ali Asghar H; Al-Enezi, Mamdouh S; Abdel-Rahman, Wamied; Dwaikat, Nidal

    2015-09-01

    Alanine dosimeters in the form of pure alanine powder in PMMA plastic tubes were investigated for dosimetry in a clinical application. Electron paramagnetic resonance (EPR) spectroscopy was used to measure absorbed radiation doses by detection of signals from radicals generated in irradiated alanine. The measurements were performed for low-dose ranges typical for single-fraction doses often used in external photon beam radiotherapy. First, the dosimeters were irradiated in a solid water phantom to establish calibration curves in the dose range from 0.3 to 3 Gy for 6 and 18 MV X-ray beams from a clinical linear accelerator. Next, the dosimeters were placed at various locations in an anthropomorphic pelvic phantom to measure the dose delivery of a conventional four-field box technique treatment plan to the pelvis. Finally, the doses measured with alanine dosimeters were compared against the doses calculated with a commercial treatment planning system (TPS). The results showed that the alanine dosimeters have a highly sensitive dose response with good linearity and no energy dependence in the dose range and photon beams used in this work. Also, a fairly good agreement was found between the in-phantom dose measurements with alanine dosimeters and the TPS dose calculations. The mean value of the ratios of measured to calculated dose values was found to be near unity. The measured points in the in-field region passed dose-difference acceptance criterion of 3% and those in the penumbral region passed distance-to-agreement acceptance criterion of 3 mm. These findings suggest that the pure alanine powder in PMMA tube dosimeter is a suitable option for dosimetry of radiotherapy photon beams.

  19. 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.

  20. The effects of beta alanine plus creatine administration on performance during repeated bouts of supramaximal exercise in sedentary men.

    Science.gov (United States)

    Okudan, N; Belviranli, M; Pepe, H; Gökbel, H

    2015-11-01

    The aim of this study was to investigate the effects of beta alanine and/or creatine supplementation on performance during repeated bouts of supramaximal exercise in sedentary men. Forty-four untrained healthy men (aged 20-22 years, weight: 68-72 kg, height: 174-178 cm) participated in the present study. After performing the Wingate Test (WAnT) for three times in the baseline exercise session, the subjects were assigned to one of four treatment groups randomly: 1) placebo (P; 10 g maltodextrose); 2) creatine (Cr; 5 g creatine plus 5 g maltodextrose); 3) beta-alanine (β-ALA; 1,6 g beta alanine plus 8,4 g maltodextrose); and 4) beta-alanine plus creatine (β-ALA+Cr; 1,6 g beta alanine plus 5 g creatine plus 3,4 g maltodextrose). Participants were given the supplements orally twice a day for 22 consecutive days, then four times a day for the following 6 days. After 28 days, the second exercise session was applied during which peak power (PP) and mean power (MP) were measured and fatigue index (FI) was calculated. PP and MP decreased and FI increased in all groups during exercise before and after the treatment. During the postsupplementation session PP2 and PP3 increased in creatine supplemented group (from 642.7±148.6 to 825.1±205.2 in PP2 and from 522.9±117.5 to 683.0±148.0 in PP3, respectively). However, MP increased in β-ALA+Cr during the postsupplementation compared to presupplementation in all exercise sessions (from 586.2±55.4 to 620.6±49.6 in MP1, from 418.1±37.2 to 478.3±30.3 in MP2 and from 362.0±41.3 to 399.1±3 in MP3, respectively). FI did not change with beta alanine and beta alanine plus creatine supplementation during the postsupplementation exercise session. Beta-alanine and beta alanine plus creatine supplementations have strong performance enhancing effect by increasing mean power and delaying fatigue Index during the repeated WAnT.

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

    Directory of Open Access Journals (Sweden)

    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.

  2. Enhanced poly(3-hydroxypropionate production via β-alanine pathway in recombinant Escherichia coli.

    Directory of Open Access Journals (Sweden)

    Stephen Tamekou Lacmata

    Full Text Available Poly(3-hydroxypropionate (P3HP is a thermoplastic with great compostability and biocompatibility, and can be produced through several biosynthetic pathways, in which the glycerol pathway achieved the highest P3HP production. However, exogenous supply of vitamin B12 was required to maintain the activity of glycerol dehydratase, resulting in high production cost. To avoid the addition of VB12, we have previously constructed a P3HP biosynthetic route with β-alanine as intermediate, and the present study aimed to improve the P3HP production of this pathway. L-aspartate decarboxylase PanD was found to be the rate-limiting enzyme in the β-alanine pathway firstly. To improve the pathway efficiency, PanD was screened from four different sources (Escherichia coli, Bacillus subtilis, Pseudomonas fluorescens, and Corynebacterium glutamicum. And PanD from C. glutamicum was found to have the highest activity, the P3HP production was improved in flask cultivation with this enzyme. To further improve the production, the host strain was screened and the culture condition was optimized. Under optimal conditions, production and content of P3HP reached to 10.2 g/L and 39.1% (wt/wt [cell dry weight] in an aerobic fed-batch fermentation. To date, this is the highest P3HP production without VB12.

  3. A single glycine-alanine exchange directs ligand specificity of the elephant progestin receptor.

    Directory of Open Access Journals (Sweden)

    Michael Wierer

    Full Text Available The primary gestagen of elephants is 5α-dihydroprogesterone (DHP, which is unlike all other mammals studied until now. The level of DHP in elephants equals that of progesterone in other mammals, and elephants are able to bind DHP with similar affinity to progesterone indicating a unique ligand-binding specificity of the elephant progestin receptor (PR. Using site-directed mutagenesis in combination with in vitro binding studies we here report that this change in specificity is due to a single glycine to alanine exchange at position 722 (G722A of PR, which specifically increases DHP affinity while not affecting binding of progesterone. By conducting molecular dynamics simulations comparing human and elephant PR ligand-binding domains (LBD, we observed that the alanine methyl group at position 722 is able to push the DHP A-ring into a position similar to progesterone. In the human PR, the DHP A-ring position is twisted towards helix 3 of PR thereby disturbing the hydrogen bond pattern around the C3-keto group, resulting in a lower binding affinity. Furthermore, we observed that the elephant PR ligand-binding pocket is more rigid than the human analogue, which probably explains the higher affinity towards both progesterone and DHP. Interestingly, the G722A substitution is not elephant-specific, rather it is also present in five independent lineages of mammalian evolution, suggesting a special role of the substitution for the development of distinct mammalian gestagen systems.

  4. Direct detection of second harmonic and its use in alanine/EPR dosimetry

    International Nuclear Information System (INIS)

    Chen, F.; Guzman, C.S.; Graeff, C.F.O.; Baffa, O.

    2001-01-01

    In this work, the possible use of the second harmonic EPR signal from irradiated alanine for low radiation dose (∼1 Gy) was explored, aiming applications to HDR brachytherapy and teletherapy. The second harmonic signal was directly detected after overmodulation. A batch of DL-alanine/paraffin small cylindrical pellets was made. A VARIAN E-4 X-Band EPR spectrometer with optimized operation parameters like microwave power and modulation amplitude to obtain a signal with the highest amplitude was used. The modulation frequency and modulation amplitude were 100 kHz and 1.25 mT (to overmodulate the signal) respectively. The second harmonic signal was directly detected at twice the modulation frequency. One group of dosimeters was irradiated with a 192 Ir brachytherapy source and the other in a 10 MeV X-rays linear accelerator, both group at a dose range: 0.5 - 15 Gy. The second harmonic signal showed better resolution than the first harmonic one making possible a more easy localization of the signal. Moreover, for both types of radiation, the dose-response curve showed a good linear behavior for the dose range indicated. (author)

  5. First Calibrations of Alanine and Radio-Photo-Luminescence Dosemeters to a Hadronic Radiation Environment

    CERN Document Server

    Fürstner, Markus; Floret, Idelette; Forkel-Wirth, Doris; Mayer, Sabine; Menzel, Hans Gregor; Vincke, Helmut H

    2005-01-01

    Alanine and Radio-Photo-Luminescence (RPL) dosimeters are used to monitor radiation doses occurring inside the tunnels of all CERN accelerators including the Large Hadron Collider (LHC). They are placed close to radiation sensitive machine components like cables or insulation of magnet coils to predict their remaining lifetime. The dosimeters are exposed to mixed high-energy radiation fields. However, up to now both dosimeter types are calibrated to 60Co-photons only. In order to study the response of RPL and alanine dosimeters to mixed particle fields like those occurring at CERN's accelerators, an irradiation campaign at the CERN-EC High-Energy Reference field Facility (CERF-field) was performed. Moreover, the dosimeters were first time calibrated to a proton radiation field of a constant momentum of 24 GeV/c. In addition to the experiment FLUKA Monte Carlo simulations were carried out, which provide information concerning the energy deposition and the radiation field at the dosimeter locations.

  6. In vivo dose evaluation during gynaecological radiotherapy using L-alanine/ESR dosimetry

    International Nuclear Information System (INIS)

    Burg Rech, Amanda; Baffa, Oswaldo; Barbi, Gustavo Lazzaro; Almeida Ventura, Luiz Henrique; Silva Guimaraes, Flavio; Oliveira, Harley Francisco

    2014-01-01

    The dose delivered by in vivo 3-D external beam radiation therapy (EBRT) was verified with L-alanine/electron spin resonance (ESR) dosimetry for patients diagnosed with gynaecological cancer. Measurements were performed with an X-band ESR spectrometer. Dosemeters were positioned inside the vaginal cavity with the assistance of an apparatus specially designed for this study. Previous phantom studies were performed using the same conditions as in the in vivo treatment. Four patients participated in this study during 20-irradiation sessions, giving 220 dosemeters to be analysed. The doses were determined with the treatment planning system, providing dose confirmation. The phantom study resulted in a deviation between -2.5 and 2.1 %, and for the in vivo study a deviation between -9.2 and 14.2 % was observed. In all cases, the use of alanine with ESR was effective for dose assessment, yielding results consistent with the values set forth in the International Commission on Radiation Units and Measurements (ICRU) reports. (authors)

  7. Role of tRNAPro in pretransfer editing of alanine by prolyl-tRNA synthetase

    Directory of Open Access Journals (Sweden)

    Boyarshin K. S.

    2013-09-01

    Full Text Available Aim. To characterize the process of tRNA-dependent pretransfer edi- ting of alanine by prolyl-tRNA synthetase of bacteria Enterococcus faecalis (ProRSEf. Methods. Velocity of the editing processes in vitro was determined by ATP hydrolysis by ProRSEf. Pretransfer and posttransfer editing were experimentally separated by site-directed mutagenesis. Results. tRNA-dependent pretransfer editing is characterized by three-fold larger velocity then tRNA-independent editing. Effectivity of the process depends on the presence of 2'-hydroxyle group of A76 tRNAPro. In the absence of tRNAPro selective release of alanyl-AMP occurs simultaneously with tRNA-independent pretransfer editing. Released alanyl-AMP can be re-bound and hydrolyzed. Conclusions. tRNA-dependent pretransfer editing of alanine by ProRSEf is the catalytic mechanism, mediated by 2'-hydroxyl group of A76 tRNAPro. In the absence of tRNAPro tRNA-independent pretransfer editing and selective release of alanyl-AMP occur.

