title: Evaluation of the ability of aqueous black walnut extracts to induce the production of reactive oxygen species.
abstract: OBJECTIVE
  To assess the in vitro capability of aqueous black walnut extracts (BWEs) to generate reactive oxygen species in water-based media ranging in makeup from a simple buffer solution to a complex solution containing serum.
  
  
  METHODS
  3 BWEs.
  
  
  METHODS
  Production of reactive oxygen species by BWEs prepared in water or N-hexane was tested in PBS solution, PBS solution containing 0.5% bovine serum albumin and 5mM glucose (PBG), and RPMI-1640 medium (RPMI) containing 10% fetal bovine serum or 10% donor horse serum. Reactive oxygen species production was measured as conversion of nonfluorescent dihydrorhodamine 123 by reactive oxygen species to its fluorescent product, rhodamine-123. Hydrogen peroxide was used as a standard for reactive oxygen species activity.
  
  
  RESULTS
  BWEs prepared in water generated reactive oxygen species in a dose-dependent manner over a 4-hour period, with peak activity detected when the BWEs were added as 10% (vol/vol) of the RPMI. The BWE prepared in N-hexane generated maximal reactive oxygen species activity after incubation for 3 to 4 hours when added at concentrations ranging from 0.3% to 0.5% (vol/vol) of the RPMI. The BWE prepared in water generated the highest fluorescent signal in PBS solution, whereas the BWE prepared in N-hexane generated the highest fluorescent signal in PBG.
  
  
  CONCLUSIONS
  The BWEs prepared in water generated a dose-dependent induction of fluorescence in all the water-based solutions tested. These findings indicated that the BWEs, which are used to induce laminitis in horses, generate reactive oxygen species.
authors.affiliation: Food Animal Health and Management Program, Department of Population Health, College of Veterinary Medicine, University of Georgia, Athens, GA 30602, USA. djhurley@uga.edu
authors.forename: David J
authors.initials: DJ
authors.lastname: Hurley
authors.forename: Katherine A E
authors.initials: KA
authors.lastname: Hurley
authors.forename: Kimberly L
authors.initials: KL
authors.lastname: Galland
authors.forename: Bradley
authors.initials: B
authors.lastname: Baker
authors.forename: Londa J
authors.initials: LJ
authors.lastname: Berghaus
authors.forename: James N
authors.initials: JN
authors.lastname: Moore
authors.forename: Rita S K
authors.initials: RS
authors.lastname: Majerle
mesh_terms: D000818:Animals; D005534:Foot Diseases / Q000276:immunology / Q000662:veterinary*; D006734:Horse Diseases / Q000209:etiology* / Q000276:immunology; D006736:Horses; D031324:Juglans / Q000737:chemistry*; D008070:Lipopolysaccharides / Q000378:metabolism; D010936:Plant Extracts / Q000737:chemistry*; D017382:Reactive Oxygen Species / Q000378:metabolism*
chemical_list: D008070:Lipopolysaccharides; D010936:Plant Extracts; D017382:Reactive Oxygen Species