A review of antimalarial plants used in traditional medicine in communities in Portuguese-Speaking countries: Brazil, Mozambique, Cape Verde, Guinea-Bissau, São Tomé and Príncipe and Angola

The isolation of bioactive compounds from medicinal plants, based on traditional use or ethnomedical data, is a highly promising potential approach for identifying new and effective antimalarial drug candidates. The purpose of this review was to create a compilation of the phytochemical studies on medicinal plants used to treat malaria in traditional medicine from the Community of Portuguese-Speaking Countries (CPSC): Angola, Brazil, Cape Verde, Guinea-Bissau, Mozambique and São Tomé and Príncipe. In addition, this review aimed to show that there are several medicinal plants popularly used in these countries for which few scientific studies are available. The primary approach compared the antimalarial activity of native species used in each country with its extracts, fractions and isolated substances. In this context, data shown here could be a tool to help researchers from these regions establish a scientific and technical network on the subject for the CPSC where malaria is a public health problem.

Plasmodium vivax in the Democratic Republic of East Timor: Parasite prevalence and antifolate resistance-associated mutations.

In the Democratic Republic of East Timor, Plasmodium falciparum and Plasmodium vivax malaria coexist, but limited information is available about the latter species. Consequently, the prevalence of P. vivax and of its corresponding antifolate resistance-associated mutations in the pvdhfr and pvdhps genes was assessed here. Blood samples were collected from 650 individuals distributed among six districts, over two different periods, by either passive case detection (PCD) or active case detection (ACD). As expected, malaria was over-represented in the PCD sample (26% PCD vs 5% ACD), because the infection increases medical care seeking. Additionally, the relative frequency of P. vivax infections in symptomatic individuals (37%) was twice as high as the one in the asymptomatic sampling group (18%), suggesting that that this parasite is accounting for a significant proportion malaria-attributed morbidity. The frequency of specific sulfadoxine-pyrimethamine resistance-associated mutations genes was ascertained in P. vivax positive samples by PCR-RFLP. Although no mutants were detected in codons 383 and 553 of pvdhps, 48%, 76% and 82% of P. vivax-infected samples harbored the dhfr 33L, 58R and 117N mutations, respectively. Additionally, the frequency of parasites carrying both pvdhfr 58R and 117N mutant alleles accounted for a third of all genotypes analyzed, most likely due to inadvertent SP use in the past. In conclusion, evidence-based information is provided to promote optimized drug deployment and limit the evolution of resistance to antifolate resistance in P. vivax from East Timor.

Resistência à sulfadoxina-pirimetamina em Maputo, Moçambique: presença de mutações nos genes dhfr e dhps do Plasmodium falciparum / Sulfadoxine-pyrimethamine resistance in Maputo, Mozambique: presence of mutations in the dhfr and dhps genes of Plasmodium falciparum

Foram analisadas a freqüência e distribuição de mutações nos genes dihidrofolato redutase e dihidropteroato sintetase do Plasmodium falciparum, usando a metodologia de reação em cadeia da polimerase e polimorfismos de hidrólise por enzimas de restrição, em amostras de sangue infectado proveniente de crianças moçambicanas, residentes em Maputo. A análise foi feita antes e 7 dias após o tratamento com sulfadoxina-pirimetamina (S/P). Os resultados mostraram a ocorrência de mutações pontuais nos genes estudados e a presença de combinações de três alelos em dhfr (51Ile, 59Arg e 108Asn) e do quintúplo mutante (dhfr 51Ile, 59Arg, 108Asn e dhps 437Gly, 540Glu), ambas situações associadas à falha terapêutica no sétimo dia após tratamento com S/P. Esses achados mostram a importância de se estudar a resistência à S/P em Moçambique, e como os marcadores moleculares de resistência aos antimaláricos podem fornecer dados importantes para a política nacional de controlo da malária.

