DOCSLIB.ORG

2206.Full.Pdf

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text

Load more

2206 Diabetes Volume 69, October 2020 Association of the 1q25 Diabetes-Specific Coronary Heart Disease Locus With Alterations of the g-Glutamyl Cycle and Increased Methylglyoxal Levels in Endothelial Cells Caterina Pipino,1,2,3 Hetal Shah,1,2 Sabrina Prudente,4 Natalia Di Pietro,3 Lixia Zeng,5 Kyoungmin Park,1,2 Vincenzo Trischitta,6,7 Subramanian Pennathur,5 Assunta Pandolfi,3 and Alessandro Doria1,2 Diabetes 2020;69:2206–2216 | https://doi.org/10.2337/db20-0475 A chromosome 1q25 variant (rs10911021) has been suggesting that such treatment may prevent CHD in 1q25 associated with coronary heart disease (CHD) in type risk allele carriers. 2 diabetes. In human umbilical vein endothelial cells (HUVECs), the risk allele “C” is associated with lower expression of the adjacent gene GLUL encoding gluta- Despite improvements in glycemic control and other pre- mine synthase, converting glutamic acid to glutamine. To ventive therapies, individuals with type 2 diabetes con- further investigate the mechanisms through which this tinue to experience an increased burden of cardiovascular locus affects CHD risk, we measured 35 intracellular complications such as coronary heart disease (CHD) (1). In metabolites involved in glutamic acid metabolism and order to change the status quo, new interventions are the g-glutamyl cycle in 62 HUVEC strains carrying dif- needed that specifically target the mechanisms linking the ferent rs10911021 genotypes. Eight metabolites were diabetic milieu to vascular damage. Our strategy to gain positively associated with the risk allele (17–58% in- insights into these mechanisms and identify novel targets crease/allele copy, P 5 0.046–0.002), including five for preventive therapies has been to study the genetic g-glutamyl amino acids, b-citryl-glutamate, N-acetyl- factors that modulate cardiovascular risk among people aspartyl-glutamate, and ophthalmate—a marker of with type 2 diabetes (2). g-glutamyl cycle malfunction. Consistent with these find- Following this approach, we have discovered a locus ings, the risk allele was also associated with decreased on chromosome 1q25 that is associated with CHD, with glutathione-to-glutamate ratio (29%, P 5 0.012), de- genome-wide significance, in multiple sets of patients creased S-lactoylglutathione (241%, P 5 0.019), and with type 2 diabetes (3). Subsequent studies have con- reduced detoxification of the atherogenic compound firmed this finding in other populations with type 2 di- GENETICS/GENOMES/PROTEOMICS/METABOLOMICS methylglyoxal (154%, P 5 0.008). GLUL downregulation abetes (4,5). This CHD locus is placed in the region of the by shRNA caused a 40% increase in the methylglyoxal GLUL gene, which codes for glutamate-ammonia ligase level, which was completely prevented by glutamine (also known as glutamine synthase) catalyzing the con- supplementation. In summary, we have identified in- version of glutamic acid to glutamine (6). In the original tracellular metabolic traits associated with the 1q25 report, the risk allele of the lead variant (rs10911021) risk allele in HUVECs, including impairments of the was associated with decreased GLUL expression in endo- g-glutamyl cycle and methylglyoxal detoxification. Glu- thelial cells and with a lower pyroglutamic-to-glutamic tamine supplementation abolishes the latter abnormality, acid ratio in plasma, suggesting an impairment of glutamic 1Research Division, Joslin Diabetes Center, Boston, MA Corresponding author: Alessandro Doria, [email protected] 2 Department of Medicine, Harvard Medical School, Boston, MA Received 5 May 2020 and accepted 3 July 2020 3Department of Medical, Oral and Biotechnological Sciences, Center for Advanced This article contains supplementary material online at https://doi.org/10.2337/ Studies and Technology - CAST (ex CeSI-MeT), University G. d’Annunzio of Chieti- figshare.12616442. Pescara, Chieti, Italy 4Research Unit of Metabolic and Cardiovascular Diseases, Fondazione IRCCS Casa © 2020 by the American Diabetes Association. Readers may use this article as Sollievo della Sofferenza, San Giovanni Rotondo, Italy long as the work is properly cited, the use is educational and not for profit, and the 5Division of Nephrology, Department of Medicine, University of Michigan, Ann Arbor, MI work is not altered. More information is available at https://www.diabetesjournals 6Research Unit of Diabetes and Endocrine Diseases, Fondazione IRCCS Casa Sollievo .org/content/license. della Sofferenza, San Giovanni Rotondo, Italy 7Department of Experimental Medicine, Sapienza University, Rome, Italy diabetes.diabetesjournals.org Pipino and Associates 2207 acid metabolism and the g-glutamyl cycle, of which Genotyping pyroglutamic acid is an intermediate, as a possible mech- DNA was extracted from HUVECs by standard