TABLE 1.

Relation between TcAKR activity and Bz susceptibility in epimastigotes of CL Brener and Nicaragua strains of T. cruzi

Parameter	Value for the parameter by strain
	CL Brener (DTU VI)	Nicaragua (DTU I)
TcAKR expression (n-fold)a,i	3.45 ± 0.46	1.86 ± 0.51
NADPH-dependent enzymatic activity (nmol NADPH/min/mg)
AKRb,i	365.37 ± 25.64	137.31 ± 8.72
QORc	140.92 ± 13.84	44.58 ± 7.53
Bz reductased,i	16.01 ± 0.65	2.80 ± 0.73
NADH-dependent enzymatic activity (nmol NADH/min/mg)
Bz reductasee,i	19.43 ± 1.44	8.24 ± 1.31
Bz trypanocidal activity
IC50 (μM)f,i	10 ± 0.095	20.50 ± 0.30
ROS (RFU)g	4.58 ± 0.88	3.63 ± 0.22
ψmh	0.28 ± 0.02	0.22 ± 0.02

^aRatio of the densitometric values of TcAKR and TcCyp19 bands obtained in Western blot assays as shown in Fig. 2B.

^bAKR activity using 4-NBA as a substrate.

^cQOR activity using 9,10-PQ as a substrate.

^dBz reduction using NADPH as a cofactor.

^eBz reduction using NADH as a cofactor.

^fIC₅₀s calculated by linear regression analysis of the plot of the growth constant versus drug concentration (Fig. 2A).

^gDetection of intracellular ROS production by flow cytometry using the H₂DCF-DA probe of Bz-treated epimastigotes. Values correspond to the relative fluorescence units (RFU) calculated as the median fluorescence intensity (MFI) of Bz-treated parasites/MFI of untreated parasites.

^hMeasurement of change in mitochondrial membrane potential (ΔΨm) by flow cytometry using rhodamine 123 of Bz-treated epimastigotes. Values correspond to the relative fluorescence units (RFU) calculated as the median fluorescence intensity (MFI) of Bz-treated parasites/MFI of untreated parasites.

ⁱP < 0.05%, for the difference between results with CL Brener and Nicaragua.