  8. Determinants of Alanine Dipeptide Conformational Equilibria on Graphene and Hydroxylated Derivatives.

    Science.gov (United States)

    Poblete, Horacio; Miranda-Carvajal, Ingrid; Comer, Jeffrey

    2017-04-20

    Understanding the interaction of carbon nanomaterials with proteins is essential for determining the potential effects of these materials on health and in the design of biotechnology based on them. Here we leverage explicit-solvent molecular simulation and multidimensional free-energy calculations to investigate how adsorption to carbon nanomaterial surfaces affects the conformational equilibrium of alanine dipeptide, a widely used model of protein backbone structure. We find that the two most favorable structures of alanine dipeptide on graphene (or large carbon nanotubes) correspond to the two amide linkages lying in the same plane, flat against the surface, rather than the nonplanar α-helix-like and β-sheet-like conformations that predominate in aqueous solution. On graphenic surfaces, the latter conformations are metastable and most often correspond to amide-π stacking of the N-terminal amide. The calculations highlight the key role of amide-π interactions in determining the conformational equilibrium. Lesser but significant contributions from hydrogen bonding to the high density interfacial water layer or to the hydroxy groups of hydroxylated graphene also define the most favorable conformations. This work should yield insight on the influence of carbon nanotubes, graphene, and their functionalized derivatives on protein structure.

  9. 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)

  10. 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.

  11. Adsorption characteristics of 14C-labeled alanine, aspartic acid and adenosine triphosphate by metal-chelating resins

    International Nuclear Information System (INIS)

    Ishiyama, Toshio; Matsunami, Tadao; Shibata, Setsuko; Honda, Yoshihide.

    1987-01-01

    (1) Adsorption properties of 14 C-alanine, 14 C-ATP (adenosine triphosphate) and 14 C-aspartic acid on the metal-chelating resins were determined and found that the Cu(II)-Chelex 100 and Fe(III)-Unicellex UR10, Fe(III)-Chelex 100 chelating resins were highly effective for the adsorption of 14 C-alanine and 14 C-ATP, respectively. (2) Desorption rate of 14 C-ATP from the Fe(III)-Unicellex UR10 and Fe(III)-Chelex 100 resins was somewhat higher than the case of 14 C-alanine, probably because the coordination bonds of Cu-alanine might be stronger than those of Fe-ATP. Thus, 14 C-labeled organic compounds such as 14 C-alanine and 14 C-ATP of a low activity concentration (3.7 mBq/ml) (1 x 10 -7 μCi/ml) in aqueous solution may be measured with liquid scintillation counter after pre-concentration by use of the Fe(III)- and Cu(II)-chelating resin columns. (author)

  12. 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)

  13. 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.

  14. Establishing a synthetic pathway for high-level production of 3-hydroxypropionic acid in Saccharomyces cerevisiae via β-alanine

    DEFF Research Database (Denmark)

    Borodina, Irina; Kildegaard, Kanchana Rueksomtawin; Jensen, Niels Bjerg

    2015-01-01

    . With the objective of developing Saccharomyces cerevisiae as an efficient cell factory for highlevel production of 3HP, we identified the ß-alanine biosynthetic route as the most economically attractive according to the metabolic modeling. We engineered and optimized a synthetic pathway for de novo biosynthesis of ß......-alanine and its subsequent conversion into 3HP using a novel ß-alanine-pyruvate aminotransferase discovered in Bacillus cereus. The final strain produced 3HP at a titer of 13.7±0.3 g・L-1 with a 0.14±0.0 C-mol・C-mol-1 yield on glucose in 80 hours in controlled fed-batch fermentation in mineral medium at pH 5...

  15. Beta-alanine supplementation improves jumping power and affects severe-intensity performance in professional alpine skiers.

    Science.gov (United States)

    Gross, Micah; Bieri, Kathrin; Hoppeler, Hans; Norman, Barbara; Vogt, Michael

    2014-12-01

    Supplementation with beta-alanine may have positive effects on severe-intensity, intermittent, and isometric strength-endurance performance. These could be advantageous for competitive alpine skiers, whose races last 45 to 150 s, require metabolic power above the aerobic maximum, and involve isometric muscle work. Further, beta-alanine supplementation affects the muscle force-frequency relationship, which could influence explosiveness. We explored the effects of beta-alanine on explosive jump performance, severe exercise energy metabolism, and severe-intensity ski-like performance. Nine male elite alpine skiers consumed 4.8 g/d beta-alanine or placebo for 5 weeks in a double-blind fashion. Before and after, they performed countermovement jumps (CMJ), a 90-s cycling bout at 110% VO2max (CLT), and a maximal 90-s box jump test (BJ90). Beta-alanine improved maximal (+7 ± 3%, d = 0.9) and mean CMJ power (+7 ± 2%, d = 0.7), tended to reduce oxygen deficit (-3 ± 8%, p = .06) and lactate accumulation (-12 ± 31%) and enhance aerobic energy contribution (+1.3 ± 2.9%, p = .07) in the CLT, and improved performance in the last third of BJ90 (+7 ± 4%, p = .02). These effects were not observed with placebo. Beta-alanine supplementation improved explosive and repeated jump performance in elite alpine skiers. Enhanced muscle contractility could possibly explain improved explosive and repeated jump performance. Increased aerobic energy production could possibly help explain repeated jump performance as well.

  16. Overexpression of Genes Encoding Glycolytic Enzymes in Corynebacterium glutamicum Enhances Glucose Metabolism and Alanine Production under Oxygen Deprivation Conditions

    Science.gov (United States)

    Yamamoto, Shogo; Gunji, Wataru; Suzuki, Hiroaki; Toda, Hiroshi; Suda, Masako; Jojima, Toru; Inui, Masayuki

    2012-01-01

    We previously reported that Corynebacterium glutamicum strain ΔldhAΔppc+alaD+gapA, overexpressing glyceraldehyde-3-phosphate dehydrogenase-encoding gapA, shows significantly improved glucose consumption and alanine formation under oxygen deprivation conditions (T. Jojima, M. Fujii, E. Mori, M. Inui, and H. Yukawa, Appl. Microbiol. Biotechnol. 87:159–165, 2010). In this study, we employ stepwise overexpression and chromosomal integration of a total of four genes encoding glycolytic enzymes (herein referred to as glycolytic genes) to demonstrate further successive improvements in C. glutamicum glucose metabolism under oxygen deprivation. In addition to gapA, overexpressing pyruvate kinase-encoding pyk and phosphofructokinase-encoding pfk enabled strain GLY2/pCRD500 to realize respective 13% and 20% improved rates of glucose consumption and alanine formation compared to GLY1/pCRD500. Subsequent overexpression of glucose-6-phosphate isomerase-encoding gpi in strain GLY3/pCRD500 further improved its glucose metabolism. Notably, both alanine productivity and yield increased after each overexpression step. After 48 h of incubation, GLY3/pCRD500 produced 2,430 mM alanine at a yield of 91.8%. This was 6.4-fold higher productivity than that of the wild-type strain. Intracellular metabolite analysis showed that gapA overexpression led to a decreased concentration of metabolites upstream of glyceraldehyde-3-phosphate dehydrogenase, suggesting that the overexpression resolved a bottleneck in glycolysis. Changing ratios of the extracellular metabolites by overexpression of glycolytic genes resulted in reduction of the intracellular NADH/NAD+ ratio, which also plays an important role on the improvement of glucose consumption. Enhanced alanine dehydrogenase activity using a high-copy-number plasmid further accelerated the overall alanine productivity. Increase in glycolytic enzyme activities is a promising approach to make drastic progress in growth-arrested bioprocesses. PMID

  17. Medical reference dosimetry using EPR measurements of alanine: Development of an improved method for clinical dose levels

    International Nuclear Information System (INIS)

    Helt-Hansen, Jakob; Andersen, Claus Erik; Rosendal, Flemming; Kofoed, Inger Matilde

    2009-01-01

    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 signal fading, non-destructive measurement and small dosimeter size. Material and Methods. A Bruker EMX-micro EPR spectrometer with a rectangular cavity and a measurement time of two minutes per dosimeter was used for reading of irradiated alanine dosimeters. Under these conditions a new algorithm based on scaling of known spectra was developed to extract the alanine signal. Results. The dose accuracy, including calibration uncertainty, is less than 2% (k=1) above 4 Gy (n=4). The measurement uncertainty is fairly constant in absolute terms (∼30 mGy) and the relative uncertainty therefore rises for dose measurements below 4 Gy. Typical reproducibility is <1% (k=1) above 10 Gy and <2% between 4 and 10 Gy. Below 4 Gy the uncertainty is higher. A depth dose curve measurement was performed in a solid-water phantom irradiated to a dose of 20 Gy at the maximum dose point (dmax) in 6 and 18 MV photon beams. The typical difference between the dose measured with alanine in solid water and the dose measured with an ion chamber in a water tank was about 1%. A difference of 2% between 6 and 18 MV was found, possibly due to non-water equivalence of the applied phantom. Discussion. Compared to previously published 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

  18. A structure-based design approach for the identification of novel inhibitors: application to an alanine racemase

    Science.gov (United States)

    Mustata, Gabriela Iurcu; Briggs, James M.

    2002-12-01

    We report a new structure-based strategy for the identification of novel inhibitors. This approach has been applied to Bacillus stearothermophilus alanine racemase (AlaR), an enzyme implicated in the biosynthesis of the bacterial cell wall. The enzyme catalyzes the racemization of l- and d-alanine using pyridoxal 5'-phosphate (PLP) as a cofactor. The restriction of AlaR to bacteria and some fungi and the absolute requirement for d-alanine in peptidoglycan biosynthesis make alanine racemase a suitable target for drug design. Unfortunately, known inhibitors of alanine racemase are not specific and inhibit the activity of other PLP-dependent enzymes, leading to neurological and other side effects. This article describes the development of a receptor-based pharmacophore model for AlaR, taking into account receptor flexibility (i.e. a `dynamic' pharmacophore model). In order to accomplish this, molecular dynamics (MD) simulations were performed on the full AlaR dimer from Bacillus stearothermophilus (PDB entry, 1sft) with a d-alanine molecule in one active site and the non-covalent inhibitor, propionate, in the second active site of this homodimer. The basic strategy followed in this study was to utilize conformations of the protein obtained during MD simulations to generate a dynamic pharmacophore model using the property mapping capability of the LigBuilder program. Compounds from the Available Chemicals Directory that fit the pharmacophore model were identified and have been submitted for experimental testing. The approach described here can be used as a valuable tool for the design of novel inhibitors of other biomolecular targets.

  19. 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.

