Abstract
Maple syrup urine disease is an autosomal metabolic disease caused by a deficiency of branched-chain α-keto acid dehydrogenase complex activity. In this disease occur the accumulation of the branched-chain amino acids leucine, isoleucine, and valine and their corresponding branched-chain α-keto acids in the tissues and body fluids. The affected patients may present psychomotor development delay and mental retardation. The pathophysiology of maple syrup urine disease is not entirely understood, but leucine seems to be the primary neurotoxic metabolite. Creatine and pyruvate are energetics and antioxidants substances. In this study, we investigated the effects of leucine administration and co-administration of creatine plus pyruvate on several parameters of oxidative stress and phosphoryl transfer network in cerebral cortex and hippocampus of Wistar rats treated from the 8th to the 21st postpartum day. Leucine induced oxidative stress and diminished the activities of pyruvate kinase, adenylate kinase, cytosolic and mitochondrial creatine kinase. Co-administration of creatine plus pyruvate prevented the alterations provoked by leucine administration on the oxidative stress and the enzymes of phosphoryltransfer network. These results indicate that chronic administration of leucine may stimulate oxidative stress and alters the enzymes of phosphoryltransfer network in the cerebral cortex and hippocampus of the rats. It is possible that these effects may contribute, along with other mechanisms, to the neurological dysfunction found in patients affected by maple syrup urine disease. In this case, it is possible that creatine plus pyruvate supplementation could benefit to the patients.
Similar content being viewed by others
References
Aksenov MY, Markesbery WR (2001) Changes in thiol content and expression of glutathione redox system genes in the hippocampus and cerebellum in Alzheimer’s disease. Neurosci Lett 302:41–145
Aksenov M, Aksenova M, Butterfield AD, Markesbery WR (2000) Oxidative modification of creatine kinase BB in Alzheimer’s disease brain. J Neurochem 74:2520–2527
Amaral AU, Leipnitz G, Fernandes CG, Seminotti B, Schuck PF, Wajner M (2010) Alpha-ketoisocaproic acid and leucine provoke mitochondrial bioenergetic dysfunction in rat brain. Brain Res 1324:75–84
Andrade VS, Rojas DB, Oliveira L, Nunes ML, de Castro FL, Garcia C, Gemelli T, de Andrade RB, Wannmacher CM (2012) Creatine and pyruvate prevent behavioral and oxidative stress alterations caused by hypertryptophanemia in rats. Mol Cell Biochem 362:225–232
de Andrade RB, Gemelli T, Rojas DB, Funchal C, Dutra-Filho CS, Wannmacher CM (2012) Tyrosine impairs enzymes of energy metabolism in cerebral cortex of rats. Mol Cell Biochem 364:253–261
de Andrade RB, Gemelli T, Rojas DB, Bonorino NF, Costa B, Funchal C, Dutra-Filho CS, Wannmacher CM (2015) Creatine and pyruvate prevent the alterations caused by tyrosine on parameters of oxidative stress and enzymes activities of phosphoryltransfer network in cerebral cortex of wistar rats. Mol Neurobiol 51:1184–1194
Andrae U, Singh J, Ziegler-Skylakakis K (1985) Pyruvate and related a-ketoacids protect mammalian cells in culture against hydrogen peroxide-induced cytotoxicity. Toxicol let 28:93–98
Andres RH, Ducray AD, Schlattner U, Wallimann T, Widmer HR (2008) Functions and effects of creatine in the central nervous system. Brain Res Bull 76:329–343
Araki T, Sasaki Y, Milbrandt J (2004) Increased nuclear NAD biosynthesis and SIRT1 activation prevent axonal degeneration. Science 305:1010–1013