methods. anism underlying the association with CHD (3). The Genotyping of the rs10911021 variant was performed g-glutamyl cycle is responsible for the generation of by means of a custom TaqMan assay (Life Technologies, glutathione—a natural antioxidant playing a critical Foster City, CA) on a 7900HT platform (Applied Biosys- role in the protection from free radicals and other tems, Foster City, CA). Genotyping quality was tested by reactive compounds (7)—whose deficit may predispose including six blinded duplicate samples in each 96-well to increased oxidative stress and accelerated athero- assay. The average agreement rate of the duplicate samples . sclerosis (7,8). was 99%. The goal of the current study was to seek further Metabolomic Profiling support for this hypothesis by analyzing the association at the cellular level between the 1q25 lead variant Cell Culture – fl (rs10911021) and metabolites involved in glutamic acid HUVECs at 60 70% con uence were incubated with media metabolism and the g-glutamyl cycle in a unique collection containing 10% FBS and 5.5 mmol/L D-glucose (basal of human umbilical vein endothelial cells (HUVECs) iso- condition, low glucose) or 25 mmol/L D-glucose (high lated from a large group of newborns and therefore glucose). After 48 h of treatment, cells were trypsinized, allowing the study of the impact of natural genetic transferred to a 50-mL polypropylene Falcon tube, and centrifuged at 1,200 rpm for 10 min. Supernatants were variation on cellular functions. Our results support the discarded and cell pellets gently resuspended in PBS and hypothesis of an association between the 1q25 locus and transferred to prelabeled 2.0-mL polypropylene tubes. Cell alternations in the g-glutamyl cycle and point to a reduced suspensions were centrifuged (1,200 rpm for 10 min) and glutathione-mediated detoxification of methylglyoxal—a all supernatants carefully removed. Cell pellets were im- precursor of advanced glycation end products (AGEs)—as mediately stored at 280°C until all biological replicates a possible mechanism linking these metabolic changes were collected and shipped to Metabolon (Durham, NC) to the increased CHD risk experienced by 1q25 risk allele for metabolomic profiling. carriers with type 2 diabetes. Metabolite Measurements RESEARCH DESIGN AND METHODS The current study considered 35 metabolites involved in HUVEC Preparation and Culture glutamic acid metabolism, the g-glutamyl cycle, and glu- The study was performed on HUVECs isolated from the tathione metabolism that were extracted from a larger umbilical cords of 62 newborns delivered by randomly Metabolon panel of 597 metabolites according to the fi selected, healthy Caucasian mothers between the 36th and prespeci ed hypothesis of an impact of the 1q25 locus on these metabolic pathways. These 35 metabolites included the 40th gestational week at the Hospital of Chieti and all those whose “Sub-pathway” field in the Metabolon Pescara (Italy). HUVECs were isolated from the umbilical report was labeled as “glutathione metabolism” (n 5 9), cords by extraction with 1 mg/mL collagenase 1A at 37°C, “glutamate metabolism” (n 5 13), or “g-glutamyl amino as previously described (9). All procedures were in agree- acid” (n 5 10), plus cysteine and glycine (the two amino ment with the ethical standards of the local Institutional acids that are used together with glutamic acid to synthe- Committee on Human Experimentation (reference size glutathione) and a-ketoglutarate, a Krebs cycle in- number: 1879/09COET) (University G. d’Annunzio of termediate that is in direct equilibrium with glutamic acid Chieti-Pescara, Chieti, Italy) and with the Declaration through amination/deamination by glutamate dehydroge- of Helsinki ethical principles. After approval of the nase. Samples were prepared by Metabolon using the protocol by the Institutional Review Board, signed in- automated MicroLab STAR system from Hamilton formed consent was obtained from each participating Company. Several recovery standards were added before subject. the first step in the extraction process for quality control For this study, HUVECs were grown on 1.5% gelatin- purposes. coated tissue culture plates (Sigma-Aldrich) in complete To remove proteins, dissociate small molecules bound low-glucose (1 g/L) culture medium composed of DMEM to proteins or trapped in the precipitated protein matrix, (provided by Joslin Media Core) supplemented with 20% and to recover chemically diverse metabolites, proteins FBS (Gibco by Life Technologies), 200 units/mL penicillin, were precipitated with methanol under vigorous shaking 200 mg/mL streptomycin (Sigma-Aldrich), 10 mg/mL hep- for 2 min (GenoGrinder 2000; Glen Mills), followed by arin (Stem Cell Technologies), and 50 mg/mL endothelial
Recommended publications
  • Mechanistic Insights on the Reduction of Glutathione Disulfide by Protein Disulfide Isomerase

