scholarly journals Imported Malaria in Portugal: Prevalence of Polymorphisms in the Anti-Malarial Drug Resistance Genes pfmdr1 and pfk13

2021 ◽  
Vol 9 (10) ◽  
pp. 2045
Author(s):  
Debora Serrano ◽  
Ana Santos-Reis ◽  
Clemente Silva ◽  
Ana Dias ◽  
Brigite Dias ◽  
...  
Keyword(s):  
Public Health ◽  
Drug Resistance ◽  
Plasmodium Falciparum ◽  
Resistance Genes ◽  
Malaria Case ◽  
African Country ◽  
Imported Malaria ◽  
Wild Type ◽  
Close Relationship

Malaria is one of the ‘big three’ killer infectious diseases, alongside tuberculosis and HIV. In non-endemic areas, malaria may occur in travelers who have recently been to or visited endemic regions. The number of imported malaria cases in Portugal has increased in recent years, mostly due to the close relationship with the community of Portuguese language countries. Samples were collected from malaria-infected patients attending Centro Hospitalar Lisboa Ocidental (CHLO) or the outpatient clinic of Instituto de Higiene e Medicina Tropical (IHMT-NOVA) between March 2014 and May 2021. Molecular characterization of Plasmodium falciparum pfk13 and pfmdr1 genes was performed. We analyzed 232 imported malaria cases. The majority (68.53%) of the patients came from Angola and only three patients travelled to a non-African country; one to Brazil and two to Indonesia. P. falciparum was diagnosed in 81.47% of the cases, P. malariae in 7.33%, P. ovale 6.47% and 1.72% carried P. vivax. No mutations were detected in pfk13. Regarding pfmdr1, the wild-type haplotype (N86/Y184/D1246) was also the most prevalent (64.71%) and N86/184F/D1246 was detected in 26.47% of the cases. The typical imported malaria case was middle-aged male, traveling from Angola, infected with P. falciparum carrying wild type pfmdr1 and pfk13. Our study highlights the need for constant surveillance of malaria parasites imported into Portugal as an important pillar of public health.

scholarly journals Influx of diverse, drug resistant and transmissible Plasmodium falciparum into a malaria-free setting in Gulf Cooperation (GCC) countries.

2020 ◽  
Author(s):  
Abir Al-Rumhi ◽  
Zainab Al-Hashami ◽  
Salama Al-Hamidhi ◽  
Amal Gadalla ◽  
Raeece Naeem ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Plasmodium Falciparum ◽  
Resistance Genes ◽  
Multiple Drug Resistance ◽  
Marker Gene ◽  
Imported Malaria ◽  
Drug Resistant ◽  
Gametocyte Density ◽  
Gcc Countries ◽  
The Impact

Abstract Background Successful malaria control programs have interrupted local malaria transmission in almost all the Gulf Cooperation Council (GCC) countries. However, a massive influx of imported malaria via migrant workers from endemic areas sustains a threat for the re-introduction of local transmission. Here we examined the origin of imported malaria into one of the GCC countries (Qatar) and assessed the extent of genetic diversity, and carriage of drug resistance genes of imported Plasmodium falciparum and it’s potential to re-introduce the disease. Methods We examined imported malaria reported in Qatar, between 2013 and 2016. We focused on P. falciparum infections and estimated total parasite and gametocyte density using qPCR and qRT-PCR, respectively. In addition, we examined ten neutral microsatellites and four drug resistance genes, Pfmrp1, Pfcrt, Pfmdr1 and Pfkelch13 , to assess the extent of diversity of imported P. falciparum and its potential carriage of drug resistance genotypes respectively. Results The majority of imported malaria comprised P. vivax , while P. falciparum and mixed species infections ( P. falciparum /P. vivax ) were less frequent. The main origin of P. vivax was the Indian subcontinent, while P. falciparum was most apparent among expatriates from Africa. Imported P. falciparum was highly diverse carrying multiple genotypes as well as early and late gametocytes. We observed a high prevalence of SNPs implicated in drug resistance among imported P. falciparum , with some novel SNPs in Pfkelch13 . Conclusions The high influx of genetically diverse P. falciparum, with multiple drug resistance marker gene mutations and high capacity of producing gametocytes, sustains threat for re-introduction of drug resistant malaria into GCC countries. This scenario highlights the impact of current globalisation of movement of humans in reintroducing malaria infections to areas targeted for elimination.

scholarly journals Influx of diverse, drug resistant and transmissible Plasmodium falciparum into a malaria-free setting in Gulf Cooperation (GCC) countries.