  20. The Biological Properties and Potential Interacting Proteins of d-Alanyl-d-alanine Ligase A from Mycobacterium tuberculosis

    Directory of Open Access Journals (Sweden)

    Shufeng Yang

    2018-02-01

    Full Text Available (1 Background: d-alanine-d-alanine ligase (DdlA, an effective target for drug development to combat against Mycobacterium tuberculosis (Mtb, which threatens human health globally, supplies a substrate of d-alanyl-d-alanine for peptidoglycan crosslinking by catalyzing the dimerization of two d-alanines. To obtain a better understanding of DdlA profiles and develop a colorimetric assay for high-throughput inhibitor screening, we focused on explicating and characterizing Tb-DdlA. (2 Methods and Results: Rv2981c (ddlA was expressed in Escherichia coli, and the purified Tb-DdlA was identified using (anti-polyhistidine antibody followed by DdlA activity confirmation by measuring the released orthophosphate via colorimetric assay and the yielded d-alanyl-d-alanine through high performance thin layer chromatography (HP-TLC. The kinetic assays on Tb-DdlA indicated that Tb-DdlA exhibited a higher affinity to ATP (KmATP: 50.327 ± 4.652 μmol/L than alanine (KmAla: 1.011 ± 0.094 mmol/L. A colorimetric assay for Tb-DdlA activity was developed for high-throughput screening of DdlA inhibitors in this study. In addition, we presented an analysis on Tb-DdlA interaction partners by pull-down assay and MS/MS. Eight putative interaction partners of Tb-DdlA were identified. (3 Conclusions: Our dataset provided a valuable resource for exploring Tb-DdlA biology, and developed an easy colorimetric assay for screening of Tb-DdlA inhibitors.

  1. Effect of radiation on the in vivo degradation of DL-alanine-containing copolypeptides

    International Nuclear Information System (INIS)

    Asano, Masaharu; Yoshida, Masaru; Kaetsu, Isao; Oya, Masanao; Imai, Kyoichi; Mashimo, Tooru; Yuasa, Hisako; Yamanaka, Hidetoshi; Suzuki, Keiji.

    1985-01-01

    Some random DL-alanine-containing copolymers, such as DL-alanine [Ala] and β-ethyl-L-aspartate [Asp(OEt)] and DL-alanine [Ala] and γ-ethyl-L-glutamate [Glu(OEt)] were prepared according to N-carboxy α-amino acid anhydride (NCA) methods. They were then melted by heating under a pressure of 150 kg/cm 2 to obtain the columnar matrices (1.6 mm in diameter) which have high density and rigidity. The 100 % in vivo degradation of melt-pressed copolymer matrices were observed in compositions more than 50 mol% Ala for [AlaAsp(OEt)]sub(n) and 75 mol% Ala for [AlaGlu(OEt)]sub(n). After a melt-pressed [AlaAsp(OEt)]sub(n) (50 mol% Ala) was irradiated for 3 h at a dose rate of 1 x 10 6 rad/h at temperatures of -196, -78, 0, 30, and 60 0 C, in vacuo with γ-rays from a 60 Co source, the in vivo degradations when they were implanted subcutaneously in the back of rats during the first 3 weeks period were 33.6 %, 29.5, 18.9, 52.5, and 22.4 %, respectively. Thus the in vivo degradation of the above matrix has the maximum value at around 30 0 C. Such a tendency was observed also in a [AlaGlu(OEt)]sub(n) matrix. The viscosity (etasub(sp)/c) of a melt-pressed [AlaAsp(OEt)]sub(n) (50 mol% Ala) irradiated at a temperature of 0 0 C rapidly decreased with an increase in the irradiation dose (1 x 10 5 rad to 1 x 10 7 rad: 0.50 dl/g to 0.13 dl/g), though it gradually decreased at 30 0 C (1 x 10 5 rad to 1 x 10 7 rad: 0.53 dl/g to 0.33 dl/g). In this case, the in vivo degradation of melt-pressed matrix irradiated at 0 0 C decreased with an increase in the irradiation dose. The data of amino acid analysis showed that the digestion of Ala moiety in a [AlaAsp(OEt)]sub(n) (50 mol% Ala) irradiated at 30 0 C is slightly faster than that at 0 0 C. (J.P.N.)

  2. Characterization of the functional epitope on the urokinase receptor. Complete alanine scanning mutagenesis supplemented by chemical cross-linking

    DEFF Research Database (Denmark)

    Gårdsvoll, Henrik; Gilquin, Bernard; Le Du, Marie Hélène

    2006-01-01

    a comprehensive alanine scanning mutagenesis of uPAR combined with low resolution distance constraints defined within the complex using chemical cross-linkers as molecular rulers. The kinetic rate constants for the interaction between pro-uPA and 244 purified uPAR mutants with single-site replacements were...... determined by surface plasmon resonance. This complete alanine scanning of uPAR highlighted the involvement of 20 surface-exposed side chains in this interaction. Mutations causing delta deltaG > or = 1 kcal/mol for the uPA interaction are all located within or at the rim of the central cavity uniquely...

  3. SU-E-T-608: Perturbation Corrections for Alanine Dosimeters in Different Phantom Materials in High-Energy Photon Beams

    International Nuclear Information System (INIS)

    Voigts-Rhetz, P von; Czarnecki, D; Anton, M; Zink, K

    2015-01-01

    Purpose: Alanine dosimeters are often used for in-vivo dosimetry purposes in radiation therapy. In a Monte Carlo study the influence of 20 different surrounding/phantom materials for alanine dosimeters was investigated. The investigations were performed in high-energy photon beams, covering the whole range from 60 Co up to 25 MV-X. The aim of the study is the introduction of a perturbation correction k env for alanine dosimeters accounting for the environmental material. Methods: The influence of different surrounding materials on the response of alanine dosimeters was investigated with Monte Carlo simulations using the EGSnrc code. The photon source was adapted with BEAMnrc to a 60 Co unit and an Elekta (E nom =6, 10, 25 MV-X) linear accelerator. Different tissue-equivalent materials ranging from cortical bone to lung were investigated. In addition to available phantom materials, some material compositions were taken and scaled to different electron densities. The depth of the alanine detectors within the different phantom materials corresponds to 5 cm depth in water, i.e. the depth is scaled according to the electron density (n e /n e,w ) of the corresponding phantom material. The dose was scored within the detector volume once for an alanine/paraffin mixture and once for a liquid water voxel. The relative response, the ratio of the absorbed dose to alanine to the absorbed dose to water, was calculated and compared to the corresponding ratio under reference conditions. Results: For each beam quality the relative response r and the correction factor for the environment kenv was calculated. k env =0.9991+0.0049 *((n e /n e,w )−0.7659) 3 Conclusion: A perturbation correction factor k env accounting for the phantom environment has been introduced. The response of the alanine dosimeter can be considered independent of the surrounding material for relative electron densities (n e /n e,w ) between 1 and 1.4. For denser materials such as bone or much less dense

  4. High-temperature Raman study of L-alanine, L-threonine and taurine crystals related to thermal decomposition

    International Nuclear Information System (INIS)

    Cavaignac, A.L.O.; Lima, R.J.C.; Façanha Filho, P.F.; Moreno, A.J.D.; Freire, P.T.C.

    2016-01-01

    In this work high-temperature Raman spectra are used to compare temperature dependence of the lattice mode wavenumber of L-alanine, L-threonine and taurine crystals. Anharmonic effects observed are associated with intermolecular N-H· · ·O hydrogen bond that plays an important role in thermal decomposition process of these materials. Short and strong hydrogen bonds in L-alanine crystal were associated with anharmonic effects in lattice modes leading to low thermal stability compared to taurine crystals. Connection between thermal decomposition process and anharmonic effects is furnished for the first time.

  5. High-temperature Raman study of L-alanine, L-threonine and taurine crystals related to thermal decomposition

    Science.gov (United States)

    Cavaignac, A. L. O.; Lima, R. J. C.; Façanha Filho, P. F.; Moreno, A. J. D.; Freire, P. T. C.

    2016-03-01

    In this work high-temperature Raman spectra are used to compare temperature dependence of the lattice mode wavenumber of L-alanine, L-threonine and taurine crystals. Anharmonic effects observed are associated with intermolecular N-H· · ·O hydrogen bond that plays an important role in thermal decomposition process of these materials. Short and strong hydrogen bonds in L-alanine crystal were associated with anharmonic effects in lattice modes leading to low thermal stability compared to taurine crystals. Connection between thermal decomposition process and anharmonic effects is furnished for the first time.

  6. The effect of irradiation temperatures between ambient and 80 deg. C on the response of alanine dosimeters

    DEFF Research Database (Denmark)

    Sharpe, P.H.G.; Miller, Arne; Sephton, J.P.

    2009-01-01

    dosimeters at temperatures up to 80 °C and doses up to 70 kGy. Data have been obtained for both 60Co and electron beam irradiations and the effect of temperature on the stability of the radiation-induced signal has also been investigated. At temperatures above 50 °C the irradiation temperature coefficient......Published data on the effect of irradiation temperature on the response of alanine dosimeters does not extend to the temperatures that may be experienced in high-dose industrial irradiations, particularly in the case of electron beams. We describe here results of the irradiation of alanine...

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

    Directory of Open Access Journals (Sweden)

    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.

  8. Assessment of an alanine EPR dosimetry technique with enhanced precision and accuracy

    International Nuclear Information System (INIS)

    Hayes, Robert B.; Haskell, E.H.; Wieser, Albrecht; Romanyukha, Alexander A.; Hardy, Byron L.; Barrus, Jeffrey K.

    2000-01-01

    Dose reconstruction in the course of a series of blind tests demonstrated that an accuracy of 10 mGy for low doses and 1% for high doses can be achieved using EPR spectroscopy. This was accomplished using a combination of methodologies including polynomial filtration of the EPR spectrum, dosimeter rotation during scanning, use of an EPR standard fixed into the resonator and subtraction of all nonradiogenic signals. Doses were reconstructed over the range of 0.01-1000 Gy using this compound spectral EPR analysis. This EPR technique, being equally applicable to fractionated doses (such as those delivered during multiple radiotherapy treatments), was verified to exhibit dose reciprocity. Irradiated alanine dosimeters which were stored exhibited compound spectral EPR signal fading of ca 3% over 9 months. All error estimates given in this paper are given at the 1 standard deviation level and unless otherwise specified do not account for uncertainties in source calibration

  9. Thiophenyl-substituted triazolyl-thione L-alanine: asymmetric synthesis, aggregation and biological properties.

    Science.gov (United States)

    Saghyan, Ashot S; Simonyan, Hayarpi M; Petrosyan, Satenik G; Geolchanyan, Arpine V; Roviello, Giovanni N; Musumeci, Domenica; Roviello, Valentina

    2014-10-01

    In this work, we report the asymmetric synthesis and characterization of an artificial amino acid based on triazolyl-thione L-alanine, which was modified with a thiophenyl-substituted moiety, as well as in vitro studies of its nucleic acid-binding ability. We found, by dynamic light scattering studies, that the synthetic amino acid was able to form supramolecular aggregates having a hydrodynamic diameter higher than 200 nm. Furthermore, we demonstrated, by UV and CD experiments, that the heteroaromatic amino acid, whose enzymatic stability was demonstrated by HPLC analysis also after 24 h of incubation in human serum, was able to bind a RNA complex, which is a feature of biomedical interest in view of innovative antiviral strategies based on modulation of RNA-RNA molecular recognition.