    Mechanistic Insights on the Reduction of Glutathione Disulfide by Protein Disulfide Isomerase

    Mechanistic insights on the reduction of glutathione disulfide by protein disulfide isomerase Rui P. P. Nevesa, Pedro Alexandrino Fernandesa, and Maria João Ramosa,1 aUnidade de Ciências Biomoleculares Aplicadas, Rede de Química e Tecnologia, Departamento de Química e Bioquímica, Faculdade de Ciências, Universidade do Porto, 4169-007 Porto, Portugal Edited by Donald G. Truhlar, University of Minnesota, Minneapolis, MN, and approved May 9, 2017 (received for review November 22, 2016) We explore the enzymatic mechanism of the reduction of glutathione of enzymes, which are responsible for the reduction and isomer- disulfide (GSSG) by the reduced a domain of human protein disulfide ization of disulfide bonds, through thiol-disulfide exchange. isomerase (hPDI) with atomistic resolution. We use classical molecular dynamics and hybrid quantum mechanics/molecular mechanics cal- Structure and Function of PDI culations at the mPW1N/6–311+G(2d,2p):FF99SB//mPW1N/6–31G(d): Human protein disulfide isomerase (hPDI) is a U-shaped enzyme FF99SB level. The reaction proceeds in two stages: (i) a thiol-disulfide with 508 residues. Its tertiary structure is composed of four exchange through nucleophilic attack of the Cys53-thiolate to the thioredoxin-like domains (a, b, b′,anda′) and a fifth tail-shaped c GSSG-disulfide followed by the deprotonation of Cys56-thiol by domain (Fig. 1) (14, 15). The maximum activity of hPDI is observed Glu47-carboxylate and (ii) a second thiol-disulfide exchange between when all domains of PDI contribute synergistically to its function (16). the Cys56-thiolate and the mixed disulfide intermediate formed in Similar to thioredoxin, the a and a′ domains have a catalytic the first step.
  • Degradation of Glutathione in Plant Cells