2020 ◽  
Author(s):  
Abir Al-Rumhi ◽  
Zainab Al-Hashami ◽  
Salama Al-Hamidhi ◽  
Amal Gadalla ◽  
Raeece Naeem ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Plasmodium Falciparum ◽  
Resistance Genes ◽  
Multiple Drug Resistance ◽  
Marker Gene ◽  
Imported Malaria ◽  
Drug Resistant ◽  
Gametocyte Density ◽  
Gcc Countries ◽  
The Impact

Abstract Background Successful malaria control programs have interrupted local malaria transmission in almost all the Gulf Cooperation Council (GCC) countries. However, a massive influx of imported malaria via migrant workers from endemic areas sustains a threat for the re-introduction of local transmission. Here we examined the origin of imported malaria into one of the GCC countries (Qatar) and assessed the extent of genetic diversity, and carriage of drug resistance genes of imported Plasmodium falciparum and it’s potential to re-introduce the disease. Methods We examined imported malaria reported in Qatar, between 2013 and 2016. We focused on P. falciparum infections and estimated total parasite and gametocyte density using qPCR and qRT-PCR, respectively. In addition, we examined ten neutral microsatellites and four drug resistance genes, Pfmrp1, Pfcrt, Pfmdr1 and Pfkelch13 , to assess the extent of diversity of imported P. falciparum and its potential carriage of drug resistance genotypes respectively. Results The majority of imported malaria comprised P. vivax , while P. falciparum and mixed species infections ( P. falciparum /P. vivax ) were less frequent. The main origin of P. vivax was the Indian subcontinent, while P. falciparum was most apparent among expatriates from Africa. Imported P. falciparum was highly diverse carrying multiple genotypes as well as early and late gametocytes. We observed a high prevalence of SNPs implicated in drug resistance among imported P. falciparum , with some novel SNPs in Pfkelch13 . Conclusions The high influx of genetically diverse P. falciparum, with multiple drug resistance marker gene mutations and high capacity of producing gametocytes, sustains threat for re-introduction of drug resistant malaria into GCC countries. This scenario highlights the impact of current globalisation of movement of humans in reintroducing malaria infections to areas targeted for elimination.

scholarly journals Molecular Characterization of the CytochromebGene andIn VitroAtovaquone Susceptibility of Plasmodium falciparum Isolates from Kenya

2015 ◽  
Vol 59 (3) ◽  
pp. 1818-1821 ◽  
Author(s):  
Luicer A. Ingasia ◽  
Hoseah M. Akala ◽  
Mabel O. Imbuga ◽  
Benjamin H. Opot ◽  
Fredrick L. Eyase ◽  
...  
Keyword(s):  
Plasmodium Falciparum ◽  
Genetic Polymorphism ◽  
Molecular Characterization ◽  
Mutant Allele ◽  
Inhibitory Concentration ◽  
Wild Type Allele ◽  
Wild Type ◽  
Western Kenya ◽  
Content Type

ABSTRACTThe prevalence of a genetic polymorphism(s) at codon 268 in the cytochromebgene, which is associated with failure of atovaquone-proguanil treatment, was analyzed in 227Plasmodium falciparumparasites from western Kenya. The prevalence of the wild-type allele was 63%, and that of the Y268S (denoting a Y-to-S change at position 268) mutant allele was 2%. There were no pure Y268C or Y268N mutant alleles, only mixtures of a mutant allele(s) with the wild type. There was a correlation between parasite 50% inhibitory concentration (IC50) and parasite genetic polymorphism; mutant alleles had higher IC50s than the wild type.

scholarly journals The Antibody Response to Plasmodium falciparum: Cues for Vaccine Design and the Discovery of Receptor-Based Antibodies

2019 ◽  
Vol 37 (1) ◽  
pp. 225-246 ◽  
Author(s):  
Joshua Tan ◽  
Luca Piccoli ◽  
Antonio Lanzavecchia
Keyword(s):  
Public Health ◽  
Plasmodium Falciparum ◽  
Immune System ◽  
Antibody Response ◽  
Public Health Problem ◽  
Vaccine Design ◽  
Immunoglobulin Genes ◽  
And Control ◽  
Sterilizing Immunity

