Document Detail

Reversible cross-linking and CO treatment as an approach in red cell stabilization.
MedLine Citation:
PMID:  10924266     Owner:  NLM     Status:  MEDLINE    
We explored the use of the reversible cross-linking reagent dimethyl 3,3-dithiobispropionimidate (DTBP) in combination with CO treatment as an approach to stabilizing erythrocyte structure and function. Erythrocytes were cross-linked with different concentrations of DTBP for different times. DTBP increased erythrocyte osmotic stability, blocked lysolecithin-induced echinocytosis, and decreased erythrocyte deformability in a concentration- and time-dependent manner. Reversal of the cross-linking with the reducing agent dithioerythritol (DTE) restored osmotic fragility and response to lysolecithin as well as deformability. Complete reversal, however, is a function of the DTBP concentration and the time of cross-linking. The effects of cross-linking with 5 mM DTBP for 1 h were completely reversible after treatment with 10 mM DTE for 20 min. Longer incubation times or higher concentrations of DTBP resulted in partial reversal by DTE of the effects produced by DTBP. Cross-linking and reversal only slightly reduced the ATP content. The hemoglobin contained in the cross-linked and reversed cells could still undergo reversible oxygenation and deoxygenation. Erythrocytes were pretreated with CO, cross-linked with 5 mM DTBP for 1 or 3 h, loaded with a solution containing 500 mM glucose for 24 h, and freeze-dried in a medium containing 15% (w/v) albumin. Rehydration followed in distilled water. Complete recovery, measured as the percentage of free hemoglobin, was achieved for cells cross-linked with 5 mM DTBP for 3 h and freeze-dried to a final water content of 10-15%. Non-cross-linked cells lysed 100% on rehydration in distilled water. No methemoglobin (MetHb) formation as a result of freeze-drying was detected in CO-treated cells. In non-CO-treated cells 20% of the Hb was converted to MetHb.
I Bakaltcheva; S Leslie; V MacDonald; B Spargo; A Rudolph
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Publication Detail:
Type:  In Vitro; Journal Article; Research Support, U.S. Gov't, Non-P.H.S.    
Journal Detail:
Title:  Cryobiology     Volume:  40     ISSN:  0011-2240     ISO Abbreviation:  Cryobiology     Publication Date:  2000 Jun 
Date Detail:
Created Date:  2000-10-06     Completed Date:  2000-10-06     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0006252     Medline TA:  Cryobiology     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  343-59     Citation Subset:  IM    
Copyright Information:
Copyright 2000 Academic Press.
Geo-Centers, Inc., 1801 Rockville Pike, Suite 405, Rockville, Maryland 20852, USA.
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MeSH Terms
Adenosine Triphosphate / blood
Blood Preservation / methods*
Carbon Monoxide / pharmacology
Cross-Linking Reagents / pharmacology
Cryopreservation / methods*
Dithioerythritol / pharmacology
Erythrocyte Deformability / drug effects
Erythrocytes* / cytology,  drug effects,  metabolism
Freeze Drying
Imidoesters / pharmacology
Osmotic Fragility / drug effects
Reg. No./Substance:
0/Cross-Linking Reagents; 0/Imidoesters; 56-65-5/Adenosine Triphosphate; 59012-54-3/dimethyl dithiobispropionimidate; 630-08-0/Carbon Monoxide; 6892-68-8/Dithioerythritol

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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