    Degradation of Glutathione in Plant Cells

    Degradation of Glutathione in Plant Cells: Evidence against the Participation of a y-Glutamyltranspeptidase Reinhard Steinkamp and Heinz Rennenberg Botanisches Institut der Universität zu Köln, Gyrhofstr. 15, D-5000 Köln 41, Bundesrepublik Deutschland Z. Naturforsch. 40c, 29 — 33 (1985); received August 31/October 4, 1984 Tobacco, Glutathione Catabolism, y-Glutamylcysteine, y-Glutamyltranspeptidase, y-Glutamyl- cyclotransferase When y-glutamyltranspeptidase activity in tobacco cells was measured using the artificial substrate y-glutamyl-/?-nitroanilide, liberation of p-nitroaniline was not reduced, but stimulated by addition of glutathione. Therefore, glutathione was not acting as a donator, but as an acceptor of y-glutamyl moieties in the assay mixture, suggesting that y-glutamyltranspeptidase is not participating in degradation of glutathione. Feeding experiments with [^S-cysJglutathione sup­ ported this conclusion. When tobacco cells were supplied with this peptide as sole sulfur source, glutathione and y-glutamylcysteine were the only labelled compounds found inside the cells. The low rate of uptake of glutathione apparently prevented the accumulation of measurable amounts of radioactivity in the cysteine pool. A y-glutamylcyclotransferase, responsible for the conversion of y-glutamylcysteine to 5-oxo-proline and cysteine was found in ammonium sulfate precipitates of tobacco cell homogenates. The enzyme showed high activities with y-glutamylmethionine and y-glutamylcysteine, but not with other y-glutamyldipeptides or glutathione. From these and previously published experiments [(Rennenberg et al., Z. Naturforsch. 3 5 c, 70 8 -7 1 1 (1980)], it is concluded that glutathione is degraded in tobacco cells via the following pathway: y-glu-cys- gly —> y-glu-cys ->• 5-oxo-proline -* glu. Introduction the cysteine conjugate by the action of a y-gluta­ myltranspeptidase (Fig.
  • Sulfhydryl Reduction of Methylene Blue with Reference to Alterations in Malignant Neoplastic Disease

    Sulfhydryl Reduction of Methylene Blue with Reference to Alterations in Malignant Neoplastic Disease

    Sulfhydryl Reduction of Methylene Blue With Reference to Alterations in Malignant Neoplastic Disease Maurice M. Black, M. D. (From the Department of Biochemistry, New York Medical College, New York 29, N. t;., and the Brooklyn Cancer Institute, Brooklyn 9, N. Y.) (Received for publication May 8, 1947) A significant decrease in methylene blue re- reactivity is less than half that of the cysteine. It is ducing power of plasma from patients with malig- noteworthy also that the resultant leuco mixture nant neoplastic disease was previously reported did not revert back to colored methylene blue on (1). At that time it was suggested that change in a cooling, as was the case with methylene blue re- reducing group of the albumin molecule was a duction by plasma. likely source of this alteration. Similar conclusions Similar relationships were investigated between were reported also by Savignac and associates (7) cysteine and different concentrations of methylene as the result of analogous studies. blue. As seen in Fig. 2, similar curves are obtained, In an attempt to evaluate the effect of the sulf- but the position of the curve on the graph varies hydryl group on the reduction of methylene blue, a with the concentration of the methylene blue used. study was undertaken with various compounds of It should be noted that there is no appreciable known -SH and S-S structures. In addition, an difference in the reducing time of methylene blue attempt was made to establish a standard method on varying the concentrations between 0.10 per of calibration of various lots of methylene blue, so cent and 0.2 per cent, although 0.08 per cent shows that more uniform results would be possible in the a decided difference.
  • A Review of Dietary (Phyto)Nutrients for Glutathione Support

