title: Continuous monitoring of highly reactive oxygen radicals during in vivo microdialysis.
abstract: BACKGROUND
  Terephthalate (TA(2-)), which reacts with highly reactive oxygen species (hROS) to form the fluorophor 2-hydroxy terephthalic acid (OH-TA) with a high selectivity, has been used for determining hROS formation during in vivo microdialysis. Previously this involved collecting fractions of the microdialysate and determining the OH-TA formed after HPLC (the batch method).
  
  
  METHODS
  This work reports the development and validation of a procedure for continuously determining hROS formation during microdialysis. TA(2-) was added to the artificial cerebrospinal fluid (aCSF) perfusing medium to trap hROS. OH-TA formation was detected in real time with a sensitive fluorescence detector equipped with a capillary flow cell that was coupled directly to the effluent stream of the microdialysis system.
  
  
  RESULTS
  The behaviour of the system was assessed by comparison with the batch method and using a well-characterized animal model of excitotoxic damage, based on the application of high concentrations (1mM and 500μM) of the non-NMDA glutamate receptor agonist kainate (KA) to the neostriatum. Data for the evoked release of taurine were also determined in these samples. No temporal difference between hROS and taurine release could be detected.
  
  
  CONCLUSIONS
  The flow method had a comparable sensitivity of hROS detection to the batch method. It was simpler, cheaper and less time-consuming than the batch method.
  
  
  CONCLUSIONS
  This direct system is convenient and technically undemanding. It should be useful for the rapid assessment of the hROS responses to neurotoxins and other compounds in microdialysis experiments in vivo.
authors.affiliation: Institute for Applied Synthetic Chemistry, Vienna University of Technology, Getreidemarkt 9/163-AC, A-1060 Vienna, Austria.
authors.forename: Bashkim
authors.initials: B
authors.lastname: Misini
authors.affiliation: Institute for Applied Synthetic Chemistry, Vienna University of Technology, Getreidemarkt 9/163-AC, A-1060 Vienna, Austria.
authors.forename: Wolfhardt
authors.initials: W
authors.lastname: Freinbichler
authors.affiliation: Dipartimento di Neuroscienze, Psicologia, Area del Farmaco e Salute del Bambino (NEUROFARBA), Università degli Studi di Firenze, Viale G. Pieraccini 6, 50139 Firenze, Italy.
authors.forename: Maria Alessandra
authors.initials: MA
authors.lastname: Colivicchi
authors.affiliation: Department of Biology, University of Prishtina, Kosovo.
authors.forename: Kemajl
authors.initials: K
authors.lastname: Bisilimi
authors.affiliation: Institute for Applied Synthetic Chemistry, Vienna University of Technology, Getreidemarkt 9/163-AC, A-1060 Vienna, Austria.
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Linert
authors.affiliation: School of Biochemistry and Immunology, Trinity College Dublin, Dublin 2, Ireland.
authors.forename: Keith F
authors.initials: KF
authors.lastname: Tipton
authors.affiliation: Dipartimento di Neuroscienze, Psicologia, Area del Farmaco e Salute del Bambino (NEUROFARBA), Università degli Studi di Firenze, Viale G. Pieraccini 6, 50139 Firenze, Italy. Electronic address: laura.dellacorte@unifi.it.
authors.forename: Laura
authors.initials: L
authors.lastname: Della Corte
mesh_terms: D000704:Analysis of Variance; D000818:Animals; D019540:Area Under Curve; D002851:Chromatography, High Pressure Liquid; D004195:Disease Models, Animal; D018690:Excitatory Amino Acid Agonists / Q000633:toxicity; D005453:Fluorescence; D007608:Kainic Acid / Q000633:toxicity; D008297:Male; D017551:Microdialysis / Q000379:methods*; D017072:Neostriatum / Q000378:metabolism*; D020258:Neurotoxicity Syndromes / Q000209:etiology / Q000378:metabolism* / Q000473:pathology; D009862:Online Systems; D010795:Phthalic Acids / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D017382:Reactive Oxygen Species / Q000032:analysis / Q000378:metabolism*; D013654:Taurine / Q000378:metabolism; D013997:Time Factors
chemical_list: D018690:Excitatory Amino Acid Agonists; D010795:Phthalic Acids; D017382:Reactive Oxygen Species; D013654:Taurine; C011363:terephthalic acid; D007608:Kainic Acid
keywords: Capillary flow cell; Fluorescence recording; Highly reactive oxygen species (hROS); Online in vivo monitoring; Terephthalate