The frequency and distribution of mutations in Plasmodium falciparum, dihydrofolate reductase and dihydropteroate synthase genes were analyzed, using the polymerase chain reaction and restriction fragment length polymorphism methodology, in infected blood samples from Mozambican children living in Maputo, before and seven days after treatment with sulfadoxine/pyrimethamine (S/P). The results showed the occurrence of point mutations in the genes studied and the presence of combinations of three alleles in dhfr (51Ile, 59Arg and 108Asn) and "quintuple" mutant (dhfr 51Ile, 59Arg, 108Asn and dhps 437Gly, 540Glu). Both of these situations were associated with seven-day therapeutic failure, following treatment with S/P. These findings show the importance of studying S/P resistance in Mozambique, and how molecular markers for antimalarial resistance can provide important data for national malaria control policy.

The control of an epidemic outbreak of Marburg hemorrhagic fever in Angola, 2004-2005 / No disponible

Entre Octubre del 2004 y Septiembre del 2005 Angola vivió un brote epidémico de fiebre hemorrágica debida al virus Marburg. La epidemia se dio por oficialmente por concluida el 7 de Noviembre del 2005, y se consideró la más grave de las ocurridas en todo el mundo, con 252 casos y 227 muertes, lo que suponía una tasa de mortalidad del 90%. Considerando que es un agente patógeno de gran virulencia para el cual no hay vacunas o tratamiento específico, estas epidemias precisan de una enorme cantidad de recursos humanos, financieros y técnicos tanto nacionales como internacionales Es importante poner de manifiesto que el laboratorio del Center for Diseases Control de Atlanta fue el responsable del primer aislamiento del virus y de su identificación como causante de la epidemia. Posteriormente fue el encargado de monitorizar la epidemia y estudiar todas las muestras biológicas recogidas de los casos sospechosos. Hasta este momento ha sido imposible identificar el foco primario de la enfermedad, aunque existen sugerencias a nivel local sobre la posibilidad de que el hospital provincial de UIge haya sido el sitio inicial y responsable de la diseminación de la epidemia a través de la infección nosocomial. En una segunda fase la ausencia de una adecuada educación para la salud, el pánico y las tradiciones culturales locales fueron los elementos cruciales para la diseminación de la enfermedad a otras provincias del país. En este contexto, la participación de sociólogos y antropólogos fue un factor clave para el control de las epidemias, en paralelo a las medidas de bioseguridad y detección activa emprendidas (AU)

Angola has lived an epidemic outbreak of hemorrhagic fever caused by Marburg virus between October 2004 and September 2005. The epidemic was officially terminated in November 7th 2005, and considered the most severe ever occurred in the world, with 252 cases and 227 deaths, with a lethality rate of 90%. Considering that it is a pathogenic agent of great virulence for which there are no vaccines or specific treatment, these epidemics caused a huge recruitment of national and international human, financial and technical resources despite some local constraints. It must be outlined that CDC-Atlanta laboratory was the responsible for the first isolation of the virus as the cause of the epidemics, and later on, for monitoring the epidemics and manipulating all biological samples collected from suspected patients. Until now it was impossible to identify the primary focus of the disease, though there are suggestions from local oral comments that Uige provincial hospital may have been the initial site and responsible for the dissemination of the epidemics through nosocomial infection. In a second phase, lack of proper health education, panic and local cultural traditions were the crucial elements for disease spread to other provinces of the country. In this context, the participation of sociologist and anthropologists was a key factor for the epidemics control, in parallel with the biosafety and active survey measures undertaken (AU)

Epidemiological characterization of Plasmodium falciparum in the Republic of Cabo Verde: implications for potential large-scale re-emergence of malaria.

BACKGROUND: Malaria has come near eradication at archipelago of Cabo Verde in 1970. Infections are now only observed in Santiago, where outbreaks occur. In these islands, malaria is considered by the international community as being of limited risk and, therefore, no prophylaxis is recommended. Since the understanding of factors that determine malaria outbreaks are crucial for controlling the disease, the present study aimed to investigate if the malaria infections observed in Santiago Island are maintained in isolated foci and in asymptomatic individuals. METHODS: The occurrence of asymptomatic carriers in villages with history of malaria as well as the level of exposure of these populations were investigated using PCR and serological analyses. RESULTS: Results indicate that malaria is maintained as asymptomatic and sub-patent infections and that the majority of the circulating parasite populations harbour chloroquine-resistant mutations. CONCLUSION: These observations highlight the alarming prospect of malaria to become a serious public health problem and underscore the need for a tighter surveillance.