    A Review of Dietary (Phyto)Nutrients for Glutathione Support

    nutrients Review A Review of Dietary (Phyto)Nutrients for Glutathione Support Deanna M. Minich 1,* and Benjamin I. Brown 2 1 Human Nutrition and Functional Medicine Graduate Program, University of Western States, 2900 NE 132nd Ave, Portland, OR 97230, USA 2 BCNH College of Nutrition and Health, 116–118 Finchley Road, London NW3 5HT, UK * Correspondence: [email protected] Received: 8 July 2019; Accepted: 23 August 2019; Published: 3 September 2019 Abstract: Glutathione is a tripeptide that plays a pivotal role in critical physiological processes resulting in effects relevant to diverse disease pathophysiology such as maintenance of redox balance, reduction of oxidative stress, enhancement of metabolic detoxification, and regulation of immune system function. The diverse roles of glutathione in physiology are relevant to a considerable body of evidence suggesting that glutathione status may be an important biomarker and treatment target in various chronic, age-related diseases. Yet, proper personalized balance in the individual is key as well as a better understanding of antioxidants and redox balance. Optimizing glutathione levels has been proposed as a strategy for health promotion and disease prevention, although clear, causal relationships between glutathione status and disease risk or treatment remain to be clarified. Nonetheless, human clinical research suggests that nutritional interventions, including amino acids, vitamins, minerals, phytochemicals, and foods can have important effects on circulating glutathione which may translate to clinical benefit. Importantly, genetic variation is a modifier of glutathione status and influences response to nutritional factors that impact glutathione levels. This narrative review explores clinical evidence for nutritional strategies that could be used to improve glutathione status.
  • Muscle Wasting and Aging: Experimental Models, Fatty Infiltrations, and Prevention Thomas Brioche, Allan Pagano, Guillaume Py, Angèle Chopard

    Muscle Wasting and Aging: Experimental Models, Fatty Infiltrations, and Prevention Thomas Brioche, Allan Pagano, Guillaume Py, Angèle Chopard

    Muscle wasting and aging: Experimental models, fatty infiltrations, and prevention Thomas Brioche, Allan Pagano, Guillaume Py, Angèle Chopard To cite this version: Thomas Brioche, Allan Pagano, Guillaume Py, Angèle Chopard. Muscle wasting and aging: Experi- mental models, fatty infiltrations, and prevention. Molecular Aspects of Medicine, Elsevier, 2016,32 p. 10.1016/j.mam.2016.04.006. hal-01837630 HAL Id: hal-01837630 https://hal.archives-ouvertes.fr/hal-01837630 Submitted on 28 May 2020 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Distributed under a Creative Commons Attribution - ShareAlike| 4.0 International License Accepted Manuscript Title: Muscle wasting and aging: experimental models, fatty infiltrations, and prevention Author: Thomas Brioche, Allan F. Pagano, Guillaume Py, Angèle Chopard PII: S0098-2997(15)30021-2 DOI: http://dx.doi.org/doi: 10.1016/j.mam.2016.04.006 Reference: JMAM 642 To appear in: Molecular Aspects of Medicine Received date: 19-12-2015 Revised date: 13-4-2016 Accepted date: 13-4-2016 Please cite this article as: Thomas Brioche, Allan F. Pagano, Guillaume Py, Angèle Chopard, Muscle wasting and aging: experimental models, fatty infiltrations, and prevention, Molecular Aspects of Medicine (2016), http://dx.doi.org/doi: 10.1016/j.mam.2016.04.006.
  • L -Glutamic Acid (G1251)

    L -Glutamic Acid (G1251)

    L-Glutamic acid Product Number G 1251 Store at Room Temperature Product Description Precautions and Disclaimer Molecular Formula: C5H9NO4 For Laboratory Use Only. Not for drug, household or Molecular Weight: 147.1 other uses. CAS Number: 56-86-0 pI: 3.081 Preparation Instructions 1 pKa: 2.10 (α-COOH), 9.47 (α-NH2), 4.07 (ϕ-COOH) This product is soluble in 1 M HCl (100 mg/ml), with 2 Specific Rotation: D +31.4 ° (6 N HCl, 22.4 °C) heat as needed, yielding a clear, colorless solution. Synonyms: (S)-2-aminoglutaric acid, (S)-2- The solubility in water at 25 °C has been reported to aminopentanedioic acid, 1-aminopropane-1,3- be 8.6 mg/ml.2 dicarboxylic acid, Glu2 Storage/Stability L-Glutamic acid is one of the two amino acids that Aqueous glutamic acid solutions will form contains a carboxylic acid group in its side chains. pyrrolidonecarboxylic acid slowly at room temperature Glutamic acid is commonly referred to as "glutamate", and more rapidly at 100 °C.9 because its carboxylic acid side chain will be deprotonated and thus negatively charged in its References anionic form at physiological pH. In amino acid 1. Molecular Biology LabFax, Brown, T. A., ed., BIOS metabolism, glutamate is formed from the transfer of Scientific Publishers Ltd. (Oxford, UK: 1991), p. amino groups from amino acids to α-ketoglutarate. It 29. thus acts as an intermediary between ammonia and 2. The Merck Index, 12th ed., Entry# 4477. the amino acids in vivo. Glutamate is converted to 3. Biochemistry, 3rd ed., Stryer, L., W.
  • Solutions to 7.012 Problem Set 1