Plasmodium falciparum remains a serious public health problem and a continuous challenge for the immune system due to the complexity and diversity of the pathogen. Recent advances from several laboratories in the characterization of the antibody response to the parasite have led to the identification of critical targets for protection and revealed a new mechanism of diversification based on the insertion of host receptors into immunoglobulin genes, leading to the production of receptor-based antibodies. These advances have opened new possibilities for vaccine design and passive antibody therapies to provide sterilizing immunity and control blood-stage parasites.

scholarly journals Characterization of Plasmodium falciparum NEDD8 and identification of cullins as its substrates

2020 ◽  
Author(s):  
Manish Bhattacharjee ◽  
Navin Adhikari ◽  
Renu Sudhakar ◽  
Zeba Rizvi ◽  
Divya Das ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Plasmodium Falciparum ◽  
Western Blot ◽  
Wild Type ◽  
Malaria Parasites ◽  
Post Translational Modifications ◽  
Functional Conservation ◽  
Cellular Processes ◽  
Physiological Substrates

ABSTRACTA variety of post-translational modifications of Plasmodium falciparum proteins, including phosphorylation and ubiquitination, are shown to have key regulatory roles. The neural precursor cell expressed developmentally downregulated protein 8 (NEDD8) is a ubiquitin-like modifier of cullin-RING E3 ubiquitin ligases, which regulate diverse cellular processes, including the cell-cycle. Although neddylation pathway is conserved in eukaryotes, it is yet to be characterized in Plasmodium and related apicomplexan parasites. Towards studying the neddylation pathway in malaria parasites, we characterized P. falciparum NEDD8 (PfNEDD8) and identified cullins as its physiological substrates. PfNEDD8 is a 76 amino acid residue protein without the C-terminal tail, indicating that it can be readily conjugated. The wild type and mutant (Gly75Gly76 mutated to Ala75Ala76) PfNEDD8 were expressed in P. falciparum. Western blot of wild type PfNEDD8-expressing parasites indicated multiple high molecular weight conjugates, which were absent in the parasites expressing the mutant, indicating conjugation of NEDD8 to proteins through Gly76. Immunoprecipitation followed by mass spectrometry of wild type PfNEDD8-expressing parasites identified several proteins, including two putative cullins. Furthermore, we expressed PfNEDD8 in mutant S. cerevisiae strains that lacked endogenous NEDD8 (Δrub1) or NEDD8 conjugating E2 enzyme (ΔUbc12). The western blot of complemented strains and mass spectrometry of PfNEDD8 immunoprecipitate showed conjugation of PfNEDD8 to S. cerevisiae cullin cdc53, demonstrating functional conservation and cullins as the physiological substrates of PfNEDD8. The characterization of PfNEDD8 and identification of cullins as its substrates make ground for investigation of specific roles and drug target potential of neddylation pathway in malaria parasites.

scholarly journals Functional characterization of a glycine 185-to-valine substitution in human P-glycoprotein by using a vaccinia-based transient expression system.

1996 ◽  
Vol 7 (10) ◽  
pp. 1485-1498 ◽  
Author(s):  
M Ramachandra ◽  
S V Ambudkar ◽  
M M Gottesman ◽  
I Pastan ◽  
C A Hrycyna
Keyword(s):  
Drug Resistance ◽  
Atpase Activity ◽  
Cyclosporin A ◽  
Transient Expression ◽  
Expression System ◽  
Rhodamine 123 ◽  
Wild Type ◽  
P Glycoprotein ◽  
Transient Expression System

