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[In vivo chemosensitivity of Plasmodium falciparum to chloroquine in Burkina Faso: development of resistance 1988-1990].

A study of Plasmodium falciparum sensitivity to chloroquine was carried out in 1988 and 1990 in 5 localities, representatives of different climatic areas of Burkina Faso. The 7-day in vivo standard test performed in 1988 showed a total clearance failure of 25%. No significant difference with 1990 data was found, except for an increase of the resistance in the area of Fada N'Gourma, close to the border with Benin, Niger and Togo.

Animals↗

Malaria transmission-blocking activity in the plasma of Plasmodium falciparum gametocyte carriers in Cameroon.

Experimental infections of Anopheles gambiae were carried out with Plasmodium falciparum gametocytes from 65 naturally infected patients in Cameroon. A comparison was made between infections with blood containing autologous plasma and blood in which the plasma was replaced by plasma from donors without previous malaria exposure. A lower mosquito-infection rate was observed in 50 out of 65 autologous plasma samples. The transmission was completely blocked in 8 infections, whilst belonging exposures to heterologous plasma led to infected mosquitoes. Evidence is shown that blood plasma factors of gametocyte carriers from a population living in a malaria-endemic area are able to reduce transmission capacity.

Adolescent↗

In vitro activity and interaction of clindamycin combined with dihydroartemisinin against Plasmodium falciparum.

Combination regimens are considered a valuable tool for the fight against drug-resistant falciparum malaria. This study was conducted to evaluate the antimalarial potential of clindamycin in combination with dihydroartemisinin in continuously cultured and in freshly isolated Plasmodium falciparum parasites, measuring the inhibition of Plasmodium falciparum histidine-rich protein II synthesis. Interaction analysis revealed a synergistic or additive mode of interaction at various concentration ratios in all continuously cultured parasites at the 50% effective concentration (EC(50)) level. Antagonism was not found for any of the culture-adapted parasites. In fresh P. falciparum isolates, a fixed clindamycin-dihydroartemisinin combination exhibited additive activity at the EC(50) and EC(90) levels. The drug mixture showed no significant activity correlation to other commonly used antimalarials. The clindamycin-dihydroartemisinin combination appears to be a promising candidate for clinical investigation.

Animals↗

A bedside dipstick method to detect Plasmodium falciparum.

We conducted this study to determine efficacy of Parasight-F (an HRP-II antigen dipstick method to detect P. Falciparum) in children. A total of 30 children were enrolled in the age group of 2 months to 12 years whose peripheral smear showed asexual forms of Plasmodium falciparum. All patients were tested for presence of HRP-II antigen of Plasmodium falciparum in their blood by the Parasight-F dipstick test by either an EDTA sample or a finger prick blood sample. The sensitivity of Parasight-F was 83.3 % However, the sensitivity of Parasight-F to detect Plasmodium Falciparum in case of mixed Plasmodium (Vivax + Falciparum) infection was only 25 %. Also, all patients less than 6 months of age had a negative Parasight-F test. Parasitic index, prior treatment with antimalarials or severity of Falciparum malaria have no effect on the sensitivity of Parasight-F test. We conclude that Parasight-F is an effective tool for diagnosis of Plasmoduim falciparum malaria in children.

Animals↗

Antibody reactivity to conserved linear epitopes of Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1).

The Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1) family of protein antigens are involved in adhesion of P. falciparum infected erythrocytes to the capillary endothelium of the host. Antibodies to variable regions of these proteins, measured by agglutination, correlates with clinical protection against falciparum malaria. In this study we investigated the occurrence of antibodies to conserved sequences of these very variable proteins in a population living in an area endemic for falciparum malaria. Using the ELISA method, we were able to measure an antibody response to three synthetic peptides derived from conserved regions of PfEMP1. The antibody responses to these peptides increased with age and were higher in individuals with asymptomatic P. falciparum infection compared to individuals presenting with fever attributable to falciparum malaria. This indicates that antibodies recognising the conserved regions of PfEMP1 arise upon exposure to the parasite, and that these may be involved in the development of protection against malaria. Antibodies to the Pfalhesin peptide of the human aniontransporter, band3, were measured by the same method. The magnitude of this antibody response did not correlate with neither age nor clinical protection.

