Among the quassinoids isolated from Eurycoma longifolia Jack, eurycomanone was identified as the most potent and toxic inhibitor of the chloroquine-resistant Gombak A isolate of Plasmodium falciparum. Several diacylated derivatives of eurycomanone, 1,15-di-O-isovaleryleurycomanone, 1,15-di-O-(3,3-dimethylacryloyl)- eurycomanone and 1,15-di-O-benzoyleurycomanone were synthesized by direct acylation with the respective acid chlorides. The monoacylated 15-O-isovaleryleurycomanone was synthesized by selective protection of the other hydroxy groups of eurycomanone with trimethylsilyl trifluoromethanesulphonate to enable the exclusive acylation of its C-15 hydroxy group. This was followed by the removal of the protecting groups with citric acid. The diacylated eurycomanones exhibited lower antiplasmodial activity against the Gombak A isolates and lower toxicity in the brine shrimp assay when compared to eurycomanone. In contrast, the monoacylated derivative displayed comparable antiplasmodial potency to eurycomanone, but its toxicity was reduced. Thus, preliminary studies of the synthesized acylated eurycomanones have shown that acylation only at the C-15 hydroxy group may be worthy of further antimalarial investigation.
Anopheles mosquitos were surveyed using three trapping technics in four longhouse settlements and their respectively farming zone in western Sarawak, Malaysia. The study area was mountainous with tropical rain forest. An. leucosphyrus and An. donaldi were predominant in the farm huts. An. tessellatus and An. subpictus were more abundant in the village settlements. In both ecotypes, human baited traps yielded a significantly greater proportion of Anopheles mosquito than CDC light traps and landing biting catches. Circumsporozoite antigen positively rate, mosquito survival rate and parasite rate showed that malaria transmission is more intense in farm huts than in longhouse settlements. The entomological inoculation rate of An. donaldi and An. leucosphyrus in farm huts was 0.035 and 0.023, respectively. No sporozoite infections were observed in the main settlements.
The gene encoding the 195,000-Da major merozoite surface antigen (gp195) of the FUP (Uganda-Palo Alto) isolate of Plasmodium falciparum, a strain widely used for monkey vaccination experiments, has been cloned and sequenced. The translated amino acid sequence of the FUP gp195 protein is closely related to the sequences of corresponding proteins of the CAMP (Malaysia) and MAD-20 (Papua New Guinea) isolates and more distantly related to those of the Wellcome (West Africa) and K1 (Thailand) isolates, supporting the proposed allelic dimorphism of gp195 within the parasite population. The prevalence of dimorphic sequences within the gp195 protein suggests that many gp195 epitopes would be group-specific. Despite the extensive differences in amino acid sequence between gp195 proteins of these two groups, the hydropathy profiles of proteins representative of both groups are very similar. The conservation of overall secondary structure shown by the hydropathy profile comparison indicates that gp195 proteins of the various P. falciparum isolates are functionally equivalent. This information on the primary structure of the FUP gp195 protein will enable us to evaluate the possible roles of conserved, group-specific and variable epitopes in immunity to the blood stage of the malaria parasite.
Malaria, a devastating disease, has claimed numerous lives and caused considerable suffering, with young children and pregnant women being the most severely affected group. However, the emergence of multidrug-resistant strains of Plasmodium and the adverse side effects associated with existing antimalarial drugs underscore the urgent need for the development of novel, well-tolerated, and more efficient drugs to combat this global health threat. To address these challenges, six new hydantoins derivatives were synthesized and evaluated for their in vitro antiplasmodial activity. Notably, compound 2c exhibited excellent inhibitory activity against the tested Pf3D7 strain, with an IC50 value of 3.97 ± 0.01 nM, three-fold better than chloroquine. Following closely, compound 3b demonstrated an IC50 value of 27.52 ± 3.37 µM against the Pf3D7 strain in vitro. Additionally, all the hydantoins derivatives tested showed inactive against human MCR-5 cells, with an IC50 value exceeding 100 μM. In summary, the hydantoin derivative 2c emerges as a promising candidate for further exploration as an antiplasmodial compound.