Human P-glycoprotein (Pgp) is a 170-kDa plasma membrane protein that confers multidrug resistance to otherwise sensitive cells. A mutation in Pgp, G185-->V, originally identified as a spontaneous mutation, was shown previously to alter the drug resistance profiles in cell lines that are stably transfected with the mutant MDR1 cDNA and selected with cytotoxic agents. To understand the mechanism by which the V185 mutation leads to an altered drug resistance profile, we used a transient expression system that eliminates the need for drug selection to attain high expression levels and allows for the rapid characterization of many aspects of Pgp function and biosynthesis. The mutant and wild-type proteins were expressed at similar levels after 24-48 h in human osteosarcoma (HOS) cells by infection with a recombinant vaccinia virus encoding T7 RNA polymerase and simultaneous transfection with a plasmid containing MDR1 cDNA controlled by the T7 promoter. For both mutant and wild-type proteins, photolabeling with [3H]azidopine and [125I]iodoarylazidoprazosin, drug-stimulated ATPase activity, efflux of rhodamine 123, and accumulation of radiolabeled vinblastine and colchicine were evaluated. In crude membrane preparations from HOS cells, a higher level of basal Pgp-ATPase activity was observed for the V185 variant than for the wild-type, suggesting partial uncoupling of drug-dependent ATP hydrolysis by the mutant. Several compounds, including verapamil, nicardipine, tetraphenylphosphonium, and prazosin, stimulated ATPase activities of both the wild-type and mutant similarly, whereas cyclosporin A inhibited the ATPase activity of the mutant more efficiently than that of the wild-type. This latter observation explains the enhanced potency of cyclosporin A as an inhibitor of the mutant Pgp. No differences were seen in verapamil-inhibited rhodamine 123 efflux, but the rate of accumulation was slower for colchicine and faster for vinblastine in cells expressing the mutant protein, as compared with those expressing wild-type Pgp. We conclude that the G185-->V mutation confers pleiotropic alterations on Pgp, including an altered basal ATPase activity and altered interaction with substrates and the inhibitor cyclosporin A.

scholarly journals Mapping the malaria parasite drug-able genome using in vitro evolution and chemogenomics

2017 ◽  
Author(s):  
Annie N. Cowell ◽  
Eva S. Istvan ◽  
Amanda K. Lukens ◽  
Maria G. Gomez-Lorenzo ◽  
Manu Vanaerschot ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Plasmodium Falciparum ◽  
Multidrug Resistance ◽  
Resistance Genes ◽  
Genome Analysis ◽  
Drug Targets ◽  
Malaria Parasite ◽  
Resistance Mechanisms ◽  
Whole Genome

AbstractChemogenetic characterization through in vitro evolution combined with whole genome analysis is a powerful tool to discover novel antimalarial drug targets and identify drug resistance genes. Our comprehensive genome analysis of 262 Plasmodium falciparum parasites treated with 37 diverse compounds reveals how the parasite evolves to evade the action of small molecule growth inhibitors. This detailed data set revealed 159 gene amplifications and 148 nonsynonymous changes in 83 genes which developed during resistance acquisition. Using a new algorithm, we show that gene amplifications contribute to 1/3 of drug resistance acquisition events. In addition to confirming known multidrug resistance mechanisms, we discovered novel multidrug resistance genes. Furthermore, we identified promising new drug target-inhibitor pairs to advance the malaria elimination campaign, including: thymidylate synthase and a benzoquinazolinone, farnesyltransferase and a pyrimidinedione, and a dipeptidylpeptidase and an arylurea. This deep exploration of the P. falciparum resistome and drug-able genome will guide future drug discovery and structural biology efforts, while also advancing our understanding of resistance mechanisms of the deadliest malaria parasite.One Sentence SummaryWhole genome sequencing reveals how Plasmodium falciparum evolves resistance to diverse compounds and identifies new antimalarial drug targets.

scholarly journals Molecular Surveillance of Antimalarial Resistance Pfcrt, Pfmdr1 and Pfk13 Polymorphisms in African Plasmodium Falciparum Imported Parasites to Wuhan, China

2020 ◽  
Author(s):  
Cheng Weijia ◽  
Xiaonan Song ◽  
Huabing Tan ◽  
Kai Wu ◽  
Jian Li
Keyword(s):  
Drug Resistance ◽  
Plasmodium Falciparum ◽  
Mixed Type ◽  
Antimalarial Drugs ◽  
Wild Type ◽  
Mutant Type ◽  
Molecular Surveillance ◽  
Severe Problem ◽  
Antimalarial Resistance ◽  
Effective Drugs

