title: Cold storage of liver microorgans in ViaSpan and BG35 solutions: study of ammonia metabolism during normothermic reoxygenation.
abstract: BACKGROUND
  This work focuses on ammonia metabolism of Liver Microorgans (LMOs) after cold preservation in a normothermic reoxygenation system (NRS). We have previously reported the development of a novel preservation solution, Bes-Gluconate-PEG 35 kDa (BG35) that showed the same efficacy as ViaSpan to protect LMOs against cold preservation injury. The objective of this work was to study mRNA levels and activities of two key Urea Cycle enzymes, Carbamyl Phosphate Synthetase I (CPSI) and Ornithine Transcarbamylase (OTC), after preservation of LMOs in BG35 and ViaSpan and the ability of these tissue slices to detoxify an ammonia overload in a NRS model.
  
  
  METHODS
  After 48 h of cold storage (0°C in BG35 or ViaSpan) LMOs were rewarmed in KHR containing an ammonium chloride overload (1 mM). We determined ammonium detoxification capacity (ADC), urea synthesis and enzyme activities and relative mRNA levels for CPSI and OTC.
  
  
  RESULTS
  At the end of reoxygenation LMOs cold preserved in BG35 have ADC and urea synthesis similar to controls. ViaSpan group demonstrated a lower capacity to detoxify ammonia and to synthesize urea than fresh LMOs during the whole reoxygenation period which correlated with the lower mRNA levels and activities for CPSI and OTC observed for this group.
  
  
  CONCLUSIONS
  We demonstrate that our preservation conditions (48 hours, BG35 solution, anoxia, 0ºC) did not affect ammonia metabolism of cold preserved LMOs maintaining the physiological and biochemical liver functions tested, which allows their future use as biological component of a BAL system.
authors.affiliation: Centro Binacional de Criobiología Clínica y Aplicada (CAIC), Universidad Nacional de Rosario, Arijón 28 bis, (2000) Rosario, Argentina.
authors.forename: María Dolores
authors.initials: MD
authors.lastname: Pizarro
authors.affiliation: Centro Binacional de Criobiología Clínica y Aplicada (CAIC), Universidad Nacional de Rosario, Arijón 28 bis, (2000) Rosario, Argentina.
authors.forename: María Gabriela
authors.initials: MG
authors.lastname: Mediavilla
authors.affiliation: Centro Binacional de Criobiología Clínica y Aplicada (CAIC), Universidad Nacional de Rosario, Arijón 28 bis, (2000) Rosario, Argentina.
authors.forename: Florencia
authors.initials: F
authors.lastname: Berardi
authors.affiliation: Centro Studi Fegato, AREA Science Park, Basovizza SS 14, Km 163,5. 34012 Trieste and Department of Medical Sciences, University of Trieste, Italy.
authors.forename: Claudio
authors.initials: C
authors.lastname: Tiribelli
authors.affiliation: Centro Binacional de Criobiología Clínica y Aplicada (CAIC), Universidad Nacional de Rosario, Arijón 28 bis, (2000) Rosario, Argentina.
authors.forename: Joaquín V
authors.initials: JV
authors.lastname: Rodríguez
authors.affiliation: Farmacología, Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, Suipacha 531, S2002 LRK Rosario, Argentina.
authors.forename: María Eugenia
authors.initials: ME
authors.lastname: Mamprin
mesh_terms: D000241:Adenosine / Q000494:pharmacology; D000493:Allopurinol / Q000494:pharmacology; D000641:Ammonia / Q000378:metabolism*; D000818:Animals; D002222:Carbamoyl-Phosphate Synthase (Ammonia) / Q000378:metabolism; D003080:Cold Temperature; D005978:Glutathione / Q000494:pharmacology; D007328:Insulin / Q000494:pharmacology; D008099:Liver / Q000187:drug effects* / Q000378:metabolism* / Q000473:pathology; D008111:Liver Function Tests; D008297:Male; D023421:Models, Animal; D009926:Organ Preservation / Q000379:methods*; D019188:Organ Preservation Solutions / Q000494:pharmacology*; D009954:Ornithine Carbamoyltransferase / Q000378:metabolism; D010100:Oxygen / Q000008:administration & dosage / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D011887:Raffinose / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D015424:Reperfusion; D013997:Time Factors
chemical_list: D007328:Insulin; D019188:Organ Preservation Solutions; D012333:RNA, Messenger; C055206:University of Wisconsin-lactobionate solution; D000493:Allopurinol; D000641:Ammonia; D009954:Ornithine Carbamoyltransferase; D002222:Carbamoyl-Phosphate Synthase (Ammonia); D005978:Glutathione; D000241:Adenosine; D011887:Raffinose; D010100:Oxygen