Diagnosis of hemolytic disease by electrophoresis of erythrocyte lactate dehydrogenase isoenzymes on cellulose acetate or Agarose.

1981 ◽  
Vol 27 (8) ◽  
pp. 1453-1455 ◽  
Author(s):  
F Van Lente ◽  
A Marchand ◽  
R S Galen
Keyword(s):  
Lactate Dehydrogenase ◽  
Cellulose Acetate ◽  
Predictive Value ◽  
In Vitro Models ◽  
Hemolytic Disease ◽  
Cellulose Acetate Electrophoresis ◽  
Lactate Dehydrogenase Isoenzymes

Abstract We determined the LD-1/LD-2 isoenzyme ratio in hemolysates of erythrocytes by electrophoresis on cellulose acetate and on agarose. A ratio exceeding 1.0 was found with the former but not the latter. Results were similar for in vitro models of hemolytic disorders. Using cellulose acetate electrophoresis, we determined the predictive value of data on total LD activity and of the LD-1/LD-2 ratio in diagnosis of hemolytic disease in 100 patients. The sensitivity of the "flipped" LD-1/LD-2 ratio was only 58%, the specificity was 93%, and the predictive value was 74% for diagnosis of hemolytic disease. A normal total LD activity is highly predictive (92%) for ruling at the presence of hemolytic disease.

Effect of reticulocytosis on lactate dehydrogenase isoenzyme distribution in serum: in vivo and in vitro studies

1990 ◽  
Vol 36 (9) ◽  
pp. 1638-1641 ◽  
Author(s):  
S C Kazmierczak ◽  
W J Castellani ◽  
F Van Lente ◽  
E D Hodges ◽  
B Udis
Keyword(s):  
Flow Cytometry ◽  
Lactate Dehydrogenase ◽  
Cell Types ◽  
In Vitro Studies ◽  
Hemolytic Disease ◽  
Dehydrogenase Isoenzyme ◽  
Disease Analysis

Abstract We investigated the effect of reticulocytosis on the lactate dehydrogenase (LD; EC 1.1.1.27) isoenzyme LD1/LD2 ratio in patients with and without evidence of hemolytic disease. Analysis of sera from patients with reticulocytosis and in vivo hemolysis showed a mean LD1/LD2 ratio of 0.92 compared with a ratio of 0.69 in patients with in vivo hemolysis and normal reticulocyte counts. Determination of LD isoenzymes in erythrocyte lysate revealed significantly increased LD1/LD2 ratios for patients with marked reticulocytosis compared with those for patients with normal-to-minimal increases in reticulocytes. Finally, separation of mature erythrocytes and reticulocytes by flow cytometry revealed marked differences in the LD1/LD2 isoenzyme distribution between these two cell types. The ability of hemolysis to cause a "flipped" LD1/LD2 ratio is dependent on the proportion of the hemolyzed cells that are reticulocytes.

scholarly journals Adsorptive pinocytosis of 125I-labelled lactate dehydrogenase isoenzymes H4 and M4 by rat yolk sacs incubated in vitro

1981 ◽  
Vol 198 (3) ◽  
pp. 587-593 ◽  
Author(s):  
T Kooistra ◽  
K E Williams
Keyword(s):  
Lactate Dehydrogenase ◽  
Hydrophobic Interactions ◽  
Dissociation Constants ◽  
Yolk Sac ◽  
Sinusoidal Cells ◽  
Membrane Complex ◽  
Liver Sinusoidal Cells ◽  
Lactate Dehydrogenase Isoenzymes