  10. Discovery of an L-alanine ester prodrug of the Hsp90 inhibitor, MPC-3100.

    Science.gov (United States)

    Kim, Se-Ho; Tangallapally, Rajendra; Kim, In Chul; Trovato, Richard; Parker, Daniel; Patton, J Scott; Reeves, Leslie; Bradford, Chad; Wettstein, Daniel; Baichwal, Vijay; Papac, Damon; Bajji, Ashok; Carlson, Robert; Yager, Kraig M

    2015-11-15

    Various types of Hsp90 inhibitors have been and continue to undergo clinical investigation. One development candidate is the purine-based, synthetic Hsp90 inhibitor 1 (MPC-3100), which successfully completed a phase I clinical study. However, further clinical development of 1 was hindered by poor solubility and consequent formulation issues and promoted development of a more water soluble prodrug. Towards this end, numerous pro-moieties were explored in vitro and in vivo. These studies resulted in identification of L-alanine ester mesylate, 2i (MPC-0767), which exhibited improved aqueous solubility, adequate chemical stability, and rapid bioconversion without the need for solubilizing excipients. Based on improved physical characteristics and favorable PK and PD profiles, 2i mesylate was selected for further development. A convergent, scalable, chromatography-free synthesis for 2i mesylate was developed to support further clinical evaluation. Copyright © 2015 Elsevier Ltd. All rights reserved.

  11. Assessment of an alanine EPR dosimetry technique with enhanced precision and accuracy

    CERN Document Server

    Hayes, R B; Wieser, A; Romanyukha, A A; Hardy, B L; Barrus, J K

    2000-01-01

    Dose reconstruction in the course of a series of blind tests demonstrated that an accuracy of 10 mGy for low doses and 1% for high doses can be achieved using EPR spectroscopy. This was accomplished using a combination of methodologies including polynomial filtration of the EPR spectrum, dosimeter rotation during scanning, use of an EPR standard fixed into the resonator and subtraction of all nonradiogenic signals. Doses were reconstructed over the range of 0.01-1000 Gy using this compound spectral EPR analysis. This EPR technique, being equally applicable to fractionated doses (such as those delivered during multiple radiotherapy treatments), was verified to exhibit dose reciprocity. Irradiated alanine dosimeters which were stored exhibited compound spectral EPR signal fading of ca 3% over 9 months. All error estimates given in this paper are given at the 1 standard deviation level and unless otherwise specified do not account for uncertainties in source calibration.

  12. 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.

  13. 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.

  14. Growth and characterization of organometallic L-alanine cadmium chloride single crystal by slow evaporation technique

    International Nuclear Information System (INIS)

    Bright, K.C.; Freeda, T.H.

    2010-01-01

    Single crystals of L-alanine cadmium chloride (LACC), an organometallic nonlinear optical material, have been grown by the slow evaporation technique. The grown crystals were subjected to various characterization techniques, such as single crystal and powder XRD, FTIR, UV-vis and TGA-DTA. The mechanical properties of the crystals show that this material belongs to the category of hard materials. Second harmonic generation was confirmed by the Kurtz and Perry powder technique. Electrical parameters, such as dielectric constant, dielectric loss, ac and dc conductivity and their corresponding activation energies have been studied. The low dielectric constant and dielectric loss suggest that this material is a good candidate for micro-electronic applications.

  15. Abdominal obesity validates the association between elevated alanine aminotransferase and newly diagnosed diabetes mellitus.

    Science.gov (United States)

    Yueh, Chen-Yu; Yang, Yao-Hsu; Sung, Yi-Ting; Lee, Li-Wen

    2014-01-01

    To examine how elevated alanine aminotransferase (ALT) could be associated with newly diagnosed diabetes mellitus. We conducted a cross-sectional analysis on a mass health examination. The odds ratios (ORs) for diabetes mellitus and newly diagnosed diabetes mellitus were compared between people with and without abdominal obesity, together with and without elevated ALT levels. 5499 people were included in this study. Two hundred fifty two (4.6%) fulfilled the diagnosis of diabetes mellitus with 178 (3.2%) undiagnosed before. Metabolic syndrome was vigorously associated with diabetes mellitus and newly diagnosed diabetes mellitus (12.4% vs. 1.4% and 9.0% vs. 0.9%), but elevated ALT alone was not. However, coexisting with obesity, elevated ALTs were robustly associated with diabetes mellitus and newly diagnosed diabetes mellitus. For the incidence of newly diagnosed diabetes mellitus, in comparison to non-obese people with normal ALT (1.7%, OR = 1), obese people especially with elevated ALT levels had significantly higher ORs (obese with ALT ≤ 40 U/L: 4.7%, OR 1.73, 95% CI 1.08-2.77, P 0.023; ALT 41-80 U/L: 6.8%, OR 2.06, 95% CI 1.20-3.55, P 0.009; ALT 81-120 U/L: 8.8%, OR 3.07, 95% CI 1.38-6.84, P 0.006; ALT > 120 U/L: 18.2%, OR 7.44, 95% CI 3.04-18.18, P obesity validates the association between elevated alanine aminotransferase and diabetes mellitus and newly diagnosed diabetes mellitus. People with abdominal obesity, especially with coexisting elevated ALT levels should be screened for undiagnosed diabetes mellitus.

  16. Alanine screening mutagenesis establishes the critical inactivating damage of irradiated E. coli lactose repressor.

    Science.gov (United States)

    Goffinont, Stephane; Villette, Sandrine; Spotheim-Maurizot, Melanie

    2012-06-01

    The function of the E. coli lactose operon requires the binding of lactose repressor to operator DNA. We have previously shown that γ rradiation destabilizes the repressor-operator complex because the repressor loses its DNA-binding ability. It was suggested that the observed oxidation of the four tyrosines (Y7, Y12, Y17, Y47) and the concomitant structural changes of the irradiated DNA-binding domains (headpieces) could be responsible for the inactivation. To pinpoint the tyrosine whose oxidation has the strongest effect, four headpieces containing the product of tyrosine oxidation, 3,4-dihydroxyphenylalanine (DOPA), were simulated by molecular dynamics. We have observed that replacing Y47 by DOPA triggers the largest change of structure and stability of the headpiece and have concluded that Y47 oxidation is the greatest contributor to the decrease of repressor binding to DNA. To experimentally verify this conclusion, we applied the alanine screening mutagenesis approach. Tetrameric mutated repressors bearing an alanine instead of each one of the tyrosines were prepared and their binding to operator DNA was checked. Their binding ability is quite similar to that of the wild-type repressor, except for the Y47A mutant whose binding is strongly reduced. Circular dichroism determinations revealed small reductions of the proportion of α helices and of the melting temperature for Y7A, Y12A and Y17A headpieces, but much larger ones were revealed for Y47A headpiece. These results established the critical role of Y47 oxidation in modifying the structure and stability of the headpiece, and in reduction of the binding ability of the whole lactose repressor.

  17. 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)

  18. Effect of lysine to alanine mutations on the phosphate activation and BPTES inhibition of glutaminase.

    Science.gov (United States)

    McDonald, Charles J; Acheff, Eric; Kennedy, Ryan; Taylor, Lynn; Curthoys, Norman P

    2015-09-01

    The GLS1 gene encodes a mitochondrial glutaminase that is highly expressed in brain, kidney, small intestine and many transformed cells. Recent studies have identified multiple lysine residues in glutaminase that are sites of N-acetylation. Interestingly, these sites are located within either a loop segment that regulates access of glutamine to the active site or the dimer:dimer interface that participates in the phosphate-dependent oligomerization and activation of the enzyme. These two segments also contain the binding sites for bis-2[5-phenylacetamido-1,2,4-thiadiazol-2-yl]ethylsulfide (BPTES), a highly specific and potent uncompetitive inhibitor of this glutaminase. BPTES is also the lead compound for development of novel cancer chemotherapeutic agents. To provide a preliminary assessment of the potential effects of N-acetylation, the corresponding lysine to alanine mutations were constructed in the hGACΔ1 plasmid. The wild type and mutated proteins were purified by Ni(+)-affinity chromatography and their phosphate activation and BPTES inhibition profiles were analyzed. Two of the alanine substitutions in the loop segment (K311A and K328A) and the one in the dimer:dimer interface (K396A) form enzymes that require greater concentrations of phosphate to produce half-maximal activation and exhibit greater sensitivity to BPTES inhibition. By contrast, the K320A mutation results in a glutaminase that exhibits near maximal activity in the absence of phosphate and is not inhibited by BPTES. Thus, lysine N-acetylation may contribute to the acute regulation of glutaminase activity in various tissues and alter the efficacy of BPTES-type inhibitors. Copyright © 2014 Elsevier Ltd. All rights reserved.

  19. Alanine and proline content modulate global sensitivity to discrete perturbations in disordered proteins.

    Science.gov (United States)

    Perez, Romel B; Tischer, Alexander; Auton, Matthew; Whitten, Steven T

    2014-12-01

    Molecular transduction of biological signals is understood primarily in terms of the cooperative structural transitions of protein macromolecules, providing a mechanism through which discrete local structure perturbations affect global macromolecular properties. The recognition that proteins lacking tertiary stability, commonly referred to as intrinsically disordered proteins (IDPs), mediate key signaling pathways suggests that protein structures without cooperative intramolecular interactions may also have the ability to couple local and global structure changes. Presented here are results from experiments that measured and tested the ability of disordered proteins to couple local changes in structure to global changes in structure. Using the intrinsically disordered N-terminal region of the p53 protein as an experimental model, a set of proline (PRO) and alanine (ALA) to glycine (GLY) substitution variants were designed to modulate backbone conformational propensities without introducing non-native intramolecular interactions. The hydrodynamic radius (R(h)) was used to monitor changes in global structure. Circular dichroism spectroscopy showed that the GLY substitutions decreased polyproline II (PP(II)) propensities relative to the wild type, as expected, and fluorescence methods indicated that substitution-induced changes in R(h) were not associated with folding. The experiments showed that changes in local PP(II) structure cause changes in R(h) that are variable and that depend on the intrinsic chain propensities of PRO and ALA residues, demonstrating a mechanism for coupling local and global structure changes. Molecular simulations that model our results were used to extend the analysis to other proteins and illustrate the generality of the observed PRO and alanine effects on the structures of IDPs. © 2014 Wiley Periodicals, Inc.

  20. 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.

  1. Oral administration of D-alanine in monkeys robustly increases plasma and cerebrospinal fluid levels but experimental D-amino acid oxidase inhibitors had minimal effect.

    Science.gov (United States)

    Rojas, Camilo; Alt, Jesse; Ator, Nancy A; Wilmoth, Heather; Rais, Rana; Hin, Niyada; DeVivo, Michael; Popiolek, Michael; Tsukamoto, Takashi; Slusher, Barbara S

    2016-09-01

    Hypofunction of the N-methyl-d-aspartate (NMDA) receptor is thought to exacerbate psychosis in patients diagnosed with schizophrenia. Consistent with this hypothesis, D-alanine, a co-agonist at the glycine site of the NMDA receptor, was shown to improve positive and cognitive symptoms when used as add-on therapy for schizophrenia treatment. However, D-alanine had to be administered at high doses (~7 g) to observe clinical effects. One possible reason for the high dose is that D-alanine could be undergoing oxidation by D-amino acid oxidase (DAAO) before it reaches the brain. If this is the case, the dose could be reduced by co-administration of D-alanine with a DAAO inhibitor (DAAOi). Early studies with rodents showed that co-administration of D-alanine with 5-chloro-benzo[d]isoxazol-3-ol (CBIO), a prototype DAAOi, significantly enhanced the levels of extracellular D-alanine in the frontal cortex compared with D-alanine alone. Further, the use of CBIO reduced the dose of D-alanine needed to attenuate prepulse inhibition deficits induced by dizocilpine. The objective of the work reported herein was to confirm the hypothesis that DAAO inhibition can enhance D-alanine exposure in a species closer to humans: non-human primates. We report that while oral D-alanine administration to baboons (10 mg/kg) enhanced D-alanine plasma and CSF levels over 20-fold versus endogenous levels, addition of experimental DAAOi to the regimen exhibited a 2.2-fold enhancement in plasma and no measurable effect on CSF levels. The results provide caution regarding the utility of DAAO inhibition to increase D-amino acid levels as treatment for patients with schizophrenia. © The Author(s) 2016.