    Solutions to 7.012 Problem Set 1

    MIT Biology Department 7.012: Introductory Biology - Fall 2004 Instructors: Professor Eric Lander, Professor Robert A. Weinberg, Dr. Claudette Gardel Solutions to 7.012 Problem Set 1 Question 1 Bob, a student taking 7.012, looks at a long-standing puddle outside his dorm window. Curious as to what was growing in the cloudy water, he takes a sample to his TA, Brad Student. He wanted to know whether the organisms in the sample were prokaryotic or eukaryotic. a) Give an example of a prokaryotic and a eukaryotic organism. Prokaryotic: Eukaryotic: All bacteria Yeast, fungi, any animial or plant b) Using a light microscope, how could he tell the difference between a prokaryotic organism and a eukaryotic one? The resolution of the light microscope would allow you to see if the cell had a true nucleus or organelles. A cell with a true nucleus and organelles would be eukaryotic. You could also determine size, but that may not be sufficient to establish whether a cell is prokaryotic or eukaryotic. c) What additional differences exist between prokaryotic and eukaryotic organisms? Any answer from above also fine here. In addition, prokaryotic and eukaryotic organisms differ at the DNA level. Eukaryotes have more complex genomes than prokaryotes do. Question 2 A new startup company hires you to help with their product development. Your task is to find a protein that interacts with a polysaccharide. a) You find a large protein that has a single binding site for the polysaccharide cellulose. Which amino acids might you expect to find in the binding pocket of the protein? What is the strongest type of interaction possible between these amino acids and the cellulose? Cellulose is a polymer of glucose and as such has many free hydroxyl groups.
  • Figure S1. Heat Map of R (Pearson's Correlation Coefficient)

    Figure S1. Heat Map of R (Pearson's Correlation Coefficient)

    Figure S1. Heat map of r (Pearson’s correlation coefficient) value among different samples including replicates. The color represented the r value. Figure S2. Distributions of accumulation profiles of lipids, nucleotides, and vitamins detected by widely-targeted UPLC-MC during four fruit developmental stages. The colors indicate the proportional content of each identified metabolites as determined by the average peak response area with R scale normalization. PS1, 2, 3, and 4 represents fruit samples collected at 27, 84, 125, 165 Days After Anthesis (DAA), respectively. Three independent replicates were performed for each stages. Figure S3. Differential metabolites of PS2 vs PS1 group in flavonoid biosynthesis pathway. Figure S4. Differential metabolites of PS2 vs PS1 group in phenylpropanoid biosynthesis pathway. Figure S5. Differential metabolites of PS3 vs PS2 group in flavonoid biosynthesis pathway. Figure S6. Differential metabolites of PS3 vs PS2 group in phenylpropanoid biosynthesis pathway. Figure S7. Differential metabolites of PS4 vs PS3 group in biosynthesis of phenylpropanoids pathway. Figure S8. Differential metabolites of PS2 vs PS1 group in flavonoid biosynthesis pathway and phenylpropanoid biosynthesis pathway combined with RNA-seq results. Table S1. A total of 462 detected metabolites in this study and their peak response areas along the developmental stages of apple fruit. mix0 mix0 mix0 Index Compounds Class PS1a PS1b PS1c PS2a PS2b PS2c PS3a PS3b PS3c PS4a PS4b PS4c ID 1 2 3 Alcohols and 5.25E 7.57E 5.27E 4.24E 5.20E
  • Combined L-Citrulline and Glutathione Supplementation Increases The

