The
diaphorase activity of PfFNR in the presence of the Stemona alkaloids at 100 ppm is shown in Table 2.
Thus NADH
diaphorase helps in curbing this effect as has been demonstrated by this work.
Parent to child transmission and longstanding cyanosis suggests the possibility of haemoglobin M, whereas congenital cyanosis in siblings is more consistent with the deficiency of NADH-cytochrome b5 reductase (
diaphorase) (9).
Alcohol dehydrogenase, NAD, ADP-ribosyl cyclase, nicotinamide, resazurin,
diaphorase, bovine serum albumin (BSA), 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT), bicine, riboflavin 5'-mono-phosphate (FMN), phenazine ethosulfate (PES), ethylenediaminetetraacetic acid (EDTA)-Na, alcohol dehydrogenase (ADH), and 8-Br-cADPR were purchased from Sigma-Aldrich (St.
Briefly, the yellow MTT tetrazolium converts to purple formazan upon removal of the bromide by
diaphorase flavoprotein enzyme present in metabolically active human sperm (37--39).
Forster, "Histochemial staining and qualification of dihydrolipoamide dehydrogenase
diaphorase activity using blue native PAGE," Electrophoresis, vol.
(58) [NAD.sup.+]-reducing hydrogenase from hydrogen bacterium, Ralstonia eutropha H16 has four more subunits, two of which constitute
diaphorase to mediate electron transfer between the core hetero dimer and [NAD.sup.+]/NADH.
The final assay mixture was comprised of 110 mM lactate, 3.35 mM [NAD.sup.+], 350 [micro]M resazurin and 2.2 units/mL of
diaphorase in 3 mM Tris/30 mM HEPES/10 mM NaCl buffer (pH 7.4), and 0.45% Triton X100 for total LDH.
In addition, hereditary methemoglobinemias due to NADH dependent cytochrome reductase (NADH dependent methemoglobine reductase = NADH dependent
diaphorase) deficiency, which was assayed only by us so far in Turkey with probable dominant inheritance and Hemoglobin M cases were reported on several journals (5-11).
2DG, hexoki-nase, G6PDH (glucose-6-phosphate dehydrogenase),
diaphorase, rezazurine, ATP, [NADP.sup.+], and all the primers were obtained from Sigma-Aldrich, Bangalore, India.
These decreases were followed by increases in levels of phase II components such as glutathione (GSH), glutathione-S-transferase (GST), and NAD(P)H:quinone oxidoreductase (DT
diaphorase).