Artuch R, Colomé C, Sierra C, Brandi N, Lambruschini J, Urgate D, Vilaseca MA (2004) A longitudinal study of antioxidant status in Phenylketonuric patients. Clin Biochem 37:198–203
Barschak AG, Sitta A, Deon M, de Oliveira MH, Haeser A, Dutra-Filho CS, Wajner M, Vargas CR (2006) Evidence that oxidative stress is increased in plasma from patients with maple syrup urine disease. Metab Brain Dis 21:279–286
Barschak AG, Marchesan C, Sitta A, Deon M, Giugliani R, Wajner M, Vargas CR (2008a) Maple syrup urine disease in treated patients: biochemical and oxidative stress profiles. Clin Biochem 41:317–324
Barschak AG, Sitta A, Deon M, Barden AT, Dutra-Filho CS, Wajner M, Vargas CR (2008b) Oxidative stress in plasma from maple syrup urine disease patients during treatment. Metab Brain Dis 23:71–80
Barschak AG, Sitta A, Deon M, Busanello EN, Coelho DM, Cipriani F, Dutra-Filho CS, Giugliani R, Wajner M, Vargas CR (2009) Amino acids levels and lipid peroxidation in maple syrup urine disease patients. Clin Biochem 42:462–466
Beal MF (1995) Aging, energy and oxidative stress in neurodegenerative diseases. Ann Neurol 38:357–366
Beal MF (2000) Energetics in the pathogenesis of neurodegenerative diseases. Trends Neurosci 23:298–304
Beal M (2011) Neuroprotective effects of creatine. Amino Acids 40:1305–1313
Bender A, Koch W, Elstner M, Schombacher Y, Bender J, Moeschl M, Gekeler F, Müller-Myhsok B, Gasser T, Tatsch K, Klopstock T (2006) Creatine supplementation in Parkinson disease: a placebo controlled randomized pilot trial. Neurology 67:1262–1264
Bender A, Beckers J, Schneider I, Hölter SM, Haack T, Ruthsatz T, Vogt-Weisenhorn DM, Becker L, Genius J, Rujescu D, Irmler M, Mijalski T, Mader M, Quintanilla-Martinez L, Fuchs H, Gailus-Durner V, de Angelis MH, Wurst W, Schmidt J, Klopstock T (2008) Creatine improves health and survival of mice. Neurobiol Aging 29:1404–1411
Berti SL, Nasi GM, Garcia C, Castro FL, Nunes ML, Rojas DB, Moraes TB, Dutra-Filho CS, Wannmacher CM (2012) Pyruvate and creatine prevent oxidative stress and behavioral alterations caused by phenylalanine administration into hippocampus of rats. Metab Brain Dis 27:79–89
Brewer GJ, Wallimann TW (2000) Protective effect of the energy precursor creatine against toxicity of glutamate and betaamyloid in rat hippocampal neurons. J Neurochem 74:1968–1978
Bridi R, Araldi J, Sgarbi MB, Testa CG, Durigon K, Wajner M, Dutra-Filho CS (2003) Induction of oxidative stress in rat brain by the metabolites accumulating in maple syrup urine disease. Int J Devl Neuroscience 21:327–332
Bridi R, Latini A, Braun CA, Zorzi GK, Wajner M, Lissi EG, Dutra-Filho CS (2005) Evaluation of the mechanisms involved in leucine induced oxidative damage in cerebral cortex of young rats. Free Radic res 39:71–79
Burlacu A, Jinga V, Gafencu AV, Simionescu M (2001) Severity of oxidative stress generates different mechanisms of endothelial cell death. Cell Tissue Res 306:409–416
Change B, Sies H, Boveris A (1979) Hydroperoxide metabolism in mammalian organs. Physiol Rev 59:527–605
Chaturvedi RK, Beal MF (2008) Mitochondrial approaches for neuroprotection. Ann N Y Acad Sci. 1147:395–412
Chuang DT, Shih VE (2001) Maple syrup urine disease (branched-chain ketoaciduria). In: Scriver CR, Beaudet AL, Sly WS, Valle D (eds) The metabolic & molecular bases of inherited diseases, 8th edn. McGraw-Hill, New York, pp 1667–1724
Cornelio AR, Rodrigues V Jr, de Souza Wyse AT, Dutra-Filho CS, Wajner M, Wannmacher CM (2004) Tryptophan reduces creatine kinase activity in the brain cortex of rats. Int J Dev Neurosci 22:95–101