  2. Alanine aminotransferase and the 6-year risk of the metabolic syndrome in Caucasian men and women : The Hoorn Study

    NARCIS (Netherlands)

    Schindhelm, R. K.; Dekker, J. M.; Nijpels, G.; Stehouwer, C. D A; Bouter, L. M.; Heine, R. J.; Diamant, M.

    Aims: To study the association between alanine aminotransferase (ALT) and the 6-year risk of the metabolic syndrome in a population-based study in Caucasian men and women. Methods: The association of ALT with the 6-year risk of the metabolic syndrome in 1097 subjects, aged 50-75 years, was assessed

  3. 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

  4. Dosimetry in non-homogeneous media with alanine/EPR mini dosemeters and simulation with PENELOPE Monte Carlo code

    International Nuclear Information System (INIS)

    Vega Ramirez, J.L.; Chen, F.; Nicolucci, P.; Baffa, O.

    2009-01-01

    The dosimetric system of L-alanine mini dosimeter and K-Band EPR spectrometer was tested for the dosimetry in non-homogeneous media through the determination of the Percentage Depth Dose (PDD) curve for a small radiation field. The alanine mini dosimeters were produced by mechanical pressure of a mixture of L-alanine (95%) and PVA (5%) to nominal dimensions of 1 mm diameter and 3 mm length and 3 - 4 mg. For detecting the EPR signal of the mini dosimeters irradiated to 25 Gy, a K-Band (24 GHz) spectrometer was used. The dosimeters were irradiated in a 60 Co radiotherapy unit using 80 cm source skin distance and field sizes of 2.5 x 2.5 cm 2 . The inhomogeneous phantom consisted of acrylic and cork sheets of 30 x 30 x 1 cm 3 ; six cork sheets were sandwiched between five and nine acrylic sheets, which were placed at the top and bottom regions respectively. PDD curves with radiographic film and PENELOPE simulation were also determined. The PDD results for alanine mini dosimeters agreed better than 5.9% with film and PENELOPE. (author)

  5. Hydrophobic radical influence on structure and vibration spectra of zwitter-ionic forms of glycine and alanine in condensed state

    International Nuclear Information System (INIS)

    Ten, G.N.; Kadrov, D.M.; Baranov, V.I.

    2014-01-01

    Structure and vibrational spectra of the zwitter-ionic forms of glycine and alanine in water solution and solid state have been calculated in the B3LYP/6-311++G(d,p) approximation. The environment influence has been taken into account by two methods: the self-consistent reaction field (SCRF) method and one of modeling the glycine and alanine complexes with molecules of water. The structure, energy and spectral properties have been determined which allow establishing an influence of the hydrophobic radical on the glycine and alanine ability to form the hydrogen bonds. It is shown by comparison with experiment that for the calculation of vibrational (IR and Raman) spectra of the zwitter-ionic forms of glycine and alanine in the condensed states they must be surrounded with three molecules of water, one of which is located between the N + H 3 and COO - ionic groups. The value of energy necessary to form the Ala complexes with water compared to Gly ones is 56.47 and 12.55 kcal/mol higher in the case of the complex formation with 1and 3 molecules of water, respectively, located between bipolar groups. (authors)

  6. Crystallization and preliminary X-ray analysis of beta-alanine synthase from the yeast Saccharomyces kluyveri

    DEFF Research Database (Denmark)

    Dobritzsch, D.; Gojkovic, Zoran; Andersen, Birgit

    2003-01-01

    In eukaryotes and some bacteria, the third step of reductive pyrimidine catabolism is catalyzed by beta-alanine synthase (EC 3.5.1.6). Crystals of the recombinant enzyme from the yeast Saccharomyces kluyveri were obtained using sodium citrate as a precipitant. The crystals belong to space group P2...

  7. High alanine aminotransferase is associated with decreased hepatic insulin sensitivity and predicts the development of type 2 diabetes

    DEFF Research Database (Denmark)

    Vozarova, Barbora; Stefan, Norbert; Lindsay, Robert S

    2002-01-01

    It has been proposed that liver dysfunction may contribute to the development of type 2 diabetes. The aim of the present study was to examine whether elevated hepatic enzymes (alanine aminotransferase [ALT], aspartate aminotransferase [AST], or gamma -glutamyltranspeptidase [GGT]) are associated...

  8. Global Transcriptional and Physiological Responses of Saccharomyces cerevisiae to Ammonium, L-Alanine, or L-Glutamine Limitation

    DEFF Research Database (Denmark)

    Usaite, Renata; Patil, Kiran Raosaheb; Grotkjær, Thomas

    2006-01-01

    The yeast Saccharomyces cerevisiae encounters a range of nitrogen sources at various concentrations in its environment. The impact of these two parameters on transcription and metabolism was studied by growing S. cerevisiae in chemostat cultures with L-glutamine, L-alanine, or L-ammonium in limit......The yeast Saccharomyces cerevisiae encounters a range of nitrogen sources at various concentrations in its environment. The impact of these two parameters on transcription and metabolism was studied by growing S. cerevisiae in chemostat cultures with L-glutamine, L-alanine, or L......-ammonium in limitation and by growing cells in an excess of ammonium. Cells grown in L-alanine-limited cultures had higher biomass yield per nitrogen mole (19%) than those from ammonium-limited cultures. Whole-genome transcript profiles were analyzed with a genome-scalle metabolic model that suggested increased anabolic...... activity in L-alanine-limited cells. The changes in these cells were found to be focused around pyruvate, acetyl coenzyme A, glyoxylate, and alpha-ketoglutarate via increased levels of ALT1, DAL7, PYC1, GDH2, and ADH5 and decreased levels of GDH3, CIT2, and ACS1 transcripts. The transcript profiles were...

  9. Metadynamics As a Tool for Mapping the Conformational and Free-Energy Space of Peptides - The Alanine Dipeptide Case Study

    Czech Academy of Sciences Publication Activity Database

    Vymětal, Jiří; Vondrášek, Jiří

    2010-01-01

    Roč. 114, č. 16 (2010), s. 5632-5642 ISSN 1520-6106 R&D Projects: GA MŠk LC512 Institutional research plan: CEZ:AV0Z40550506 Keywords : metadynamics * alanine dipeptide * explicit water models Subject RIV: CF - Physical ; Theoretical Chemistry Impact factor: 3.603, year: 2010

  10. Increased alanine aminotransferase levels and associated characteristics among newly diagnosed type 2 diabetes patients: Results from the DD2 study

    DEFF Research Database (Denmark)

    Mor, Anil; Thomsen, Reimar W.; Rungby, Jørgen

    Objectives: Elevated levels of serum alanine aminotransferase (ALAT) have been linked with non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), insulin resistance and the metabolic syndrome in type 2 diabetes (T2D) patients. We examined ALAT levels in newly diagnosed T2D...

  11. Conformational flexibility of L-alanine zwitterion determines shapes of Raman and Raman optical activity spectral bands

    Czech Academy of Sciences Publication Activity Database

    Kapitán, Josef; Baumruk, V.; Kopecký ml., V.; Bouř, Petr

    2006-01-01

    Roč. 110, č. 14 (2006), s. 4689-4696 ISSN 1089-5639 R&D Projects: GA ČR(CZ) GA203/06/0420 Institutional research plan: CEZ:AV0Z40550506 Keywords : Raman optical activity * molecular flexibility * alanine Subject RIV: CF - Physical ; Theoretical Chemistry Impact factor: 3.047, year: 2006

  12. Effect of tolbutamide on 14C-sodium bicarbonate and 14C-alanine metabolism in isolated rat hepatocytes

    International Nuclear Information System (INIS)

    Kunjathoor, V.V.; Ye, Y.; Pillai, U.A.; Ferguson, P.W.; Medon, P.J.

    1990-01-01

    Tolbutamide (TOLB) is a sulfonylurea commonly used in the treatment of noninsulin-dependent diabetes mellitus. Studies have shown that TOLB affects gluconeogenesis and glycolysis from various substrates in the liver. Specifically, TOLB inhibits gluconeogenesis from lactate in a dose-dependent manner. In order to further clarify tolbutamide's mechanism of action, its effect on the incorporation of 14 C from NaH 14 CO 3 and 14 C-alanine into glucose, lactate or pyruvate in the presence of lactate was measured. Rat hepatocytes were incubated with lactate (2.0 mM) with or without TOLB (1.0 mM) in the presence of NaH 14 CO 3 or 14 C-alanine. TOLB inhibited the incorporation of C 14 from NaHCO 3 and alanine into glucose by 55 and 56%, respectively. TOLB did not alter the incorporation of C 14 from NaHCO 3 into lactate or pyruvate. TOLB did not affect the incorporation of 14 C from alanine into lactate but produced a pooling of 14 C as pyruvate. The authors data support studies demonstrating the TOLB produces its actions, in part, by increasing the concentration of fructose-2,6-bisphosphate and inhibiting pyruvate carboxylase

  13. Risk of alanine transferase (ALT) elevation in patients with rheumatoid arthritis treated with methotrexate in a DAS-steered strategy

    NARCIS (Netherlands)

    Dirven, L.; Klarenbeek, N.B.; van den Broek, M.; Groenendael, J.H.L.M.; de Sonnaville, P.B.J.; Kerstens, P.J.S.M.; Huizinga, T.W.J.; Dijkmans, B.A.C.; Lems, W.F.; Allaart, C.F.

    2013-01-01

    Objective: To determine incidence of increased levels of alanine transferase (ALT) >2× upper limit of normal (ULN) in patients receiving methotrexate (MTX), treated according to a dynamic strategy, and to identify predictors of ALT of >2× ULN. Methods: Data of 508 recent-onset rheumatoid arthritis

  14. 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

  15. Amino acid metabolism of Astacus leptodactylus Esch.—III. Studies on the biosynthesis of α- and β-alanine from aspartate

    NARCIS (Netherlands)

    Marrewijk, W.J.A. van; Zandee, D.I.

    1975-01-01

    1. 1. Six hours after injection of 1- or 4-14C-aspartate into the crayfish Astacus leptodactylus almost all radioactivity incorporated was found in the amino acids. 2. 2. From both precursors only the amino acids α-alanine and glutamic acid were labelled. The biosynthesis of α-alanine from

  16. β-alanine supplementation to improve exercise capacity and performance: a systematic review and meta-analysis.