Amino Acid Sequence↗

Mathematical modelling of the within-host dynamics of Plasmodium falciparum.

The development of malaria due to Plasmodium falciparum is a complex, multi-stage process. It is usually characterized by an exponential growth in the number of parasite-infected erythrocytes, followed by marked oscillations in this number with a period of 48 h, which are eventually dampened. This course of events has been the subject of various mathematical models. In this paper we propose a new mathematical model for the in-host asexual erythrocytic development of P. falciparum malaria. Synchronicity of the infection is shown to be an inherent feature of infection, irrespective of the duration of merozoite release from the liver. It will, therefore, cause periodic symptoms, as known in malaria patients. We also simulate the effects of an induced host immune response and show how the level of immunity affects the development of disease. The simulations fit well with the clinical observations. We show how infection can become asynchronous and discuss the effect of desynchronization on the circulating and total parasitaemia and demonstrate that synchronized broods will show parasitaemia fluctuations.

Animals↗

Comparison of PCR-RFLP and Genescan-based genotyping for analyzing infection dynamics of Plasmodium falciparum.

Parameters describing the infection dynamics of Plasmodium falciparum are important determinants of the potential impact of interventions and are potential outcome measurements for malaria intervention trials. Low parasite densities, periodic sequestration of parasites, and the presence of multiple concurrent infections make it essential to use molecular techniques to estimate the force of infection and duration of infections in endemic areas. We now compare two approaches for tracking individual genotypes of the highly polymorphic merozoite surface protein 2: 1) fluorescence-labeled polymerase chain reaction (PCR) and GeneScan-sizing and 2) restriction fragment length polymorphism (RFLP). We analyze samples from a longitudinal field study in Ghana and use statistical approaches that allow for imperfect detectability. The two methods gave broadly similar estimates of parasite dynamics, but GeneScan is more precise and can achieve a higher throughput. The analysis of parasite dynamics indicated an average duration of infection of 210 days by GeneScan versus 152 days by PCR-RFLP in the study population in Kassena-Nankana, Northern Ghana. This reflects the good performance of GeneScan-based genotyping for studies of parasite infection dynamics.

Adolescent↗

Chloroquine resistant Plasmodium falciparum infection from Lampung and South Sumatra, Indonesia.

A report was made of 4 cases of chloroquine resistant Plasmodium falciparum infections. The infections, detected in Jakarta, were imported from Kotabumi, Tanjung Karang, the Island of Pidada in the Lampung Province and from Pangkalpinang on the Island Bangka in the Province of South Sumatra. Treatment with courses of 1500 mg chloroquine base and with increased dosages up to 2250 mg base failed to cure the patients. The chloroquine sensitivity test in vitro was carried out in 3 patients, which showed that the Plasmodium falciparum strains were resistant to chloroquine at the R I level. The strains appeared to be similar to the Malaya Camp strain. In vivo observations revealed that the parasites were resistant at the R I level with a delayed recrudescence. The chloroquine resistant falciparum malaria cases, acquired in South Sumatra, may therefore be regarded as the first reported cases from a focus outside the already known two foci in Indonesia, namely East Kalimantan and Irian Jaya. It may be expected that chloroquine resistant Plasmodium falciparum will be encountered in other parts of Indonesia in the near future. The use of a combination of sulfadoxine and pyrimethamine should not be recommended in Indonesia because chloroquine is still considered the drug of choice against all malaria infections in Indonesia.

Adult↗

Molecular mechanism of host specificity in Plasmodium falciparum infection: role of circumsporozoite protein.