Costabeber E, Kessler A, Severo Dutra-Filho C, de Souza Wyse AT, Wajner M, Wannmacher CM (2003) Hyperphenylalaninemia reduces creatine kinase activity in the cerebral cortex of rats. Int J Dev Neurosci 21:111–116
Das UN (2006) Pyruvate is an endogenous anti-inflammatory and anti-oxidant molecule. Med Sci Monit 12:79–84
Desagher S, Glowinski J, Premont J (1997) Pyruvate protects neurons against hydrogen peroxide-induced toxicity. J Neurosci 17:9060–9067
Dos Reis EA, Rieger E, de Souza SS, Rasia-Filho AA, Wannmacher CM (2013) Effects of a co-treatment with pyruvate and creatine on dendritic spines in rat hippocampus and posterodorsal medial amygdala in a phenylketonuria animal model. Metab Brain dis 28:509–517
Dzeja PP, Terzic A (1998) Phosphotransfer reactions in the regulation of ATP-sensitive K+ channels. FASEB j 12:523–529
Dzeja PP, Terzic A (2003) Phosphotransfer networks and cellular energetics. J Exp Biol 206:2039–2047
Dzeja PP, Vitkevicius KT, Redfied MM, Burnett JC, Terzic A (1999) Adenylate kinase-catalyzed phosphotransfer in the myocardium: increased contribution in heart failure. Circ Res 84:1137–1143
Dzeja PP, Redfield MM, Burnett JC, Terzic A (2000) Failing energetics in failing heats. Curr Cardiol rep 2:212–217
Esterbauer H, Cheeseman KH (1990) Determination of aldehydic lipid peroxidation products: malonaldehyde and 4-hydroxynonenal. Method Enzymol 186:407–421
Feksa LR, Cornelio AR, Dutra-Filho CS, de Souza Wyse AT, Wajner M, Wannmacher CM (2003) Characterization of the inhibition of pyruvate kinase caused by phenylalanine and phenylpyruvate in rat brain cortex. Brain Res 968:199–205
Feksa LR, Cornelio A, Dutra-Filho CS, Wyse ATS, Wajner M, Wannmacher CM (2004) Inhibition of pyruvate kinase activity by cystine in brain cortex of rats. Brain Res 1012:93–100
Feksa LR, Dutra-Filho CS, Wyse ATS, Wajner M, Wannmacher CMD (2005) The effects of the interactions between amino acids on pyruvate kinase activity from the brain cortex of young rats. Int J Dev Neurosci 23:509–514
Feksa LR, Latini A, Rech VC, Feksa PB, Koch GD, Amaral MF, Leipnitz G, Dutra-Filho CS, Wajner M, Wannmacher CM (2008) Tryptophan administration induces oxidative stress in brain cortex of rats. Metab Brain Dis 23:221–233
Ferrante RJ, Andreassen OA, Jenkins BG, Dedeoglu A, Kuemmerle S, Kubilus JK, Kaddurah-Daouk R, Hersch SM, Beal MF (2000) Neuroprotective effects of creatine in a transgenic mouse model of Huntington’s disease. J Neurosci 20:4389–4397
Figueiredo VC, Feksa LR, Wannmacher CM (2009) Cysteamine prevents inhibition of adenylate kinase caused by cysteine in rat brain cortex. Metab Brain Dis 24:373–381
Fleck RMM, Rodrigues-Junior V, Giacomazzi J, Parissoto D, Dutra-Filho CS, Wyse ATS, Wajner M, Wannmacher CMD (2005) Cysteamine prevents and reverses the inhibition of creatine kinase activity caused by cystine in rat brain cortex. Neurochem Int 46:391–397
de Franceschi ID, Rieger E, Vargas AP, Rojas DB, Campos AG, Rech VC, Feksa LR, Wannmacher CM (2013) Effect of leucine administration to female rats during pregnancy and lactation on oxidative stress and enzymes activities of phosphoryltransfer network in cerebral cortex and hippocampus of the offspring. Neurochem Res 38:632–643
Gemelli T, de Andrade RB, Rojas DB, Bonorino NF, Mazzola PN, Tortorelli LS, Funchal C, Dutra-Filho CS, Wannmacher CM (2013) Effect of β-alanine administration on selected parameters of oxidative stress and phosphoryltransfer network in cerebral cortex and cerebellum of rats. Mol Cell Biochem 380:161–170
Gilbert HF (1984) Redox control of enzyme activities by thiol/disulfide exchange. Method Enzymol 107:330–351
