scholarly journals THE EFFECT OF MENADIONE AND PHENAZINE METHOSULFATE ON THE TETRAZOLIUM REDUCTION SYSTEM UNDER HISTOCHEMICAL CONDITIONS

1964 ◽  
Vol 12 (11) ◽  
pp. 797-804 ◽  
Author(s):  
T. HASHIMOTO ◽  
J. S. KALUZA ◽  
M. S. BURSTONE
Keyword(s):  
Nitro Blue Tetrazolium ◽  
Soluble Factor ◽  
Phenazine Methosulfate ◽  
Frozen Sections ◽  
Hydrogen Acceptor ◽  
Variable Effect ◽  
Isocitric Dehydrogenase ◽  
Blue Tetrazolium ◽  
Fresh Frozen ◽  
Tetrazolium Reduction

The influence of phenazine methosulfate and menadione on the activity of DPNH- and TPNH-tetrazolium reductase, succinic, lactic, and triphosphopyridine nucleotide-linked isocitric dehydrogenase were studied with reference to applied histochemistry. Fresh frozen sections of various tissues were employed in conjunction with nitro blue tetrazolium as the hydrogen acceptor. Phenazine methosulfate was found to produce a variable effect upon staining, enhancing it or suppressing it according to incubation conditions. Suppression was most pronounced with higher concentrations. Menadione had no suppressive influence and was useful in increasing tetrazolium reduction. Its usefulness is, however, limited unless a soluble factor, possibly DT-(DPNH-TPNH)-diaphorase, can be prevented from diffusing into the medium, and "nothing dehydrogenase" is selectively inactivated.

Heterogeneity in superoxide production as measured by nitro blue tetrazolium reduction from individual PAM

1991 ◽  
Vol 260 (6) ◽  
pp. L464-L470 ◽  
Author(s):  
K. A. DiGregorio ◽  
E. V. Cilento ◽  
R. C. Lantz
Keyword(s):  
Cell Volume ◽  
Initial Rate ◽  
Single Cells ◽  
Nitro Blue Tetrazolium ◽  
Blue Tetrazolium ◽  
Cell Data ◽  
O2 Production ◽  
Cell Volumes ◽  
Tetrazolium Reduction ◽  
Small Subpopulation

Heterogeneity in superoxide (O2-.) production (as determined by the reduction of nitro blue tetrazolium to a diformazan precipitate), cell volume, and cell spreading were measured from single rat pulmonary alveolar macrophages (PAM). Of the 330 cells that produced O2-. due to stimulation by adherence, the maximum diformazan produced (MAX) varied between 1.8 and 47.2 fmol and the maximum (initial) rate of diformazan production (R) varied between 2.2 and 81.7 X 10(-3) fmol/s. Only six PAM of 336 analyzed failed to produce O2-. suggesting that O2-. production is not a specific function of a small subpopulation of PAM but rather that all PAM are capable of producing O2-.. Importantly, O2-. production by single cells showed extensive heterogeneity that did not correlate (r2 less than 0.12) with individual cell volumes, suggesting that the observed heterogeneity may not be due to differences in maturation and/or differentiation. However, when the single cell data were grouped into cell volume subfractions small but increasing linear trends (r2 greater than 0.81) in MAX and R were found. This discrepancy suggests that PAM heterogeneity in O2-. fraction cannot be estimated adequately by measurements on subpopulations of cells.

scholarly journals Interactions of some acceptors with superoxide anion radicals formed by the NADPH-specific flavoprotein in rat liver microsomal fractions

1976 ◽  
Vol 154 (2) ◽  
pp. 307-310 ◽  
Author(s):  
V Mishin ◽  
A Pokrovsky ◽  
V V Lyakhovich
Keyword(s):  
Rat Liver ◽  
Superoxide Anion ◽  
Microsomal Fraction ◽  
Nitro Blue Tetrazolium ◽  
Blue Tetrazolium ◽  
Liver Microsomal Fraction ◽  
Dependent Pathway ◽  
Tetrazolium Reduction ◽  
Superoxide Anion Radicals ◽  
Anion Radicals

In rat liver microsomal fractions oxidation of adrenaline was effected by superoxide anion radicals (O2-), whereas cytochrome c, 2,6-dichlorophenol-indophenol and ferricyanide accepted electrons from NADPH-specific flavoprotein only directly. Nitro Blue Tetrazolium was reduced both by O2- and by the direct acceptance of electrons. Elevation of pH and addition of menadione shift the Nitro Blue Tetrazolium reduction towards the O2--dependent pathway. From the values of the kinetic constants for interaction of adrenaline and Nitro Blue Tetrazolium with NADPH-specific flavoprotein, the rates of generation of O2- in rat liver microsomal fraction were determined.

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