Background: Many proteins released by cells to the blood and other fluids are glycoproteins. One set of glycoproteins carry the ABO blood group determinants and glycoproteins have been shown to be vital in determining the structure and organization of plasma membranes. There is evidence suggesting their important role in cell-to-cell contact, adhesion, hormone interaction and vital transformation. Differences in proteins and glycoproteins in the different human blood groups may influence the invasion process of Plasmodium falciparum. The objectives of the study were to determine whether there are any changes in proteins and glycoproteins of red blood cells upon infection by P. falciparum and whether these protein and glycoprotein changes differ in the various ABO blood groups.
Methods: A Malaysian strain of P. falciparum was cultured in vitro in red blood cells from A, B, O and AB blood groups. Protein and glycoprotein profiles of uninfected and P. falciparum- infected red blood cells from the different human ABO blood groups were analyzed by SDS-PAGE. For protein bands, the gels were stained with Coomassie blue while glycoproteins were visualized following staining of gels using GelCode ® Glycoprotein Staining Kit.
Results: Cell membranes of P. falciparum infected erythrocytes from different ABO blood groups have different glycoprotein profiles compared to uninfected cells. All the infected samples showed a prominent protein band of molecular weight 99 kDa which was not present in any of the uninfected samples while a 48 kDa band was seen in four out of the seven infected samples. The erythrocyte cell membranes of A and AB blood groups showed different glycoprotein profiles upon infection with P. falciparum when compared to those from blood groups B and O.
Conclusion: The two glycoproteins of molecular weights 99 kDa and 48 kDa should be further studied to determine their roles in the pathogenesis of malaria and as potential targets for drug and vaccine development.
There are seven known species of Plasmodium spp. that can infect humans. The human host can mount a complex network of immunological responses to fight infection and one of these immune functions is phagocytosis. Effective and timely phagocytosis of parasites, accompanied by the activation of a regulated inflammatory response, is beneficial for parasite clearance. Functional studies have identified specific opsonins, particularly antibodies and distinct phagocyte sub-populations that are associated with clinical protection against malaria. In addition, cellular and molecular studies have enhanced the understanding of the immunological pathways and outcomes following phagocytosis of malaria parasites. In this review, an integrated view of the factors that can affect phagocytosis of infected erythrocytes and parasite components, the immunological consequences and their association with clinical protection against Plasmodium spp. infection is provided. Several red blood cell disorders and co-infections, and drugs that can influence phagocytic capability during malaria are also discussed. It is hoped that an enhanced understanding of this immunological process can benefit the design of new therapeutics and vaccines to combat this infectious disease.
According to the report of the Intergovernmental Panel on Climate Change (IPCC), Malaysia will experience an increase of 3-5°C in the future. As the development of the malaria parasite, Plasmodium falciparum, is sensitive to temperature, we investigated, using computer models, the effect of increase of 3º and 5ºC on the possible changes in the epidemiology of malaria transmission of P. falciparum in Malaysia. Four environmentally different locations were selected: Kuala Lumpur (KL), Cameron Highlands (CH), Kota Kinabalu (KK) and Kinabalu Park (KP). The extrinsic incubation period (EIP) was estimated using hourly temperatures and the mean daily temperatures. The EIP values estimated using the mean daily temperature were lower than those computed from hourly temperatures in warmer areas (KL, KK), but higher in the cooler areas (CH, KP). The computer simulations also indicated that the EIP will be decreased if the temperature was raised by 3º or 5ºC, with the effect more pronounced for the greater temperature increase, and for the cooler places. The vector cohort that is still alive at a time to transmit malaria (s(EIP)) also increased when the temperature was raised, with the increase more pronounced in the cooler areas. This study indicates an increase in temperature will have more significant effect in shortening the EIP in a cooler place (eg CH, KP), resulting in a greater s(EIP), and consequently increasing the transmission intensity and malaria risk. A temperature increase arising from the global climate change will likely affect the epidemiology of malaria in Malaysia, especially in the cooler areas.
To assess the public health importance of malaria on Banggi Island, Sabah, baseline epidemiological and entomological data were obtained in a study of three villages. These data were used to model the transmission of malaria using a non-seasonal version of the deterministic model of Dietz, Molineaux and Thomas. The model provided a satisfactory description of prevalence rates of Plasmodium falciparum parasitaemia. Modifications to the basic model enable the effects of mass chemotherapy with various combinations of schizonticidal and gametocidal drugs to be simulated. In this way, the relative merits of different procedures of mass drug administration can be compared. The fitted model is also used to examine the relationship between the overall prevalence of infection and the vectorial capacity, and to predict the consequences of a reduction in the size of the vector population.