Guerreiro G, Mescka CP, Sitta A, Donida B, Marchetti D, Hammerschmidt T, Faverzani J, Coelho DM, Wajner M, Dutra-Filho CS, Vargas CR (2015) Urinary biomarkers of oxidative damage in maple syrup urine disease: the L-carnitine role. Int J Dev Neurosci 42:10–14
Halliwell B (2001) Role of free radicals in the neurodegenerative diseases: therapeutic implications for antioxidant treatment. Drugs Aging 18:685–716
Halliwell B (2006) Oxidative stress and neurodegeneration: where are we now? J Neurochem 97:1634–1658
Halliwell B (2013) The antioxidant paradox: less paradoxical now? Br J Clin Pharmacol 75:637–644
Halliwell B, Gutteridge JMC (1996) Oxygen radicals and nervous system. Trends Neurosci 8:22–26
Halliwell B, Gutteridge JMC (2007) Free radical in biology and medicine. In: Cellular responses to oxidative stress: adaptation, damage, repair, senescence and death, 4th edn. Oxford University Press, Oxford, pp 188–276
Halliwell B, Gutteridge JMC (2015) Free radicals in biology and medicine, Fifth edn. Oxford University Press, Oxford
Hayashi M, Miyata R, Tanuma N (2012) Oxidative stress in developmental brain disorders. Adv Exp Med Biol 724:278–290
Holtzman D, Togliatti A, Khait I, Jensen F (1998) Creatine increases survival and suppresses seizures in the hypoxic immature rat. Pediatr Res 44:410–414
Hughes BP (1962) A method for estimation of serum creatine kinase and its use in comparing creatine kinase and aldolase activity in normal and pathological sera. Clin Chim Acta 7:597–603
Jagtap JC, Chandele A, Chopde BA, Shastry P (2003) Sodium pyruvate protects against H2O2 mediated apoptosis in human neuroblastoma cell line-SK-N-MC. J Chem Neuroanat 26:109–118
Kehrer JP (2000) The Haber-Weiss reaction and mechanisms of toxicity. Toxicology 149:43–50
Kessler A, Costabeber E, Dutra-Filho CS, Wyse AT, Wajner M, Wannmacher CM (2003) Proline reduces creatine kinase activity in the brain cortex of rats. Neurochem Res 28:1175–1180
Kim JB, Yu YM, Kim SW, Lee JK (2005) Anti-inflammatory mechanism is involved in ethyl pyruvate-mediated efficacious neuroprotection in the postischemic brain. Brain Res 1060:188–192
Klein AM, Ferrante RJ (2007) The neuroprotective role of creatine. Subcell Biochem 46:205–243
Koga Y, Povalko N, Katayama K, Kakimoto N, Matsuishi T, Naito E, Tanaka M (2012) Beneficial effect of pyruvate therapy on Leigh syndrome due to a novel mutation in PDH E1α gene. Brain Dev 34:87–91
Lawler JM, Barnes WS, Wu G, Song W, Demaree S (2002) Direct antioxidant properties of creatine. Biochem Biophys Res Commun 290:47–52
LeBel CP, Ischiropoulos H, Bondy SC (1992) Evaluation of the probe 20,70-dichlorofluorescin as an indicator of reactive oxygen species formation and oxidative stress. Chem Res Toxicol 5:227–231
Leong SF, Lai JCK, Lim L, Clark JB (1981) Energy-metabolising enzymes in brain regions of adult and aging rats. J Neurochem 37:1548–1556
Lowry OH, Rosebrough N, Farr AL, Randal RJ (1951) Protein measurementwith a Folin phenol reagent. J Biol Chem 193:265–275
Lu SC (2009) Regulation of glutathione synthesis. Mol Asp Med 30:42–59
Macedo LGRP, Carvalho-Silva M, Ferreira GK, Vieira JS, Olegário N, Gonçalves RC, Vuolo FS, Ferreira GC, Schuck PF, Dal-Pizzol F, Streck EL (2013) Effect of acute administration of L-tyrosine on oxidative stress parameters in brain of young rats. Neurochem Res 38:2625–2630
Marklund SL (1985) Pyrogallol autoxidation. In: Greenwald RA (ed) Handbook of methods for oxygen radical research. CRC Press Inc, Boca Raton, pp 243–247
Matthews RT, Yang L, Jenkins BG, Ferrante RJ, Rosen BR, Kaddurah-Daouk R, Beal MF (1998) Neuroprotective effects of creatine and cyclocreatine in animal models of Huntington’s disease. J Neurosci 18:156–163