Abstract Background: The development of drug resistance in Plasmodium falciparum becomes a severe problem for malaria control globally. Before finding a practical solution, monitoring the susceptibility of P. falciparum resistance-related genes is crucial. It will offer valuable information on the drug resistance in malaria-endemic areas and guides the rational clinical use of antimalarial drugs.Methods：Filter paper blood was taken from patients with positive P. falciparum during 2017-2019 in Wuhan, China. The target fragments from pfcrt, pfmdr1, and k13 propeller (pfk13) genes of P. falciparum were amplified and sequenced. Subsequently, the polymorphisms of pfcrt, pfmdr1, and pfk13 and the haplotypes of Pfcrt and Pfmdr1 were analyzed.Results: Totally, 106 samples were collected. Subsequently, 98.11% (104/106), 100% (106/106), and 86.79% (92/106) of these samples were successfully amplified and sequenced for the pfcrt, pfmdr1, and pfk13 genes, respectively. The prevalence of Pfcrt K76T, Pfmdr1 N86Y, and Pfmdr1 Y184F mutation were 9.62%, 4.72%, and 47.17%, respectively. At codons 72-76 of pfcrt gene locus were showed three haplotypes, including CVMNK (wild-type), CVIET (mutation type), CV M/I N/E K/T (mixed type), with 87.50%, 9.62%, and 2.88% prevalence, respectively. For the pfmdr1 gene, including NY (wild type), NF and YF (mutant type), N Y/F, Y Y/F, and N/Y Y/F (mixed type), accounted for 34.91%, 43.40%, 3.77%, 15.09%, 0.94%, and 1.89%, respectively. A total of eleven Pfcrt/Pfmdr1 combined haplotypes, including six types of combined haplotypes, and five combined haplotypes with mixed-type, For pfk13, no mutation was detected. Conclusions: The wild-type SNPs and haplotypes for the pfcrt, and pfmdr1 genes become predominant in the current study. It indicates these isolates entirely or partly recovery their susceptibility to antimalarial drugs, including chloroquine, amodiaquine, and mefloquine. Moreover, it demonstrates these drugs can continue to be effective drugs for P. falciparum malaria cases treatment in Africa. Although no mutation is detected in pfk13, continuous molecular surveillance is still urgently necessary.

scholarly journals Role ofPfmdr1inIn VitroPlasmodium falciparum Susceptibility to Chloroquine, Quinine, Monodesethylamodiaquine, Mefloquine, Lumefantrine, and Dihydroartemisinin

2014 ◽  
Vol 58 (12) ◽  
pp. 7032-7040 ◽  
Author(s):  
Nathalie Wurtz ◽  
Bécaye Fall ◽  
Aurélie Pascual ◽  
Mansour Fall ◽  
Eric Baret ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Plasmodium Falciparum ◽  
Multidrug Resistance ◽  
Antimalarial Drug ◽  
Antimalarial Drugs ◽  
Wild Type ◽  
Antimalarial Drug Resistance ◽  
Content Type ◽  
Increased Susceptibility

ABSTRACTThe involvement ofPfmdr1(Plasmodium falciparummultidrug resistance 1) polymorphisms in antimalarial drug resistance is still debated. Here, we evaluate the association between polymorphisms inPfmdr1(N86Y, Y184F, S1034C, N1042D, and D1246Y) andPfcrt(K76T) andin vitroresponses to chloroquine (CQ), mefloquine (MQ), lumefantrine (LMF), quinine (QN), monodesethylamodiaquine (MDAQ), and dihydroartemisinin (DHA) in 174Plasmodium falciparumisolates from Dakar, Senegal. ThePfmdr186Y mutation was identified in 14.9% of the samples, and the 184F mutation was identified in 71.8% of the isolates. No 1034C, 1042N, or 1246Y mutations were detected. ThePfmdr186Y mutation was significantly associated with increased susceptibility to MDAQ (P= 0.0023), LMF (P= 0.0001), DHA (P= 0.0387), and MQ (P= 0.00002). The N86Y mutation was not associated with CQ (P= 0.214) or QN (P= 0.287) responses. ThePfmdr1184F mutation was not associated with various susceptibility responses to the 6 antimalarial drugs (P= 0.168 for CQ, 0.778 for MDAQ, 0.324 for LMF, 0.961 for DHA, 0.084 for QN, and 0.298 for MQ). ThePfmdr186Y-Y184 haplotype was significantly associated with increased susceptibility to MDAQ (P= 0.0136), LMF (P= 0.0019), and MQ (P= 0.0001). The additionalPfmdr186Y mutation increased significantly thein vitrosusceptibility to MDAQ (P< 0.0001), LMF (P< 0.0001), MQ (P< 0.0001), and QN (P= 0.0026) in wild-typePfcrtK76 parasites. The additionalPfmdr186Y mutation significantly increased thein vitrosusceptibility to CQ (P= 0.0179) inPfcrt76T CQ-resistant parasites.

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