Comparing patterns of genetic variation at multiple loci in the genome of a species can potentially identify loci which are under selection. The large number of polymorphic microsatellites in the malaria parasite Plasmodium falciparum are available markers to screen for selectively important loci. The Pfs48/45 gene on Chromosome 13 encodes an antigenic protein located on the surface of parasite gametes, which is a candidate for a transmission blocking vaccine. Here, genotypic data from 255 P. falciparum isolates are presented, which show that alleles and haplotypes of five single nucleotide polymorphisms (SNPs) in the Pfs48/45 gene are exceptionally skewed in frequency among different P. falciparum populations, compared with alleles at 11 microsatellite loci sampled widely from the parasite genome. Fixation indices measuring inter-population variance in allele frequencies (F(ST)) were in the order of four to seven times higher for Pfs48/45 than for the microsatellites, whether considered (i) among populations within Africa, or (ii) among different continents. Differing mutational processes at microsatellite and SNP loci could generally affect the population structure at these different types of loci, to an unknown extent which deserves further investigation. The highly contrasting population structure may also suggest divergent selection on the amino acid sequence of Pfs48/45 in different populations, which plausibly indicates a role for the protein in determining gamete recognition and compatibility.
Matched MeSH terms: Plasmodium falciparum/genetics*; Plasmodium falciparum/growth & development
Dihydropteroate synthase (dhps) and dihydrofolate reductase (dhfr) alleles were typed in 67 Malaysian Plasmodium falciparum isolates. The isolates were collected from two geographically distinct locations: 51 from Sabah, Malaysian Borneo, where sulfadoxine/pyrimethamine (SDX/PYR) is used to treat uncomplicated malaria and 16 from Peninsular Malaysia where in vivo resistance to SDX/PYR has been reported. A total of seven dhps alleles were identified with no significant difference in allele frequency between the 2 populations. Two of the dhps alleles described here have not been previously reported. Four dhfr alleles were detected in 67 P. falciparum isolates. Eighty-seven percent of the isolates from the Peninsula, where clinical SDX/PYR failure has been reported, had dhfr alleles with triple point mutations while all of the isolates from Sabah had dhfr alleles with 2 or less point mutations. The difference in dhfr allele frequency between the two populations was highly significant. There was no correlation between in vitro PYR response and accumulation of dhfr point mutations.
Thirty-two patients reporting to the Lundu District Hospital, Sarawak, Malaysian Borneo, with uncomplicated falciparum malaria were recruited into a multifaceted study to assess treatment response. Following combined chloroquine and sulphadoxine/pyrimethamine treatment the patients were followed for 28 days according to the World Health Organisation in vivo drug response protocol. The in vivo study revealed that 13 (41%) of the patients had a sensitive response to treatment, five (16%) cleared asexual stage parasites but had persistent gametocytes, 11 (34%) had RI type resistance and three (9%) had RII type resistance requiring quinine intervention before day 7 for parasite clearance. Although clinically insignificant, patients with persistent gametocytes, surviving chloroquine and sulphadoxine/pyrimethamine treatment during maturation, were placed in the reduced response to treatment group for analysis. Allelic typing detected 100% prevalence of the pfcrt K76T marker associated with chloroquine resistance and 78% prevalence of the pfdhfr NRNL haplotype associated with sulphadoxine/pyrimethamine treatment failure. High serum chloroquine levels and pfdhfr haplotypes with
Malaria remains a disease of underdeveloped and remote regions of the world. The application of polymerase chain reaction (PCR) technology to malaria epidemiology has the potential for increasing our knowledge and understanding of this disease. In order to study malaria in all geographical locations it is important that specimen collection and DNA extraction for PCR be kept simple. Here we report a method for extracting DNA from dried blood spots on filter paper which is capable of detecting one Plasmodium falciparum and two Plasmodium vivax parasites/microliter of whole blood by nested PCR without compromising the simplicity of specimen collection or DNA extraction.