Mazzio E, Soliman KFA (2003a) Pyruvic acid cytoprotection against 1-methyl-4-phenylpyridinium, 6-hydroxydopamine and hydrogen peroxide toxicities in vitro. Neurosci Lett 337:77–80
Mazzio EA, Soliman KF (2003b) Cytoprotection of pyruvic acid and reduced beta-nicotinamide adenine dinucleotide against hydrogen peroxide toxicity in neuroblastoma cells. Neurochem Res 28:733–741
Mescka C, Moraes T, Rosa A, Mazzola P, Piccoli B, Jacques C, Dalazen G, Coelho J, Cortes M, Terra M, Regla Vargas C, Dutra-Filho CS (2011) In vivo neuroprotective effect of L-carnitine against oxidative stress in maple syrup urine disease. Metab Brain Dis 26:21–28
Mescka CP, Wayhs CA, Vanzin CS (2013) Protein and lipid damage in maple syrup urine disease patients: l-carnitine effect. Int J Dev Neurosci 31:21–24
Mescka CP, Wayhs CA, Guerreiro G, Manfredini V, Dutra-Filho CS, Vargas CR (2014) Prevention of DNA damage by L-carnitine induced by metabolites accumulated in maple syrup urine disease in human peripheral leukocytes in vitro. Gene 548:294–298
Mochel F, Durant B, Meng X, O’Callaghan J, Yu H, Brouillet E, Wheeler VC, Humbert S, Schiffmann R, Durr A (2012) Early alterations of brain cellular energy homeostasis in huntington disease models. J Biol Chem 287:1361–1370
Mukherjee SK, Klaidman LK, Yasharel R, Adams JD (1997) Increased brain NAD prevents neuronal apoptosis in vivo. Eur J Pharmacol 330:27–34
O’Donnell-Tormey J, Nathan CF, Lanks K, DeBoer CJ, de la Harpe J (1987) Secretion of pyruvate. An antioxidant defense of mammalian cells. J Exp Med 165:500–514
Ohkawa H, Ohishi N, Yagi K (1979) Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem 95:351–358
Owen L, Sunram-Lea SI (2011) Metabolic agents that enhance ATP can improve cognitive functioning: a review of the evidence for glucose, oxygen, pyruvate, creatine, and l-carnitina. Nutrients 3:735–755
Park MA, Thoene JG (2005) Potential role of apoptosis in development of the cystinotic phenotype. Pediatr Nephrol 20:441–446
Persky AM, Brazeau GA (2001) Clinical pharmacology of the dietary supplement creatine monohydrate. Pharmacol Rev 53:161–176
Pilla C, Cardozo RFO, Dutra-Filho CS, Wyse ATS, Wajner M, Wannmacher CM (2003) Effect of leucine administration on creatine kinase activity in rat brain. Metab Brain Dis 18:17–25
Price NC, Cohn M, Schirmer RH (1975) Fluorescent and spin label probes of the environments of the sulfhydryl groups of porcine muscle adenylate kinase. J Biol Chem 250:644–652
Reed TT (2011) Lipid peroxidation and neurodegenerative disease. Free Radic Biol med 51:1302–1319
Ryu JK, Choi HB, Mclarnon JB (2006) Combined minocycline plus pyruvate treatment enhances effects of each agent to inhibit inflammation, oxidative damage, and neuronal loss in an excitotoxic animal model of Huntington’s disease. Neurosci 141:1835–1848
Saks V, Dzeja P, Schlattner U, Vendelin M, Terzic A, Wallimann T (2006) Cardiac system bioenergetics: metabolic basis of the Frank-Starling law. J Physiol 571:253–273
Sartini S, Sestili P, Colombo E, Martinelli C, Bartolini F, Ciuffoli S, Lattanzi D, Sisti D, Cuppini R (2012) Creatine affects in vitro electrophysiological maturation of neuroblasts and protects them from oxidative stress. J Neurosci Res 90:435–446
Schönberger S, Schweiger B, Schwahn B, Schwarz M, Wendel U (2004) Dysmyelination in the brain of adolescents and young adults with maple syrup urine disease. Mol Genet Metab 82:69–75