Plasmodium falciparum sporozoites invade liver cells in humans and set the stage for malaria infection. Circumsporozoite protein (CSP), a predominant surface antigen on sporozoite surface, has been associated with the binding and invasion of liver cells by the sporozoites. Although CSP across the Plasmodium genus has homology and conserved structural organization, infection of a non-natural host by a species is rare. We investigated the role of CSP in providing the host specificity in P. falciparum infection. CSP from P. falciparum, P. gallinaceum, P. knowlesi, and P. yoelii species representing human, avian, simian, and rodent malaria species were recombinantly expressed, and the proteins were purified to homogeneity. The recombinant proteins were evaluated for their capacity to bind to human liver cell line HepG2 and to prevent P. falciparum sporozoites from invading these cells. The proteins showed significant differences in the binding and sporozoite invasion inhibition activity. Differences among proteins directly correlate with changes in the binding affinity to the sporozoite receptor on liver cells. P. knowlesi CSP (PkCSP) and P. yoelii CSP (PyCSP) had 4,790- and 17,800-fold lower affinity for heparin in comparison to P. falciparum CSP (PfCSP). We suggest that a difference in the binding affinity for the liver cell receptor is a mechanism involved in maintaining the host specificity by the malaria parasite.

Animals↗

[Drug resistance of Plasmodium falciparum malaria in Equatorial Africa. Apropos of cases observed at a military unit stationed in Gabon].

The existence of Plasmodium falciparum chloroquine-resisting clones in equatorial Africa is quite a recent event. Among a French military unit of 138 people who had lived in Gabon from April to August 1986, 24 of them have caught malaria due to Plasmodium falciparum. This allows us to perceive the impact of this event because of the perfect control of chloroquine prophylaxis and the good knowledge of all the attacks of fever which occurred in the field. Admitting 10 of those people into the department of internal medicine of the Military Hospital Desgenettes located in Lyons, we could have seen that trophozoites of Plasmodium falciparum were still persisting 2 months after their return for 8 of them in spite of carrying on chemoprophylaxis. Observing that in the field, sets the problem of the chemoprophylaxis of malaria for the troops maneuvering in chloroquine-resisting areas and warrants, if justified, a work protocol aimed at solving it.

Adult↗

Compliance with a 2 day course of artemether-mefloquine in an area of highly multi-drug resistant Plasmodium falciparum malaria.

AIMS: Multi-drug resistant Plasmodium falciparum malaria is a rapidly increasing problem in the world, particularly Thailand. Practical antimalarial regimens which are highly effective against multi-drug resistant parasites with short-term course of administration are needed. In this study, we assessed the patient compliance of a short course regimen using artemether-mefloquine. METHODS: Clinical effectiveness (efficacy, tolerability and patient compliance) of a 2-day regimen of artemether-mefloquine was evaluated in 126 patients with acute uncomplicated falciparum malaria who were attending the two malaria clinics in an area of highly multi-drug resistant P. falciparum malaria (Thai-Myanmar border). Patients were treated with a single oral dose of 300 mg artemether on the day of attendance. Two additional doses of mefloquine were given for home treatment on the following day (750 and 500 mg after breakfast and lunch, respectively). RESULTS: The combination regimen was effective, with a cure rate of 92.6%. Based upon the concentrations of whole blood mefloquine on day-2, compliance for this 2 day regimen of artemether-mefloquine was 98.1% (full compliance 86.8%, partial compliance 11.3%, non-compliance 1.9%). CONCLUSIONS: We conclude that the 2 day regimen of artemether-mefloquine is, at present, a good alternative regimen for the treatment of uncomplicated multi-drug resistant falciparum malaria.

Administration, Oral↗

Human malaria in immunocompromised mice: an in vivo model to study defense mechanisms against Plasmodium falciparum.

