title: Riboflavin and ultraviolet illumination affects selected platelet mRNA transcript amounts differently.
abstract: Pathogen inactivation (PI) techniques use ultraviolet (UV) illumination with or without a photosensitizer to destroy pathogen RNA and DNA. Although lacking a nucleus and innate DNA transcription, platelets (PLTs) contain RNA and can synthesize proteins. The impact of PI on PLT protein synthesis and function is unknown; altered synthesis may affect overall PLT quality. In this study we determine to what extent PLT RNA is affected by PI.
  
  
  
  In a pool-and-split design, paired apheresis PLT concentrates were treated with riboflavin and UV illumination or were left untreated. PLT total RNA and mRNA amounts specific for glycoproteins (GP)IIIa, GPIIb, and GPIb; α-granule proteins PLT factor (PF)4; osteonectin and thrombospondin (TSP); and housekeeping protein glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were determined using absorbance and quantitative polymerase chain reaction.
  
  
  
  After treatment, amounts of all analyzed mRNAs were significantly reduced (p < 0.05), but to different degrees. For GAPDH and PF4, transcripts appeared less susceptible to the treatment, with 70% remaining 1 hour after UV illumination. For GPIIIa and TSP, less than 15% remained after treatment. There was a correlation (R(2)  = 0.85) between transcript length and amount of mRNA remaining 1 hour after treatment. Total RNA demonstrated a life span equal to the PLT life span of 10 to 11 days.
  
  
  
  This is the first report of the impact of riboflavin and UV illumination on PLT mRNA. Results suggest that all mRNA present in PLTs is affected by the treatment although the degree of the effect varies among transcripts.
authors.affiliation: Department of Pathology and Laboratory Medicine and Centre for Blood Research, University of British Columbia.
authors.forename: Christa
authors.initials: C
authors.lastname: Klein-Bosgoed
authors.affiliation: Department of Pathology and Laboratory Medicine and Centre for Blood Research, University of British Columbia.|Canadian Blood Services Centre for Innovation, Vancouver, BC, Canada.
authors.forename: Peter
authors.initials: P
authors.lastname: Schubert
authors.affiliation: Department of Pathology and Laboratory Medicine and Centre for Blood Research, University of British Columbia. dana.devine@blood.ca.|Canadian Blood Services Centre for Innovation, Vancouver, BC, Canada.. dana.devine@blood.ca.
authors.forename: Dana V
authors.initials: DV
authors.lastname: Devine
mesh_terms: D001792:Blood Platelets / Q000187:drug effects / Q000378:metabolism* / Q000528:radiation effects; D001793:Blood Preservation / Q000379:methods; D024581:Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating) / Q000032:analysis / Q000235:genetics; D006801:Humans; D039661:Integrin beta3 / Q000032:analysis / Q000235:genetics; D015676:Osteonectin / Q000032:analysis / Q000235:genetics; D010978:Platelet Factor 4 / Q000032:analysis / Q000235:genetics; D040201:Platelet Membrane Glycoprotein IIb / Q000032:analysis / Q000235:genetics; D012333:RNA, Messenger / Q000187:drug effects / Q000235:genetics* / Q000528:radiation effects; D012256:Riboflavin / Q000494:pharmacology*; D019699:Thrombospondins / Q000032:analysis / Q000235:genetics; D014466:Ultraviolet Rays
chemical_list: D039661:Integrin beta3; D015676:Osteonectin; D040201:Platelet Membrane Glycoprotein IIb; D012333:RNA, Messenger; D019699:Thrombospondins; D010978:Platelet Factor 4; D024581:Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating); D012256:Riboflavin