    Science.gov (United States)

    Saunders, Bryan; Elliott-Sale, Kirsty; Artioli, Guilherme G; Swinton, Paul A; Dolan, Eimear; Roschel, Hamilton; Sale, Craig; Gualano, Bruno

    2017-04-01

    To conduct a systematic review and meta-analysis of the evidence on the effects of β-alanine supplementation on exercise capacity and performance. This study was designed in accordance with PRISMA guidelines. A 3-level mixed effects model was employed to model effect sizes and account for dependencies within data. 3 databases (PubMed, Google Scholar, Web of Science) were searched using a number of terms ('β-alanine' and 'Beta-alanine' combined with 'supplementation', 'exercise', 'training', 'athlete', 'performance' and 'carnosine'). Inclusion/exclusion criteria limited articles to double-blinded, placebo-controlled studies investigating the effects of β-alanine supplementation on an exercise measure. All healthy participant populations were considered, while supplementation protocols were restricted to chronic ingestion. Cross-over designs were excluded due to the long washout period for skeletal muscle carnosine following supplementation. A single outcome measure was extracted for each exercise protocol and converted to effect sizes for meta-analyses. 40 individual studies employing 65 different exercise protocols and totalling 70 exercise measures in 1461 participants were included in the analyses. A significant overall effect size of 0.18 (95% CI 0.08 to 0.28) was shown. Meta-regression demonstrated that exercise duration significantly (p=0.004) moderated effect sizes. Subgroup analyses also identified the type of exercise as a significant (p=0.013) moderator of effect sizes within an exercise time frame of 0.5-10 min with greater effect sizes for exercise capacity (0.4998 (95% CI 0.246 to 0.753)) versus performance (0.1078 (95% CI -0.201 to 0.416)). There was no moderating effect of training status (p=0.559), intermittent or continuous exercise (p=0.436) or total amount of β-alanine ingested (p=0.438). Co-supplementation with sodium bicarbonate resulted in the largest effect size when compared with placebo (0.43 (95% CI 0.22 to 0.64)). β-alanine had a

  17. Glycolysis and the Tricarboxylic Acid Cycle Are Linked by Alanine Aminotransferase during Hypoxia Induced by Waterlogging of Lotus japonicus1[W][OA

    Science.gov (United States)

    Rocha, Marcio; Licausi, Francesco; Araújo, Wagner L.; Nunes-Nesi, Adriano; Sodek, Ladaslav; Fernie, Alisdair R.; van Dongen, Joost T.

    2010-01-01

    The role of nitrogen metabolism in the survival of prolonged periods of waterlogging was investigated in highly flood-tolerant, nodulated Lotus japonicus plants. Alanine production revealed to be a critical hypoxic pathway. Alanine is the only amino acid whose biosynthesis is not inhibited by nitrogen deficiency resulting from RNA interference silencing of nodular leghemoglobin. The metabolic changes that were induced following waterlogging can be best explained by the activation of alanine metabolism in combination with the modular operation of a split tricarboxylic acid pathway. The sum result of this metabolic scenario is the accumulation of alanine and succinate and the production of extra ATP under hypoxia. The importance of alanine metabolism is discussed with respect to its ability to regulate the level of pyruvate, and this and all other changes are discussed in the context of current models concerning the regulation of plant metabolism. PMID:20089769

  18. Activity of the lactate-alanine shuttle is independent of glutamate-glutamine cycle activity in cerebellar neuronal-astrocytic cultures

    DEFF Research Database (Denmark)

    Bak, Lasse K; Sickmann, Helle M; Schousboe, Arne

    2004-01-01

    The glutamate-glutamine cycle describes the neuronal release of glutamate into the synaptic cleft, astrocytic uptake, and conversion into glutamine, followed by release for use as a neuronal glutamate precursor. This only explains the fate of the carbon atoms, however, and not that of the ammonia....... Recently, a role for alanine has been proposed in transfer of ammonia between glutamatergic neurons and astrocytes, denoted the lactate-alanine shuttle (Waagepetersen et al. [ 2000] J. Neurochem. 75:471-479). The role of alanine in this context has been studied further using cerebellar neuronal cultures...... and corresponding neuronal-astrocytic cocultures. A superfusion paradigm was used to induce repetitively vesicular glutamate release by N-methyl-D-aspartate (NMDA) in the neurons, allowing the relative activity dependency of the lactate-alanine shuttle to be assessed. [(15)N]Alanine (0.2 mM), [2-(15)N]/[5-(15)N...

  19. Reconfiguration of N Metabolism upon Hypoxia Stress and Recovery: Roles of Alanine Aminotransferase (AlaAT and Glutamate Dehydrogenase (GDH

    Directory of Open Access Journals (Sweden)

    Houssein Diab

    2016-05-01

    Full Text Available In the context of climatic change, more heavy precipitation and more frequent flooding and waterlogging events threaten the productivity of arable farmland. Furthermore, crops were not selected to cope with flooding- and waterlogging-induced oxygen limitation. In general, low oxygen stress, unlike other abiotic stresses (e.g., cold, high temperature, drought and saline stress, received little interest from the scientific community and less financial support from stakeholders. Accordingly, breeding programs should be developed and agronomical practices should be adapted in order to save plants’ growth and yield—even under conditions of low oxygen availability (e.g., submergence and waterlogging. The prerequisite to the success of such breeding programs and changes in agronomical practices is a good knowledge of how plants adapt to low oxygen stress at the cellular and the whole plant level. In the present paper, we summarized the recent knowledge on metabolic adjustment in general under low oxygen stress and highlighted thereafter the major changes pertaining to the reconfiguration of amino acids syntheses. We propose a model showing (i how pyruvate derived from active glycolysis upon hypoxia is competitively used by the alanine aminotransferase/glutamate synthase cycle, leading to alanine accumulation and NAD+ regeneration. Carbon is then saved in a nitrogen store instead of being lost through ethanol fermentative pathway. (ii During the post-hypoxia recovery period, the alanine aminotransferase/glutamate dehydrogenase cycle mobilizes this carbon from alanine store. Pyruvate produced by the reverse reaction of alanine aminotransferase is funneled to the TCA cycle, while deaminating glutamate dehydrogenase regenerates, reducing equivalent (NADH and 2-oxoglutarate to maintain the cycle function.

  20. Effects of beta-alanine supplementation on brain homocarnosine/carnosine signal and cognitive function: an exploratory study.

    Directory of Open Access Journals (Sweden)

    Marina Yazigi Solis

    Full Text Available Two independent studies were conducted to examine the effects of 28 d of beta-alanine supplementation at 6.4 g d(-1 on brain homocarnosine/carnosine signal in omnivores and vegetarians (Study 1 and on cognitive function before and after exercise in trained cyclists (Study 2.In Study 1, seven healthy vegetarians (3 women and 4 men and seven age- and sex-matched omnivores undertook a brain 1H-MRS exam at baseline and after beta-alanine supplementation. In study 2, nineteen trained male cyclists completed four 20-Km cycling time trials (two pre supplementation and two post supplementation, with a battery of cognitive function tests (Stroop test, Sternberg paradigm, Rapid Visual Information Processing task being performed before and after exercise on each occasion.In Study 1, there were no within-group effects of beta-alanine supplementation on brain homocarnosine/carnosine signal in either vegetarians (p = 0.99 or omnivores (p = 0.27; nor was there any effect when data from both groups were pooled (p = 0.19. Similarly, there was no group by time interaction for brain homocarnosine/carnosine signal (p = 0.27. In study 2, exercise improved cognitive function across all tests (P 0.05 of beta-alanine supplementation on response times or accuracy for the Stroop test, Sternberg paradigm or RVIP task at rest or after exercise.28 d of beta-alanine supplementation at 6.4 g d(-1 appeared not to influence brain homocarnosine/carnosine signal in either omnivores or vegetarians; nor did it influence cognitive function before or after exercise in trained cyclists.

  1. 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.

  2. Reconfiguration of N Metabolism upon Hypoxia Stress and Recovery: Roles of Alanine Aminotransferase (AlaAT) and Glutamate Dehydrogenase (GDH)

    Science.gov (United States)

    Diab, Houssein; Limami, Anis M.

    2016-01-01

    In the context of climatic change, more heavy precipitation and more frequent flooding and waterlogging events threaten the productivity of arable farmland. Furthermore, crops were not selected to cope with flooding- and waterlogging-induced oxygen limitation. In general, low oxygen stress, unlike other abiotic stresses (e.g., cold, high temperature, drought and saline stress), received little interest from the scientific community and less financial support from stakeholders. Accordingly, breeding programs should be developed and agronomical practices should be adapted in order to save plants’ growth and yield—even under conditions of low oxygen availability (e.g., submergence and waterlogging). The prerequisite to the success of such breeding programs and changes in agronomical practices is a good knowledge of how plants adapt to low oxygen stress at the cellular and the whole plant level. In the present paper, we summarized the recent knowledge on metabolic adjustment in general under low oxygen stress and highlighted thereafter the major changes pertaining to the reconfiguration of amino acids syntheses. We propose a model showing (i) how pyruvate derived from active glycolysis upon hypoxia is competitively used by the alanine aminotransferase/glutamate synthase cycle, leading to alanine accumulation and NAD+ regeneration. Carbon is then saved in a nitrogen store instead of being lost through ethanol fermentative pathway. (ii) During the post-hypoxia recovery period, the alanine aminotransferase/glutamate dehydrogenase cycle mobilizes this carbon from alanine store. Pyruvate produced by the reverse reaction of alanine aminotransferase is funneled to the TCA cycle, while deaminating glutamate dehydrogenase regenerates, reducing equivalent (NADH) and 2-oxoglutarate to maintain the cycle function. PMID:27258319

  3. Effect of sodium bicarbonate and Beta-alanine on repeated sprints during intermittent exercise performed in hypoxia.

    Science.gov (United States)

    Saunders, Bryan; Sale, Craig; Harris, Roger C; Sunderland, Caroline

    2014-04-01

    To investigate the separate and combined effects of sodium bicarbonate and beta-alanine supplementation on repeated sprints during simulated match play performed in hypoxia. Study A: 20 recreationally active participants performed two trials following acute supplementation with either sodium bicarbonate (0.3 g·kg-1BM) or placebo (maltodextrin). Study B: 16 recreationally active participants were supplemented with either a placebo or beta-alanine for 5 weeks (6.4 g·day-1 for 4 weeks, 3.2 g·day-1 for 1 week), and performed one trial before supplementation (with maltodextrin) and two following supplementation (with sodium bicarbonate and maltodextrin). Trials consisted of 3 sets of 5 × 6 s repeated sprints performed during a football specific intermittent treadmill protocol performed in hypoxia (15.5% O2). Mean (MPO) and peak (PPO) power output were recorded as the performance measures. Study A: Overall MPO was lower with sodium bicarbonate than placebo (p = .02, 539.4 ± 84.5 vs. 554.0 ± 84.6 W), although there was no effect across sets (all p > .05). Study B: There was no effect of beta-alanine, or cosupplementation with sodium bicarbonate, on either parameter, although there was a trend toward higher MPO with sodium bicarbonate (p = .07). The effect of sodium bicarbonate on repeated sprints was equivocal, although there was no effect of beta-alanine or cosupplementation with sodium bicarbonate. Individual variation may have contributed to differences in results with sodium bicarbonate, although the lack of an effect with beta-alanine suggests this type of exercise may not be influenced by increased buffering capacity.