We have recently described that sustained Plasmodium falciparum growth could be obtained in immunodeficient mice. We now report the potential of this new mouse model by assaying the effect of the passive transfer of antibodies (Abs) which in humans have had a well-established effect.Our results show that the total African adult hyperimmune immunoglobulin Gs (HI-IgGs) strongly reduce P. falciparum parasitemia similarly to that reported in humans, but only when mice are concomitantly reconstituted with human monocytes (HuMNs). In contrast, neither HI-IgGs nor HuMNs alone had any direct effect upon parasitemia. We assessed the in vivo effect of epitope-specific human Abs affinity-purified on peptides derived either from the ring erythrocyte surface antigen (RESA) or the merozoite surface protein 3 (MSP3). The inoculation of low concentrations of anti-synthetic peptide from MSP3, but not of anti-RESA Abs, consistently suppressed P. falciparum in the presence of HuMNs. Parasitemia decrease was stronger and faster than that observed using HI-IgGs and as fast as that induced by chloroquine. Our observations demonstrate that this mouse model is of great value to evaluate the protective effect of different Abs with distinct specificity in the same animal, a step hardly accessible and therefore never performed before in humans.

Adult↗

Comparative genomics of transcriptional control in the human malaria parasite Plasmodium falciparum.

The life cycle of the parasite Plasmodium falciparum, responsible for the most deadly form of human malaria, requires specialized protein expression for survival in the mammalian host and insect vector. To identify components of processes controlling gene expression during its life cycle, the malarial genome--along with seven crown eukaryote group genomes--was queried with a reference set of transcription-associated proteins (TAPs). Following clustering on the basis of sequence similarity of the TAPs with their homologs, and together with hidden Markov model profile searches, 156 P. falciparum TAPs were identified. This represents about a third of the number of TAPs usually found in the genome of a free-living eukaryote. Furthermore, the P. falciparum genome appears to contain a low number of sequences, which are highly conserved and abundant within the kingdoms of free-living eukaryotes, that contribute to gene-specific transcriptional regulation. However, in comparison with these other eukaryotic genomes, the CCCH-type zinc finger (common in proteins modulating mRNA decay and translation rates) was found to be the most abundant in the P. falciparum genome. This observation, together with the paucity of malarial transcriptional regulators identified, suggests Plasmodium protein levels are primarily determined by posttranscriptional mechanisms.

Animals↗

Sensitivity to antimalarial drugs by Plasmodium falciparum in Goundry, Oubritenga province, Burkina Faso.

Plasmodium falciparum susceptibility to chloroquine in vivo and to chloroquine, mefloquine, quinine, amodiaquine and sulfadoxine/pyrimethamine in vitro was investigated in children living in Goundry village, Oubritenga Province (Burkina Faso) in November 1992. An extended WHO in vivo field test was used, with follow-up on days 2, 4, 7, 14, 21 and 28 after treatment with 25 mg chloroquine per kg body weight given over 3 days, in children from 2 to 8 years old with P. falciparum monospecific infection, asexual parasitaemia > 800 parasites/microliter of blood and negative Bergqvist urine tests. At the same time, the in vitro response was assessed using WHO standard test kits. Out of the 71 in vivo responses examined, 50 (70.4%) were classified as resistant to chloroquine at RI (43.6%) or RII (26.8%) levels. There were no RIII responses. Out of the 43 isolates tested for chloroquine susceptibility in vitro, 32 (74.4%) were resistant to the drug with mean EC50 and EC99 values of 1.41 mumol and 10.96 mumol/l of blood, respectively. Resistance to sulfadoxine/pyrimethamine in vitro was observed in one out of 19 tested cases, with mean EC50 and EC99 values of 0.00002 mumol and 35.05 mumol/l of blood, respectively. All isolates were inhibited by mefloquine at 12.8 mumol/l of blood, quinine at 51.2 mumol/l of blood and amodiaquine at 0.4 mumol/l of blood, indicating full sensitivity to these 3 drugs. The present study demonstrates the high prevalence of chloroquine-resistant strains of P. falciparum in the study area of Burkina Faso and indicates that isolates resistant to sulfadoxine/pyrimethamine may also be present.

