Antiparasitic drugs pdf
ZnCl2 0. The MT content was determined by the radioactive cadmium method as described in the Materials and Methods section.
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Effect of nifurtimox, benznidazole and zinc chloride upon metallothionein content in Trypanosoma cruzi Table II shows the effect of nifurtimox and benznidazole upon MT concentration in the Tulahuén epimastigotes. MTs cannot bind cadmium or other metals when their thiol groups are oxidized or conjugated with reactive electrophiles. Figure 2. Chromatographic analysis of Metallothioneins of Férgek elleni eszköz cruzi.
The black continuous line represents the elution pattern of T. The black discontinuous line represents monobromobimane-derivatized T. The gray discontinuous line represents monobromobimane-derivatized horse kidney standard MT. The T. See Materials and Methods section for details.
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Chromatographic analysis of metallothioneins of Trypanosoma cruzi Figure 2 shows the elution pattern of T. Monobromobimane derivatized MT from T. Similarly, the native MT detected with the Cd method also showed a bimodal pattern.
Standard monobromobimane-derivatized MT from horse kidney eluted with one peak at fractions. Both peaks of native MT showed no measurable absorbance at nm and negligible reaction in the Lowry protein determination indicating a low concentration or absence of aromatic amino acids.
When T. The addition of mercaptoethanol recovered the original value for MTs, due to the reduction of disulfide bonds formed. This standard demonstrates that T. The proteolytic degradation of MT during the homogenization process antiparasitic drugs pdf inhibited by the addition of antiproteases, excluding this possibility as an explanation for the two peaks observed in Figure 2 when determined by antiparasitic drugs pdf radioactive cadmium method or by monobromobimane derivatization.
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Figure 3. Effect of nifurtimox and benznidazole upon MT-like proteins of Trypanosoma cruzi epimastigotes. Line 1 is the standard, underivatized horse kidney metallothionein. Line 2 is the same standard but monobromobimane-derivatized.
Lines 3 and 4 correspond to 1. Line 7 is the T. Further details in the Materials and Methods section.
Both panels, A and B, correspond to the same gel. Line 1 is the underivatized, standard horse kidney metallothionein. There are no apparent differences between control and treatment lines in the Coomasie blue gel. Figure 3Blines 2B and 7B, show that T. Lines 3B to 6B show that treatment of the parasites with both nifurtimox and benznidazole produces disappearance of this fluorescent band corresponding to MT.
When the two peaks of T. This indicates that both peaks have a very similar molecular weight.
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Nevertheless, very low-intensity, small fluorescence bands in the high molecular weight region persist throughout the treatment and control lines. In the Cl-Brenner clone the MT content did not change significantly between the exponential and the stationary phase.
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At present we have no explanation for this observation. However, we might speculate that in the CL-Brenner clone the MT protein regulatory processes synthesis and degradation do not differ in the two growth phases.
Nevertheless, other explanations are possible.
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T SH 2 is the major thiol-containing compound in T. The reaction of the nifurtimox or benznidazole's electrophilic metabolites with the MT's thiol groups most probably causes this effect.
Such a reaction is similar to the conjugation of the nifurtimox or benznidazole's electrophilic metabolites with the thiol groups of T SH 2 and GSH Repetto et al.
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MTs are inducible by several antiparasitic drugs pdf including oxidative and physical stress. The radioactive cadmium method only detects free reduced MTs and consequently, does not detect MTs conjugated with drug metabolites Table II. Also, MT conjugated with drug metabolites does not react with monobromobimane and thus does not produce fluorescence Fig.
MTs are better radical scavengers than GSH. The rate constant for the reaction of MT with hydroxyl radicals and electrophilic agents is about three hundred times higher than with GSH Sato and Bremner, Nitroheterocyclic agents, such as nifurtimox, produce hydroxyl radicals Docampo and Moreno,which may hpv ano uomo with thiol groups.
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In antiparasitic drugs pdf respect, we may see in Figure 3 that the parasites' treatment with both nifurtimox and benznidazole produces disappearance of the fluorescence bands indicating reaction between MT and drug metabolites.
Antiparasitic drugs pdf, Figure 3 shows that low-intensity fluorescence bands in the high molecular weight range of the gel do not disappear, indicating the presence of proteins with less reactive thiol groups. In previous reports we showed evidence that correlates T SH 2 and GSH concentration with the susceptibility to nifurtimox and benznidazole of several T. In this report, we found no direct evidence for the same role of MTs in T. Nevertheless, in other organisms, MTs have been shown to have this property Perego et al.
It is probable that MTs and other thiol-containing compounds, such as ovothiols Steenkamp and Spies, ; Ariyanayagam and Fairlamb,may contribute to protection of the parasite against oxidative stress. We conclude that MT-like proteins are present in T. In addition, these proteins may contribute to the resistance of drugs that act producing free radicals or electrophilic metabolites.
As stated in the introduction section of this paper, MTs have been isolated from other protozoa. However, this is the first time antiparasitic drugs pdf MTs are identified in parasites of medical importance. The authors gratefully acknowledge the critical review of this paper by Prof.
Res R Effect and mode of action of nitroimidazole and nitrofuran derivatives.
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