The pinocytic uptake of 125I-labelled porcine lactate dehydrogenase isoenzymes H4 and M4 by 17.5-day rat visceral yolk sac incubated in vitro was saturable and binding obeyed Michaelis-Menten kinetics. The uptake characteristics of the two isoenzymes were very similar. For the H4 and the M4 isoenzymes, the dissociation constants of the protein-plasma-membrane complex were 0.62 microM and 0.84 microM respectively, and the maximum rates of uptake 0.13 and 0.26 nmol/mg of yolk-sac protein per h respectively. These findings contrast with those from studies in vivo, which show the M4 form is taken up by rat liver sinusoidal cells at a much higher rate than the H4 form, and point to different recognition systems for the adsorptive pinocytosis of simple non-conjugate proteins in yolk-sac epithelial cells and liver sinusoidal cells. Competition experiments indicate that binding of the H4 isoenzyme to the yolk-sac cells is restricted to hydrophobic interactions, whereas the binding of the M4 isoenzyme involves hydrophobic as well as positively charged sites on the protein molecules.

Falsely negative laboratory diagnosis for myocardial infarction owing to the concurrent presence of macro creatine kinase and macro lactate dehydrogenase.

1982 ◽  
Vol 28 (12) ◽  
pp. 2434-2437 ◽  
Author(s):  
M R Pudek ◽  
B E Jacobson
Keyword(s):  
Myocardial Infarction ◽  
Creatine Kinase ◽  
Lactate Dehydrogenase ◽  
Cellulose Acetate ◽  
Gel Filtration ◽  
Laboratory Diagnosis ◽  
Cellulose Acetate Electrophoresis

Abstract Macro creatine kinase (CK, EC 2.7.3.2) and macro lactate dehydrogenase (LD, EC 1.1.1.27) were both present in the serum of a 70-year-old woman with myocardial infarction. This interfered with the interpretation of the CK and LD isoenzyme analyses. Gel filtration and immunoprecipitation showed that the macro CK consisted of IgG and CK and the macro LD of IgG and LD. The IgG in this patient bound both MB and BB isoenzymes of CK, resulting in a macro CK complex that co-migrated with CK-MM on cellulose acetate electrophoresis. This situation led to a falsely negative laboratory diagnosis for myocardial infarction.

Evaluation of in-vivo animal and in-vitro models for prediction of dermal absorption in man

2008 ◽  
Vol 27 (4) ◽  
pp. 281-288 ◽  
Author(s):  
I Jakasa ◽  
S Kezic
Keyword(s):  
Animal Models ◽  
Human Skin ◽  
Predictive Value ◽  
Percutaneous Absorption ◽  
In Vitro Models ◽  
Alternative Methods ◽  
In Vitro Assays ◽  
Rat Skin

Risk assessment of dermal exposure to chemicals requires percutaneous absorption data to link the external exposure to the systemic uptake. The most reliable data on percutaneous absorption are obtained from in-vivo human volunteer studies. In addition to ethical constrains, the conduct of these studies is not feasible for the large number of industrial chemicals in use today. Therefore, there is an increasing need for alternative methods to determine percutaneous absorption such as in-vitro assays and methods performed in vivo in experimental animals. In this article, recent comparative in-vitro and in-vivo studies on percutaneous absorption have been addressed with emphasis on the factors that may affect the predictive value of the in-vitro models. Furthermore, the use of animal models, in particular the rat skin, in prediction of percutaneous absorption in the human skin has been reviewed. In-vitro assays showed to be largely influenced by the experimental circumstances, such as type and thickness of the skin, receptor fluid, and the way in which percutaneous absorption is calculated. Rat skin showed consistently to be more permeable than human skin. However, the difference between human and rat skin does not show a consistent pattern between chemicals hampering prediction of human percutaneous absorption. To increase predictive value of in-vitro and animal models, the influence of experimental factors on the percutaneous absorption should be systematically investigated by comparison with human in-vivo data, resulting in more prescriptive guidelines.

Analysis and applicability of different in vitro models of Glioblastoma multiforme

2014 ◽  
Vol 226 (06) ◽  
Author(s):  
D William ◽  
M Linnebacher ◽  
CF Classen
Keyword(s):  
Glioblastoma Multiforme ◽  
In Vitro Models
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