Amodiaquine↗

[Recombination and cloning of MSP1(19) and PfCMR of Plasmodium falciparum].

AIM: To construct a recombinant plasmid DNA encoding multiantigens of Plasmodium falciparum and to provide the requirements for DNA immunization. METHODS: Two oligonucleotide primers were designed to amplify MSP1(19), the purified PCR products were digested by Sal I + Xba I, and the recombinant plasmid pWR450-1/PfCMR was digested by EcoR I + Sal I to recover PfCMR gene. PfCMR and MSP1(19) gene fragments were linked and recombined with mammalian expression vector pcDNA3. RESULTS: The MSP1(19) gene fragment with about 363 base pairs were specifically amplified by using PCR technique. The positive recombinant pcDNA3-PfCMR-MSP1(19) (named pcDNA3-Pf8) was screened and identified by agarose gel electrophoresis, endonulease digestion and PCR technique, the whole length of Pf8 is 618 bp. CONCLUSION: By specifically amplifying MSP1(19) gene at the C-terminal of MSP1, a recombinant plasmid pcDNA3-Pf8 encoding multiantigens of Plasmodium falciparum was successfully constructed.

Animals↗

In vitro activity of quinolines against Plasmodium falciparum in Gabon.

The assessment of drug sensitivity of Plasmodium falciparum to antimalarial drugs is of vital interest for malaria endemic regions. We conducted a follow-up study to monitor the in vitro activity of the most commonly used quinolines against fresh P. falciparum isolates in Lambaréné, Gabon by measuring schizont maturation inhibition in 2002. Mean 50% effective concentration levels for chloroquine, quinine, and mefloquine were 5.5micromol/l blood, 286nmol/l blood medium mixture (BMM), and 1.1micromol/l blood, respectively. All isolates (n=40) were found to be highly resistant to chloroquine. One isolate was resistant to mefloquine and five isolates were presenting borderline-resistance. All isolates were inhibited by quinine concentrations below the threshold of resistance (n=43). Besides the observation of an increasing number of borderline resistant isolates to mefloquine, an extremly high parasite resistance to chloroquine-still officially the first line antimalarial in Gabon-seems to be of particular concern.

Adolescent↗

Molecular markers for chloroquine-resistant Plasmodium falciparum malaria in Thailand and Laos.

Chloroquine-resistant Plasmodium falciparum is well documented in Thailand. Laos, however, continues to use chloroquine (CQ) as the first-line therapy for the treatment of P. falciparum malaria. The objective of the present study was to determine the prevalence, in these two areas, of the cg2, pfmdr1 and pfcrt allelic types that have previously been associated with CQ resistance. Isolates of P. falciparum were collected from participants in ongoing treatment studies conducted in Thailand (near the Thai-Cambodian border) and in Laos (Vang Vieng district). The pfmdr1 and pfcrt alleles were characterized by PCR-RFLP and mutations in cg2 were characterized by PCR and single-stranded-conformation-polymorphism (SSCP) electrophoresis. Eight (32%) of the 25 Laotian isolates but only one (4%) of the 25 Thai isolates were found to contain the pfmdr1 mutation N86Y (P = 0.02). In contrast, the cg2 polymorphisms previously associated with CQ resistance were present in only 10 of the isolates from Laos but 24 of those from Thailand (40% v. 96%; P < 0.001). All the samples from both countries contained the pfcrt K76T mutant allele reported to confer resistance to CQ. The results may indicate that drug pressure for the maintenance of the pfmdr1 and cg2 alleles varies in intensity in the Thai and Laotian study areas, probably reflecting differences in the national malaria-treatment policies of Thailand and Laos.

ATP-Binding Cassette Transporters↗