  4. Stimulation of Na+-alanine cotransport activates a voltage-dependent conductance in single proximal tubule cells isolated from frog kidney

    Science.gov (United States)

    Robson, L; Hunter, M

    1999-01-01

    The swelling induced by Na+-alanine cotransport in proximal tubule cells of the frog kidney is followed by regulatory volume decrease (RVD). This RVD is inhibited by gadolinium (Gd3+), an inhibitor of stretch-activated channels, but is independent of extracellular Ca2+. In this study, the whole cell patch clamp technique was utilized to examine the effect of Na+-alanine cotransport on two previously identified volume- and Gd3+-sensitive conductances. One conductance is voltage dependent and anion selective (GVD) whilst the other is voltage independent and cation selective (GVI). Addition of 5 mM L-alanine to the bathing solution increased the whole cell conductance and gave a positive (depolarizing) shift in the reversal potential (Vrev, equivalent to the membrane potential in current-clamped cells) consistent with activation of Na+-alanine cotransport. Vrev shifted from -36 ± 4·9 to +12·9 ± 4·2 mV (n= 15). In the presence of alanine, the total whole cell conductance had several components including the cotransporter conductance and GVD and GVI. These conductances were separated using Gd3+, which inhibits both GVD and GVI, and the time dependency of GVD. Of these two volume-sensitive conductances, L-alanine elicited a specific increase in GVD, whereas GVI was unaffected. The L-alanine-induced activation of GVD was significantly reduced when cells were incubated in a hypertonic bathing solution. In summary, in single proximal tubule cells isolated from frog kidney, on stimulation of Na+-alanine cotransport GVD is activated, while GVI is unaffected. Taken with other evidence, this suggests that GVD is activated by cell swelling, consequent upon alanine entry, and may play a role as an anion efflux pathway during alanine-induced volume regulation. PMID:10226159

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

    Directory of Open Access Journals (Sweden)

    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

  6. Optical, thermal and magnetic studies of pure and cobalt chloride doped L-alanine cadmium chloride

    Energy Technology Data Exchange (ETDEWEB)

    Benila, B.S., E-mail: benjane.benila@gmail.com [Department of Physics and Research Centre, Scott Christian College (Autonomous), Nagercoil 629 003 (India); Bright, K.C. [Department of Physics, St. John' s College, Anchal, Kollam 691 306 (India); Delphine, S. Mary [Department of Physics, Holy Cross College (Autonomous), Nagercoil 629 004 (India); Shabu, R. [Department of Physics and Research Centre, Scott Christian College (Autonomous), Nagercoil 629 003 (India)

    2017-03-15

    Single crystals of L-alanine cadmium chloride (LACC) and cobalt chloride (Co{sup 2+}) doped LACC have been grown by the slow evaporation solution growth technique. The grown crystals were subjected to various characterizations such as powder XRD, SXRD, FTIR, UV–vis, EDAX, TG/DTA, VSM, Dielectric and Second Harmonic Generation (SHG) measurements. The lattice parameters of the grown crystals were determined by single crystal X-ray analysis. EDAX analysis confirms the presence of Co{sup 2+} ion in the host material. The functional group and optical behavior of the crystals were identified from FTIR and UV-vis spectrum analysis. Electrical parameters such as dielectric constant, dielectric loss have been studied. The thermal stability of the compound was found out using TGA/DTA analysis. Second Harmonic Generation of the samples was confirmed by Kurtz-Perry powder technique. Magnetic properties of the crystals studied by VSM were also reported. The encouraging results show that the cobalt chloride doped LACC crystals have greater potential applications in optical devices. - Graphical abstract: Fig (a) and (b) shows the transparent, stable single crystals of pure and doped crystals were obtained using slow evaporation technique. The sizes of pure and doped crystals are 20×9×2 mm{sup 3} and 18×15×1 mm{sup 3} respectively. Fig (c) is the Hysteresis loop traced at room temperature for the pure and doped crystals explains the soft ferromagnetic nature of the doped crystal. The provision for changing the value of coercivity can be used for security, switching and sensing applications. - Highlights: • Defect free crystals of pure and Co{sup 2+} ion doped L-alanine cadmium chloride were grown. • The optical, dielectric and magnetic properties of pure crystals were enhanced by adding Co{sup 2+} ion. • High optical transmittance was obtained in the entire visible and IR region. • Addition of dopant to the pure crystal altered the coercivity. • Low dielectric

  7. Effect of β-alanine plus sodium bicarbonate on high-intensity cycling capacity.

    Science.gov (United States)

    Sale, Craig; Saunders, Bryan; Hudson, Sean; Wise, John A; Harris, Roger C; Sunderland, Caroline D

    2011-10-01

    We examined the effect of β-alanine supplementation plus sodium bicarbonate on high-intensity cycling capacity. Twenty males (age = 25 ± 5 yr, height = 1.79 ± 0.06 m, body mass = 80.0 ± 10.3 kg) were assigned to either a placebo (P) or a β-alanine (BA; 6.4 g·d(-1) for 4 wk) group based on power max, completing four cycling capacity tests at 110% of power max (CCT110%) to determine time to exhaustion (TTE) and total work done. A CCT(110%) was performed twice (habituation and baseline) before supplementation (with maltodextrin [MD]) and twice after supplementation (with MD and with sodium bicarbonate [SB]), using a crossover design with 2 d of rest between trials, creating four study conditions (PMD, PSB, BAMD, and BASB). Blood pH, Lactate, bicarbonate and base excess were determined at baseline, before exercise, immediately after exercise, and 5 min after exercise. Data were analyzed using repeated-measures ANOVA. TTE was increased in all conditions after supplementation (+1.6% PMD, +6.5% PSB, +12.1% BAMD, and +16.2% BASB). Both BAMD and BASB resulted in significantly improved TTE compared with that before supplementation (P ≤ 0.01). Although further increases in TTE (4.1%) were shown in BASB compared with BAMD, these differences were not significant (P = 0.74). Differences in total work done were similar to those of TTE. Blood bicarbonate concentrations were significantly (P ≤ 0.001) elevated before exercise in PSB and BASB but not in PMD or BAMD. Blood lactate concentrations were significantly elevated after exercise, remaining elevated after 5 min of recovery (P ≤ 0.001) and were highest in PSB and BASB. Results show that BA improved high-intensity cycling capacity. However, despite a 6-s (∼4%) increase in TTE with the addition of SB, this did not reach statistical significance, but magnitude-based inferences suggested a ∼70% probability of a meaningful positive difference.

  8. The effect of isotretinoin on triglycerides and liver aminotransferases Influência da isotretinoína nas transaminases hepáticas e triglicerídeos

    Directory of Open Access Journals (Sweden)

    Andreia Salezze Vieira

    2012-06-01

    Full Text Available BACKGROUND: Isotretinoin has been used to treat the most severe cases of acne; however, it may provoke adverse events in mucocutaneous and hepatic tissues, lead to alterations in lipid levels and cause teratogenicity. OBJECTIVE: The objective of this study was to evaluate the profile of changes in alanine aminotransferase (ALT, aspartate aminotransferase (AST and triglyceride levels in patients who had been treated with oral isotretinoin dispensed by the São Mateus/ES pharmacy for special drugs. METHODS: A retrospective, observational, longitudinal study was conducted by carrying out a secondary analysis of each patient's data. RESULTS: Of the 130 patients who received isotretinoin between January and December 2009, only 70 were actually treated for 3 months or more and handed in the results of their laboratory tests. Of these 70 patients, 39 (55.7% were female. The mean age of the women (23.9 years was higher than the mean age of the men (20.1 years. There was a statistically significant increase in the levels of triglycerides (87.01 ± 48.25 versus 105.32 ± 48.76 mg/dL, AST (20.44 ± 6.26 versus 24.38 ± 11.92 U/L and ALT (18.24 ± 8.31 versus 23.34 ± 20.03 U/L performed prior to and 3 months or more after oral isotretinoin treatment. After treatment with oral isotretinoin, triglyceride levels had increased beyond the normal range in 11% of the patients, while 8.6% had elevated AST levels and 7.3% had increased ALT levels. CONCLUSION: The results in this population show that the use of oral isotretinoin for the treatment of acne may result in altered triglyceride, AST and ALT levels. These findings are in accordance with data published previously in the scientific literature, confirming the need to monitor these patients.FUNDAMENTOS: A isotretinoína tem sido usada no tratamento dos casos mais graves de acne, embora possa induzir reações adversas nos tecidos mucocutâneos e hepáticos, alterações nos níveis lipídicos e

  9. Relationship Between HCV RNA and Serum Alanine Aminotransferase and HCV Genotype Distribution

    Directory of Open Access Journals (Sweden)

    Dilara Yildirim

    2014-03-01

    Full Text Available Aim: Hepatitis C virus (HCV infections is an important health problem all over the world because of inveteracy ratio and causing hepatic failure, cirrhosis and hepatocellular carcinoma. Our aim in this study is to research HCV RNA in the patients detected anti-HCV positivity with enzyme immunoassay (ELISA and to analysis the association of HCV RNA in the serum of patients with alanine aminoransferase (ALT levels and HCV genotypes. Material and Method: In anti-HCV positive patients came from various departments to our laboratory HCV RNA level and genotyping were researched with quantitive real time polimerase chain reaction (PCR Montania 483 (Anatolia Geneworks HCV, Turkey and anti-HCV tests were studied with ELISA (Advia Centaur XP, Germany, ALT results were analysed retrospectively (Olympus AU2700. Results: In 123 of Anti HCV positive 338 serum (%37 were detected HCV RNA positivity. In the patients who have ALT level more than 40 (%77.6, HCV RNA is positive. Relationship between HCVRNA positive patients and the patients who have ALT level more than 40 is meaningful. (p

  10. Consequences of proline-to-alanine substitutions for the stability and refolding of onconase.

    Science.gov (United States)

    Hacke, Mandy; Gruber, Tobias; Schulenburg, Cindy; Balbach, Jochen; Arnold, Ulrich

    2013-09-01

    Peptidyl-prolyl isomerization reactions can make for rate-limiting steps in protein folding due to their high activation energy. Onconase, an unusually stable ribonuclease A homologue from the Northern leopard frog, contains four trans proline residues in its native state. During the refolding from its guanidine hydrochloride unfolded state, which includes the formation of a folding intermediate, the slowest of the three phases has earlier been attributed to a cis-to-trans peptidyl-prolyl isomerization reaction. We thus substituted all four proline residues individually by alanine and investigated the effect of the amino acid substitutions on the folding and stability of the onconase variants. All onconase variants proved to adopt a tertiary structure comparable with that of the wild-type protein. Although the slow phase was not eliminated for any of the variants, the P43A substitution resulted in an increase in the rate constant of the fast folding phase, i.e. a faster formation of the folding intermediate. This variant also exhibits a significant increase in thermodynamic stability. As residue 43 belongs to those residues that are protected from hydrogen exchange with the solvent in the folding intermediate, the increase in the rate constant and stability of the P43A variant emphasizes the importance of the intermediate for the folding of onconase. © 2013 FEBS.

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

    Directory of Open Access Journals (Sweden)

    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.

