Optimized Spectrophotometric Determination of Aldehyde Dehydrogenase Activity in Erythrocytes

1992 ◽  
Vol 38 (4) ◽  
pp. 584-588 ◽  
Author(s):  
R D Johnson ◽  
J Bahnisch ◽  
B Stewart ◽  
D J Shearman ◽  
J B Edwards
Keyword(s):  
Alcohol Consumption ◽  
Dehydrogenase Activity ◽  
Aldehyde Dehydrogenase ◽  
Biological Marker ◽  
Aldh Activity ◽  
Lactate Dehydrogenase Activity ◽  
Analytical Imprecision ◽  
Excess Alcohol Consumption ◽  
Aldehyde Dehydrogenase Activity

Abstract We describe a reliable and sensitive semiautomated spectrophotometric assay of aldehyde dehydrogenase (ALDH; EC 1.2.1.3) activity in erythrocytes. The hemolysate can be stabilized with sucrose, and the technique involves only microliters of hemolysate on a centrifugal analyzer. The use of microcolumns to remove interfering hemoglobin is avoided, and reproducibility of the assay has been improved by manipulating the inherent lactate dehydrogenase activity of erythrocytes by adding lactate and oxalate to the reaction mixture. These modifications have decreased the analytical imprecision of the assay, allowing a better appraisal of aldehyde dehydrogenase activity in erythrocytes as a biological marker of excess alcohol consumption. Erythrocytic ALDH activity was significantly less in 40 alcoholics than in 145 teetotallers (median activity 128 vs 219 mU/g of hemoglobin, respectively; P = 0.0001), indicating the potential of this assay as a useful marker of excess alcohol consumption.

Use of an “acetaldehyde clamp” in the determination of low-KM aldehyde dehydrogenase activity in H4-II-E-C3 rat hepatoma cells

Alcohol ◽  
2003 ◽  
Vol 31 (1-2) ◽  
pp. 19-24 ◽  
Author(s):  
Claudio Moncada ◽  
Nelson Fuentes ◽  
Alvaro Lladser ◽  
Gonzalo Encina ◽  
Amalia Sapag ◽  
...  
Keyword(s):  
Dehydrogenase Activity ◽  
Aldehyde Dehydrogenase ◽  
Hepatoma Cells ◽  
Rat Hepatoma ◽  
Rat Hepatoma Cells ◽  
Aldehyde Dehydrogenase Activity

scholarly journals Selection based on CD133 and high aldehyde dehydrogenase activity isolates long-term reconstituting human hematopoietic stem cells

Blood ◽  
2006 ◽  
Vol 107 (5) ◽  
pp. 2162-2169 ◽  
Author(s):  
David A. Hess ◽  
Louisa Wirthlin ◽  
Timothy P. Craft ◽  
Phillip E. Herrbrich ◽  
Sarah A. Hohm ◽  
...  
Keyword(s):  
Stem Cells ◽  
Hematopoietic Stem Cells ◽  
Dehydrogenase Activity ◽  
Aldehyde Dehydrogenase ◽  
Cell Selection ◽  
Aldh Activity ◽  
Hematopoietic Stem ◽  
Selection For ◽  
Aldehyde Dehydrogenase Activity

The development of novel cell-based therapies requires understanding of distinct human hematopoietic stem and progenitor cell populations. We recently isolated reconstituting hematopoietic stem cells (HSCs) by lineage depletion and purification based on high aldehyde dehydrogenase activity (ALDHhiLin- cells). Here, we further dissected the ALDHhi-Lin- population by selection for CD133, a surface molecule expressed on progenitors from hematopoietic, endothelial, and neural lineages. ALDHhiCD133+Lin- cells were primarily CD34+, but also included CD34-CD38-CD133+ cells, a phenotype previously associated with repopulating function. Both ALDHhiCD133-Lin- and ALDHhiCD133+Lin- cells demonstrated distinct clonogenic progenitor function in vitro, whereas only the ALDHhiCD133+Lin- population seeded the murine bone marrow 48 hours after transplantation. Significant human cell repopulation was observed only in NOD/SCID and NOD/SCID β2M-null mice that received transplants of ALDHhiCD133+Lin- cells. Limiting dilution analysis demonstrated a 10-fold increase in the frequency of NOD/SCID repopulating cells compared with CD133+Lin- cells, suggesting that high ALDH activity further purified cells with repopulating function. Transplanted ALDHhiCD133+Lin- cells also maintained primitive hematopoietic phenotypes (CD34+CD38-) and demonstrated enhanced repopulating function in recipients of serial, secondary transplants. Cell selection based on ALDH activity and CD133 expression provides a novel purification of HSCs with long-term repopulating function and may be considered an alternative to CD34 cell selection for stem cell therapies.

Diminished alcohol preference in transgenic mice lacking aldehyde dehydrogenase activity

2002 ◽  
Vol 12 (8) ◽  
pp. 621-626 ◽  
Author(s):  
Toyohi Isse ◽  
Tsunehiro Oyama ◽  
Kyoko Kitagawa ◽  
Koji Matsuno ◽  
Akiko Matsumoto ◽  
...  
Keyword(s):  
Transgenic Mice ◽  
Dehydrogenase Activity ◽  
Aldehyde Dehydrogenase ◽  
Alcohol Preference ◽  
Aldehyde Dehydrogenase Activity
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