Sestili P, Martinelli C, Bravi G, Piccoli G, Curci R, Battistell M, Falcieri E, Agostini D, Gioacchini AM, Stocchi V (2006) Creatine supplementation affords cytoprotection in oxidatively injured cultured mammalian cells via direct antioxidant activity. Free Radic Biol med 40:837–849
Skvorak KJ (2009) Animal models of maple syrup urine disease. J Inherit Metab Dis 32:229–246
Snyderman SE, Norton PM, Roitman E (1964) Maple syrup urine disease with particular reference to diet terapy. Pediatrics 34:454–472
Stadtman ER, Levine RL (2003) Free-radical mediated oxidation of free amino acids and amino acid residues in proteins. Amino Acids 25:207–218
Stockler S, Holzbach U, Hanenfeld F, Marquardt I, Helms G, Requart M, Hanicke W, Frahm J (1994) Creatine deficiency in the brain: a new, treatable inborn error of metabolism. Pediatric res 36:409–413
Strauss KA, Wardley B, Robinson D, Hendrickson C, Rider NL, Puffenberger EG, Shelmer D, Moser AB, Morton DH (2010) Classical maple syrup urine disease and brain development: principles of management and formula design. Mol Genet Metab 99:333–345
Tarnopolsky MA (2007) Clinical use of creatine in neuromuscular and neurometabolic disorders. Subcell Biochem 46:183–204
Tomimoto H, Yamamoto K, Homburger HA, Yanagihara T (1993) Immunoelectron microscopic investigation of creatine kinase BB isoenzyme after cerebral ischemia in gerbils. Acta Neuropathol 86:447–455
Valentini G, Chiarelli LR, Fortin R, Speranza ML, Galizzi A, Mattevi A (2000) The allosteric regulation of pyruvate kinase. J Biol Chem 275:18145–18152
Vlassenko AG, Rundle MM, Raichle ME, Mintun MA (2006) Regulation of blood flow in activated human brain by cytosolic NADH/NAD ratio. Proc Natl Acad Sci U S a 103:1964–1969
Wallimann T, Wyss M, Brdiczka D, Nicolay K, Eppenberger HM (1992) Intracellular compartmentation, structure and function of creatine kinase isoenzymes in tissues with high and fluctuating anergy demands: the ‘phosphocreatine cicuit’ for cellular energy homeostasis. Biochem J 281:21–40
Wendel A (1981) Glutathione peroxidase. Methods Enzymol 77:325–332
Wendt S, Schlattner U, Wallimann T (2003) Differential effects of peroxynitrite on human mitochondrial creatine kinase isoenzymes. Inactivation, octamer destabilization, and identification of involved residues. J Biol Chem 278:1125–30
Wyss M, Kaddurah-Daouk R (2000) Creatine and creatinine metabolism. Physiol rev 80:1107–1213
Yudkoff M (1997) Brain metabolism of branched-chain amino acids. Glia 21:92–98
Zeng J, Yang GY, Ying W, Kelly M, Hira K, James TL, Swanson RA, Litt L (2007) Pyruvate improves recovery after PARP-1-associated energy failure induced by oxidative stress in neonatal rat cerebro cortical slices. J Cereb Blood Flow Metab 27:304–315
Zilberter M, Ivanov A, Ziyatdinova S, Mukhtarov M, Malkov A, Alpár A, Tortoriello G, Botting CH, Fülöp L, Osypov AA, Pitkänen A, Tanila H, Harkany T, Zilberter Y (2013) Dietary energy substrates reverse early neuronal hyperactivity in a mouse model of Alzheimer’s disease. J Neurochem 125:157–171
Acknowledgments
This work was supported by the research grants from, Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) number 306928/2014-0, Fundação de Amparo a Pesquisa do Estado do Rio Grande do Sul (FAPERGS).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Rieger, E., de Franceschi, I.D., Preissler, T. et al. Neuroprotective Effect of Creatine and Pyruvate on Enzyme Activities of Phosphoryl Transfer Network and Oxidative Stress Alterations Caused by Leucine Administration in Wistar Rats. Neurotox Res 32, 575–584 (2017). https://doi.org/10.1007/s12640-017-9762-5
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12640-017-9762-5