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Plasmodium chabaudi, a rodent malaria parasite with a synchronous asexual cycle in the blood, depending on the host’s circadian rhythm, was desynchronized by modifying its normal timing: blood taken from a donor mouse in the morning was kept 8 h at +4°C and inoculated in the evening into naive mice. When the infection had become asynchronous (from day 4 to day 7) mice were treated with a single dose of chloroquine. The efficacy of chloroquine was lower in mice with an asynchronous infection than in the control mice with a normally synchronous infection.
The identified mutations in the pfcrt, dhfr and dhps genes of Plasmodium falciparum show a very high correlation with resistance to chloroquine, pyrimethamine and sulfadoxine, the drugs that are still used as malaria chemoprophylaxis or treatment. We undertook a molecular screening of 82 Polish P. falciparum isolates, mainly imported from different countries of sub-Saharan Africa to assess their molecular drug-resistance profiles. Only 4 isolates showed no mutations in the three analyzed gene fragments. In the remaining isolates from one to six mutations in one or more examined genes were found. Different mutations in the pfcrt, dhfr and dhps genes were found in ca. 76%, 80% and 70% of P. falciparum isolates, respectively. About forty our patients used chloroquine or pyrimethamine + sulfadoxine as malaria chemoprophylaxis and/or antimalarial treatment, but without success. In all but 5 of the P. falciparum isolates obtained from these persons, mutations associated to resistance of the parasite to chloroquine and the antifolate drugs were found.
As a result of widespread resistance to chloroquine (CQ) and sulphadoxine-pyrimethamine (SP), artemisinin-based combination therapy (ACT) has been recommended as a first-line anti-malarial regimen in Côte d’Ivoire since 2005. A thorough understanding of the molecular bases of P. falciparum resistance to existing drugs is therefore needed. The aims of this study were to analyze the in vitro sensitivity of P. falciparum field isolates from Abobo to CQ, pyronaridine (PYR) and dihydroartemisinine (DHA), and to investigate the polymorphisms associated with drug resistance. The standard in vitro drug sensitivity microtechnique recommended by the WHO was used to assess the sensitivity of Plasmodium falciparum isolates collected in December 2006. The Pfcrt haplotype 76 was analysed by PCR-RFLP while Pfatpase 6 amplification products were sequenced. Associations between drug sensitivity and parasite gene polymorphisms were evaluated with Cohen’s kappa test. The correlation between the IC50 values for different drugs was assessed by the coefficient of determination (r2). Significance was assumed at p<0.05. Of 128 in vitro tests performed, 112 (87.5%) were successful. Of the isolates, 56.2% were resistant for CQ and 48% for PYR. One isolate (3.6%) demonstrated reduced DHA sensitivity (IC50 higher than 10 nM). The mutant K76T pfcrt codon, present in 90% of DNA fragments analyzed, was associated with CQ-R (ĸ=0.76). The N669Y (16.1%), D734Y (28.6%) and D734H (1.8%) isolates were found to have mutant Pfatpase6, however, these mutations were not associated with diminished DHA sensitivity (k=0.01). These high levels of antimalarial drug resistance in Abobo (Côte d’Ivoire) demand further studies of drug efficacy across the whole country.
Bistramide D and K were extracted from a New Caledonian marine colonial ascidian Lissoclinum bistratum and tested against Plasmodium berghei and P. vinckei petteri in mice. Bistramide D was able to reduce parasitaemia by almost 50% in P. berghei infected mice, but the ID₅₀ (>5 mg/kg) was very close to the LD₅₀ (8 mg/kg). In the same assay, bistramide K was less active than bistramide D. The mid-term trophozoite and the old trophozoite were shown to be the stages most sensitive to bistramide D in P. vinckei petteri infected mice. Therefore, bistramides are interesting experimental tools but do not present a major interest as potential antimalarial drugs.
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