Real-time quantitative PCR with SYBR Green I detection for estimating copy numbers of nine drug resistance candidate genes in Plasmodium falciparum.

BACKGROUND: Evaluating copy numbers of given genes in Plasmodium falciparum parasites is of major importance for laboratory-based studies or epidemiological surveys. For instance, pfmdr1 gene amplification has been associated with resistance to quinine derivatives and several genes involved in anti-oxidant defence may play an important role in resistance to antimalarial drugs, although their potential involvement has been overlooked. METHODS: The DeltaDeltaCt method of relative quantification using real-time quantitative PCR with SYBR Green I detection was adapted and optimized to estimate copy numbers of three genes previously indicated as putative candidates of resistance to quinolines and artemisinin derivatives: pfmdr1, pfatp6 (SERCA) and pftctp, and in six further genes involved in oxidative stress responses. RESULTS: Using carefully designed specific RT-qPCR oligonucleotides, the methods were optimized for each gene and validated by the accurate measure of previously known number of copies of the pfmdr1 gene in the laboratory reference strains P. falciparum 3D7 and Dd2. Subsequently, Standard Operating Procedures (SOPs) were developed to the remaining genes under study and successfully applied to DNA obtained from dried filter blood spots of field isolates of P. falciparum collected in São Tomé & Principe, West Africa. CONCLUSION: The SOPs reported here may be used as a high throughput tool to investigate the role of these drug resistance gene candidates in laboratory studies or large scale epidemiological surveys.

Studies in a co-infection murine model of Plasmodium chabaudi chabaudi and Leishmania infantum: interferon-gamma and interleukin-4 mRNA expression

This work aimed to study the T helper type 1/2 (Th1/Th2) cytokine profile in a co-infection murine model of Plasmodium chabaudi chabaudi and Leishmania infantum. Expression of interferon-gamma and interleukin-4 (IL-4) was analyzed, in spleen and liver of C57BL/6 mice, by reverse transcriptase-polymerase chain reaction. High levels of IFN-gamma expression did not prevent the progression of Leishmania in co-infected mice and Leishmania infection did not interfere with the Th1/Th2 switch necessary for Plasmodium control. The presence of IL-4 at day 28 in co-infected mice, essential for Plasmodium elimination, was probably a key factor on the exacerbation of the Leishmania infection.

Do bednets reduce malaria transmission by exophagic mosquitoes?

The impact of bednet coverage on malaria prevalence in a suburb of São Tomé was monitored, by passive case detection, over a three-year period (1997-1999), when bednet use increased from 20 to 74%. Malaria parasites were detected in 1651 (41.6%) of the 3967 slides taken during the study. All four human malaria parasites were seen, with Plasmodium falciparum being the predominant species (94.9% of positive slides). Prevalence of malaria among residents decreased, particularly in 1-4 year olds. In addition, there was a concomitant decrease in prevalence also among non-net users, suggesting a mass effect on transmission, even though the only vector in the area is largely exophagic and zoophilic.

Chromosome size variation occurs in cloned Plasmodium falciparum on in vitro cultivation

Variaçöes de tamanho nos cromossomos de linhagens de parasitos derivados de um clone de Plasmodium falciparum. Tais variaçöes desenvolveram-se durante períodos de muitos meses em parasitos haplóides na fase eritrocitária, mantidos in vitro. As variaçöes foram mais numerosas em populaçöes com resistência à mefloquina, induzida pelo cultivo sob pressäo de droga, do que em parasitas controles mantidos em meio de cultura sem droga. Em alguns casos, as modificaçöes de tamanho foram täo grandes que alteraram a ordem relativa das bandas de DNA cromossômico em separaçöes através de eletroforese em gel de "pulsed-field gradient". As regiöes subteloméricas dos cromossomos säo especialmente ativas, estando envolvidas em rearranjos responsáveis pelas alteraçöes de tamanho. O papel biológico destes rearrranjos näo é conhecido. Entretanto, sua ocorrência em experimentos controles, bem como sua frequente apariçäo em condiçöes de pressäo por mefloquina, sugerem que tais rearranjos reflititiam modificaçöes generalizadas da estrutura genômica em resposta às condiçöes ambientais