title: Periods without agitation diminish platelet mitochondrial function during storage.
abstract: BACKGROUND
  Prolonged periods without agitation produce platelet (PLT) storage lesions that result in reduced in vitro assay parameters and an increase of apoptotic markers during storage. The aim of this study was to evaluate the influence of periods without agitation on PLT mitochondrial function, blood gases, and activation.
  
  
  METHODS
  Apheresis PLT units (n = 12) were collected using a cell separator and each was equally divided among five storage bags (50 mL of PLT suspension in 300-mL nominal volume containers). Four bags were held without agitation for 24, 48, 72, and 96 hours in a standard shipping box at room temperature and the fifth bag was continuously agitated. PLTs were assayed for standard in vitro PLT assays as well as for mitochondrial membrane potential (MMP), accumulation of reactive oxygen species, Annexin V binding, mitochondrial mass, and activity of mitochondrial reduction power (MRP) immediately after removal of units from the shipping container on Days 1, 2, 3, 4, and 7.
  
  
  RESULTS
  Increasing periods without agitation resulted in increased superoxide anion generation and PLT activation as well as reduced PLT MMP and MRP. Increasing periods without agitation resulted in increasing Annexin V binding. PLTs that had undergone periods without agitation showed increased oxygen and carbon dioxide levels immediately after storage without agitation. The superoxide anion generation was highly correlated with the loss of MMP, increasing Annexin V binding, and pH decline.
  
  
  CONCLUSIONS
  PLTs, if stored without agitation, produce a lesion that leads PLTs to apoptosis. The severity of the lesion depends on the length of the period without agitation. Prolonged periods without agitation induce formation of superoxides and depolarization of MMP along with a presentation of apoptotic markers.
authors.affiliation: American Red Cross Biomedical Services, Holland Laboratory, Blood Components Development, Rockville, Maryland 20855, USA. skripcha@usa.redcross.org
authors.forename: Andrey
authors.initials: A
authors.lastname: Skripchenko
authors.forename: Andrew
authors.initials: A
authors.lastname: Myrup
authors.forename: Dedeene
authors.initials: D
authors.lastname: Thompson-Montgomery
authors.forename: Helen
authors.initials: H
authors.lastname: Awatefe
authors.forename: Gary
authors.initials: G
authors.lastname: Moroff
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Wagner
mesh_terms: D017304:Annexin A5 / Q000097:blood; D017209:Apoptosis; D001786:Blood Glucose / Q000032:analysis; D001792:Blood Platelets / Q000737:chemistry / Q000648:ultrastructure*; D001793:Blood Preservation / Q000379:methods*; D002245:Carbon Dioxide / Q000097:blood; D006801:Humans; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D007773:Lactates / Q000097:blood; D008563:Membrane Lipids / Q000032:analysis; D053078:Membrane Potential, Mitochondrial; D008928:Mitochondria / Q000502:physiology*; D009038:Motion; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D010100:Oxygen / Q000097:blood; D010718:Phosphatidylserines / Q000032:analysis; D015539:Platelet Activation; D017382:Reactive Oxygen Species / Q000097:blood; D013481:Superoxides / Q000097:blood
chemical_list: D017304:Annexin A5; D001786:Blood Glucose; D007773:Lactates; D008563:Membrane Lipids; D010718:Phosphatidylserines; D017382:Reactive Oxygen Species; D013481:Superoxides; D002245:Carbon Dioxide; D010100:Oxygen