    Combined L-Citrulline and Glutathione Supplementation Increases The

    McKinley-Barnard et al. Journal of the International Society of Sports Nutrition (2015) 12:27 DOI 10.1186/s12970-015-0086-7 RESEARCH Open Access Combined L-citrulline and glutathione supplementation increases the concentration of markers indicative of nitric oxide synthesis Sarah McKinley-Barnard1, Tom Andre1, Masahiko Morita2 and Darryn S. Willoughby1* Abstract Background: Nitric oxide (NO) is endogenously synthesized from L-arginine and L-citrulline. Due to its effects on nitric oxide synthase (NOS), reduced glutathione (GSH) may protect against the oxidative reduction of NO. The present study determined the effectiveness of L-citrulline and/or GSH on markers indicative of NO synthesis in in vivo conditions with rodents and humans and also in an in vitro condition. Methods: In phase one, human umbilical vein endothelial cells (HUVECs) were treated with either 0.3 mM L-citrulline, 1 mM GSH (Setria®) or a combination of each at 0.3 mM. In phase two, Sprague–Dawley rats (8 weeks old) were randomly assigned to 3 groups and received either purified water, L-citrulline (500 mg/kg/day), or a combination of L-citrulline (500 mg/kg/day) and GSH (50 mg/kg/day) by oral gavage for 3 days. Blood samples were collected and plasma NOx (nitrite + nitrate) assessed. In phase three, resistance-trained males were randomly assigned to orally ingest either cellulose placebo (2.52 g/day), L-citrulline (2 g/day), GSH (1 g/day), or L-citrulline (2 g/day) + GSH (200 mg/day) for 7 days, and then perform a resistance exercise session involving 3 sets of 10-RM involving the elbow flexors.
  • The Responses of Glutathione and Antioxidant Enzymes to Hyperoxia in Developing Lung

    The Responses of Glutathione and Antioxidant Enzymes to Hyperoxia in Developing Lung

    LUNG GLUTATHIONE RESPONSE TO HYPEROXIA 8 19 Physiol 55: 1849- 1853 alkalosis on cerebral blood flow in cats. Stroke 5:324-329 21. Lou HC, Lassen NA. Fnis-Hansen B 1978 Decreased cerebral blood flow after 24. Arvidsson S, Haggendal E, Winso 1 1981 Influence on cerebral blood flow of administration of sodium bicarbonate in the distressed newborn infant. Acta infusions of sodium bicarbonate during respiratory acidosis and alkalosis in Neurol Scand 57:239-247 the dog. Acta Anesthesiol Scand 25:146-I52 22. Rapoport SI 1970 Effect ofconcentrated solutions on blood-brain barrier. Am 25. Pannier JL, Weyne J, Demeester G, Leusen 1 1978 Effects of non-respiratory J Physiol 219270-274 alkalosis on brain tissue and cerebral blood flow in rats with damaged blood- 23. Pannier JL, Demeester MS, Leuscn 1 1974 Thc influence of nonrcspiratory brain hamer. Stroke 9:354-359 003 1-3998/85/1908-08 19$0:.00/0 PEDIATRIC RESEARCH Vol. 19, No. 8, 1985 Copyright 8 1985 International Pediatric Research Foundation, Inc Prinled in U.S.A. The Responses of Glutathione and Antioxidant Enzymes to Hyperoxia in Developing Lung JOSEPH B. WARSHAW, CHARLIE W. WILSON, 111, KOTARO SAITO, AND RUSSELL A. PROUGH Departmmls qfP~diufricsarid Biochemistr,~, The University of Texas Health Srience Center ul DaNas. Dallas, Texas 75235 ABSTRACT. Total glutathione levels and the activity of Abbreviations enzymes associated with antioxidant protection in neonatal lung are increased in response to hyperoxia. GIutathione SOD, superoxide dismutiase levels in developing rat lung decreased from 24 nmol/mg GSH, reduced glutathione protein on day 19 of gestation to approximately 12 nmol/ GSSG, oxidized glutathione mg protein at birth.
  • Ibotenic Acid