  12. Effect of radiation quality on radical formation in ion-irradiated solid alanine

    Energy Technology Data Exchange (ETDEWEB)

    Koizumi, Hitoshi; Ichikawa, Tsuneki; Yoshida, Hiroshi [Hokkaido Univ., Sapporo (Japan); Namba, Hideki; Taguchi, Mitsumasa; Kojima, Takuji

    1997-03-01

    Radical formation in solid alanine irradiated with H{sup +} and He{sup +} ions of 0.5-3.0 MeV and with heavy ions of hundreds of MeV was examined by the ESR method. Radical yield is constant below a critical fluence, and the yield decreases above the fluence. The critical fluence for the H{sup +} and He{sup +} ions is about 10{sup 12} ions cm{sup -2}, while the critical fluence for the heavy ions is 10{sup 10}-10{sup 11} ions cm{sup -2}. G-value of the radical formation (radicals per 100 eV absorbed dose) is obtained from the constant yield at the low fluences. The G-value depends on the radiation quality. This dependence is ascribed to the difference of local dose in the ion tracks. The fluence-yield curves were simulated with a model assuming cylindrical shape of ion tracks and dose-yield relationship for {gamma}-irradiation. This model well explains the fluence-yield curves for the ion irradiations. (author)

  13. A Micro-Platinum Wire Biosensor for Fast and Selective Detection of Alanine Aminotransferase.

    Science.gov (United States)

    Thuy, Tran Nguyen Thanh; Tseng, Tina T-C

    2016-05-26

    In this study, a miniaturized biosensor based on permselective polymer layers (overoxidized polypyrrole (Ppy) and Nafion(®)) modified and enzyme (glutamate oxidase (GlutOx)) immobilized micro-platinum wire electrode for the detection of alanine aminotransferase (ALT) was fabricated. The proposed ALT biosensor was measured electrochemically by constant potential amperometry at +0.7 V vs. Ag/AgCl. The ALT biosensor provides fast response time (~5 s) and superior selectivity towards ALT against both negatively and positively charged species (e.g., ascorbic acid (AA) and dopamine (DA), respectively). The detection range of the ALT biosensor is found to be 10-900 U/L which covers the range of normal ALT levels presented in the serum and the detection limit and sensitivity are found to be 8.48 U/L and 0.059 nA/(U/L·mm²) (N = 10), respectively. We also found that one-day storage of the ALT biosensor at -20 °C right after the sensor being fabricated can enhance the sensor sensitivity (1.74 times higher than that of the sensor stored at 4 °C). The ALT biosensor is stable after eight weeks of storage at -20 °C. The sensor was tested in spiked ALT samples (ALT activities: 20, 200, 400, and 900 U/L) and reasonable recoveries (70%~107%) were obtained.

  14. Investigation of the interaction of β-methylamino-L-alanine with eukaryotic and prokaryotic proteins.

    Science.gov (United States)

    Main, Brendan J; Italiano, Carly J; Rodgers, Kenneth J

    2017-12-12

    There is a strong body of evidence linking the non-protein amino acid (NPAA) β-methylamino-L-alanine (BMAA) to the development of a number of neurodegenerative diseases. BMAA has been found globally, is produced by a number of organisms including cyanobacteria, diatoms, and dinoflagellates; and has been shown to biomagnify through trophic levels. The role of BMAA in neurodegenerative disease is highlighted by its presence in the brains of a number of neurodegenerative disease patients, where it was found in a protein-bound form. We have previously shown that BMAA is bound to cell proteins, and results in the upregulation of the unfolded protein response, an endoplasmic reticulum stress response activated by the presence of misfolded proteins within the cell. Structurally aberrant proteins are features of a number of neurodegenerative diseases, and further investigation of how BMAA interacts with proteins is crucial to our understanding of its toxicity. Here we use radiolabelled BMAA to investigate the interaction and binding of BMAA to eukaryotic and prokaryotic proteins. We found differences in the presence and distribution of protein-bound BMAA between E. coli and neuroblastoma cells, with an increase in binding over time only seen in the eukaryotic cells. We also found that BMAA was unable to bind to pure proteins, or cell lysate in native or denaturing conditions, indicating that biological processing is required for BMAA to bind to proteins.

  15. Analysis of the enzymatic properties of a broad family of alanine aminotransferases.

    Directory of Open Access Journals (Sweden)

    Chandra H McAllister

    Full Text Available Alanine aminotransferase (AlaAT has been studied in a variety of organisms due to the involvement of this enzyme in mammalian processes such as non-alcoholic hepatocellular damage, and in plant processes such as C4 photosynthesis, post-hypoxic stress response and nitrogen use efficiency. To date, very few studies have made direct comparisons of AlaAT enzymes and fewer still have made direct comparisons of this enzyme across a broad spectrum of organisms. In this study we present a direct kinetic comparison of glutamate:pyruvate aminotransferase (GPAT activity for seven AlaATs and two glutamate:glyoxylate aminotransferases (GGAT, measuring the K(M values for the enzymes analyzed. We also demonstrate that recombinant expression of AlaAT enzymes in Eschericia coli results in differences in bacterial growth inhibition, supporting previous reports of AlaAT possessing bactericidal properties, attributed to lipopolysaccharide endotoxin recognition and binding. A probable lipopolysaccharide binding region within the AlaAT enzymes, homologous to a region of a lipopolysaccharide binding protein (LBP in humans, was also identified in this study. The AlaAT enzyme differences identified here indicate that AlaAT homologues have differentiated significantly and the roles these homologues play in vivo may also have diverged significantly. Specifically, the differing kinetics of AlaAT enzymes and how this may alter the nitrogen use efficiency in plants is discussed.

  16. Prevalence and predictors of alanine aminotransferase elevation among normal weight, overweight and obese youth in Mexico.

    Science.gov (United States)

    Purcell, Maura; Flores, Yvonne N; Zhang, Zuo-Feng; Denova-Gutiérrez, Edgar; Salmeron, Jorge

    2013-09-01

    This study aimed to determine the prevalence of and risk factors associated with elevated alanine aminotransferase (ALT) levels among a sample of normal weight, overweight and obese youth from two urban populations in Central Mexico. Baseline data from 1262 youth aged 8-19 years who participated in the Mexican Health Worker Cohort Study from March 2004 to April 2006 were reviewed, including 680 girls and 582 boys, with a total of 83 participants with elevated ALT level (>40 U/L). Information was obtained from self-administered questionnaires, anthropometric results and clinical measurements. Associations of interest were examined using multivariate logistic regression models. A total of 3.8% of girls and 9.8% of boys had elevated ALT levels. Elevated ALT was observed in 28.9% of the obese and 14.2% of the overweight participants. Metabolic syndrome (MS) occurred in 6.1% of the study population and those with MS had a high percentage of elevated ALT (14.5% of girls and 40.0% of boys, respectively). Abdominal obesity and insulin resistance were also associated with a greater risk of elevated ALT. Obesity and certain metabolic risk factors are important predictors for elevated ALT. Screening for ALT levels in obese youth could help to identify those at risk and reduce the possibility of future liver diseases. © 2013 Wiley Publishing Asia Pty Ltd and Chinese Medical Association Shanghai Branch, Chinese Society of Gastroenterology, Renji Hospital Affiliated to Shanghai Jiaotong University School of Medicine.

  17. Optimization of partial multicanonical molecular dynamics simulations applied to an alanine dipeptide in explicit water solvent.

    Science.gov (United States)

    Okumura, Hisashi

    2011-01-07

    The partial multicanonical algorithm for molecular dynamics and Monte Carlo simulations samples a wide range of an important part of the potential energy. Although it is a strong technique for structure prediction of biomolecules, the choice of the partial potential energy has not been optimized. In order to find the best choice, partial multicanonical molecular dynamics simulations of an alanine dipeptide in explicit water solvent were performed with 15 trial choices for the partial potential energy. The best choice was found to be the sum of the electrostatic, Lennard-Jones, and torsion-angle potential energies between solute atoms. In this case, the partial multicanonical simulation sampled all of the local-minimum free-energy states of the P(II), C(5), α(R), α(P), α(L), and C states and visited these states most frequently. Furthermore, backbone dihedral angles ϕ and ψ rotated very well. It is also found that the most important term among these three terms is the electrostatic potential energy and that the Lennard-Jones term also helps the simulation to overcome the steric restrictions. On the other hand, multicanonical simulation sampled all of the six states, but visited these states fewer times. Conventional canonical simulation sampled only four of the six states: The P(II), C(5), α(R), and α(P) states.

  18. Development of an alanine dosimeter for gamma dosimetry in mixed environments

    International Nuclear Information System (INIS)

    Vehar, D.W.; Griffin, P.J.

    1992-01-01

    L-αa-Alanine, a nontoxic polycrystalline amino acid, has been investigated for use in high-precision, high-level absorbed-dose measurements in mixed neutron/photon environments such as research and test reactors. The technique is based on the use of electron paramagnetic resonance (EPR) spectroscopy to determine the extent of free radical production in a sample exposed to ionizing radiation, and has been successfully used for photon absorbed-dose measurements at levels exceeding 10 5 Gy with high measurement precision. Application of the technique to mixed environments requires knowledge of the energy-dependent response of the dosimeter for both photons and neutrons. Determination of the dosimeter response to photons is accomplished by irradiations in 60 Co and bremsstrahlung sources and by calculations of energy-dependent photon kerma. Neutron response is determined by calculations in conjunction with CaF 2 :Mn thermoluminescence dosimeters and by calculations of energy-dependent neutron kerma. Several neutron environments are used, including the ACRR and SPR-III reactors

  19. Development of an alanine dosimeter for gamma dosimetry in mixed environments -- Summary of research

    International Nuclear Information System (INIS)

    Vehar, D.W.; Griffin, P.J.

    1994-02-01

    L-α-alanine, a nontoxic polycrystalline amino acid, has been investigated for use in high-precision, high-level absorbed-dose measurements in mixed neutron/photon environments such as research and test reactors. The technique is based on the use of electron paramagnetic resonance spectroscopy to determine the extent of free radical production in a sample exposed to ionizing radiation, and has been successfully used for photon absorbed-dose measurements at levels exceeding 10 5 Gy with high measurement precision. Application of the technique to mixed environments requires knowledge of the energy-dependent response of the dosimeter for both photons and neutrons. Determination of the dosimeter response to photons is accomplished by irradiations in 60 Co and bremsstrahlung sources and by calculations of energy-dependent photon kerma. Neutron response is determined by irradiations in conjunction with CaF 2 :Mn thermoluminescence dosimeters and by calculations of energy-dependent neutron kerma. Several neutron environments are used, including those provided by the Annular Core Research Reactor and Sandia Pulsed Reactor

  20. Small-Field Dosimetry in A 6 MV Photon Beam Using Alanine and Liquid Ionisation Chamber

    DEFF Research Database (Denmark)

    Zimmermann, S.; Riis, H. L.; Hjelm-Hansen, M.

    2012-01-01

    , 2 times 4 sweeps with a 90° turn in between, total measurement time was 3 minutes per dosimeter. The alanine dosimetry system was calibrated using a Co60 gamma cell ensuring traceability to a national standard. Results: The results are summarized in Table 1. At depths below 2 mm, we noted....... In addition, the measurements were carried out at seven symmetric field sizes with the length of the field edges at the water surface: 0.8, 1.0, 1.4, 2.0, 3.0, 4.0 and 10.0 cm. As a minimum three dosimeters were irradiated to 2500 MU in each configuration. The dosimeters were placed in the dose maximum...... of each field and depth. This dose maximum was measured for each field using a Scanditronix Wellhöfer photon field diode. The same measurements were carried out using a liquid ionchamber, PTW microLion, irradiated by 500 MU. The output of the accelerator was controlled by a PTW semiflex ion chamber...