    Ibotenic Acid

    Ibotenic acid Catalog Number I2765 Storage at Room Temperature Product Description Preparation Instructions Molecular Formula: C5H6N2O4 This product is soluble in water (1 mg/ml) with Molecular Weight: 158.1 < 5 min. sonication, yielding a clear, colorless CAS Number: 2552-55-8 solution. Melting point: 151-152 °C1 Synonyms: α-amino-3-hydroxy-5-isoxazoleacetic Storage/Stability acid1 Store the product desiccated at –20 C and it remains active for at least 3 years. This product is the principal toxin found in many mushroom varieties. Cells metabolize this product to References another active derivative, muscimol. Both of these 1. The Merck Index, 11th ed., Entry# 4808. toxins act as excitatory amino acids by mimicking the 2. Collingridge, et al., Excitatory amino acid receptors natural transmitters, glutamic acid and aspartic acid, on in the vertebrate central nervous system. neurons in the central nervous system.2,3 These toxins Pharmacological Review, 40(2), 143 (1989). may also cause selective death of neurons sensitive to these excitatory amino acids.4,5 This product is a potent 3. Johnston, G. A., et al., Spinal interneuron excitation glutamate agonist, which has been used to potentiate by conformationally restricted analogues of L- anesthesia and to inhibit tremor and emesis. glutamic acid. Nature, 248(5451), 804-805 (1974). 4. Gallagher, M., et al., The amygdala central nucleus This product has also been used to suppress enzymatic and appetitive Pavlovian conditioning: lesions activities. When injected into rat brain, it was shown to impair one class of conditioned behavior. J. suppress choline acetyltransferase activity.6 Seven days after injection, enzyme levels had decreased 60%; Neurosci., 10(6), 1906-1911 (1990) after 3 months activity had returned to normal.
  • Serum Metabolite Profiles As Potential Biochemical Markers in Young

    Serum Metabolite Profiles As Potential Biochemical Markers in Young

    www.nature.com/scientificreports OPEN Serum metabolite profles as potential biochemical markers in young adults with community- acquired pneumonia cured by moxifoxacin therapy Bo Zhou1, Bowen Lou2,4, Junhui Liu3* & Jianqing She2,4* Despite the utilization of various biochemical markers and probability calculation algorithms based on clinical studies of community-acquired pneumonia (CAP), more specifc and practical biochemical markers remain to be found for improved diagnosis and prognosis. In this study, we aimed to detect the alteration of metabolite profles, explore the correlation between serum metabolites and infammatory markers, and seek potential biomarkers for young adults with CAP. 13 Eligible young mild CAP patients between the ages of 18 and 30 years old with CURB65 = 0 admitted to the respiratory medical department were enrolled, along with 36 healthy participants as control. Untargeted metabolomics profling was performed and metabolites including alcohols, amino acids, carbohydrates, fatty acids, etc. were detected. A total of 227 serum metabolites were detected. L-Alanine, 2-Hydroxybutyric acid, Methylcysteine, L-Phenylalanine, Aminoadipic acid, L-Tryptophan, Rhamnose, Palmitoleic acid, Decanoylcarnitine, 2-Hydroxy-3-methylbutyric acid and Oxoglutaric acid were found to be signifcantly altered, which were enriched mainly in propanoate and tryptophan metabolism, as well as antibiotic-associated pathways. Aminoadipic acid was found to be signifcantly correlated with CRP levels and 2-Hydroxy-3-methylbutyric acid and Palmitoleic acid with PCT levels. The top 3 metabolites of diagnostic values are 2-Hydroxybutyric acid(AUC = 0.90), Methylcysteine(AUC = 0.85), and L-Alanine(AUC = 0.84). The AUC for CRP and PCT are 0.93 and 0.91 respectively.