Design, synthesis, and characterization of novel aminoalcohol quinolines with strong in vitro antimalarial activity.

Malaria is the fifth most lethal parasitic infections in the world. Herein, five new series of aminoalcohol quinolines including fifty-two compounds were designed, synthesized and evaluated in vitro against Pf3D7 and PfW2 strains. Among them, fourteen displayed IC50 values below or near of 50.0 nM whatever the strain with selectivity index often superior to 100.17b was found as a promising antimalarial candidate with IC50 values of 14.9 nM and 11.0 nM against respectively Pf3D7 and PfW2 and a selectivity index higher than 770 whatever the cell line is. Further experiments were achieved to confirm the safety and to establish the preliminary ADMET profile of compound 17b before the in vivo study performed on a mouse model of P. berghei ANKA infection. The overall data of this study allowed to establish new structure-activity relationships and the development of novel agents with improved pharmacokinetic properties.

What could explain the late emergence of COVID-19 in Africa?

At the end of November 2019, a novel coronavirus responsible for respiratory tract infections emerged in China. Despite drastic containment measures, this virus, known as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), spread in Asia and Europe. The pandemic is ongoing with a particular hotspot in southern Europe and America in spring 2020. Many studies predicted an epidemic in Africa similar to that currently seen in Europe and the USA. However, reported data do not confirm these predictions. Several hypotheses that could explain the later emergence and spread of the coronavirus disease 2019 (COVID-19) pandemic in African countries are being discussed, including the lack of health-care infrastructure capable of clinically detecting and confirming COVID-19 cases, the implementation of social distancing and hygiene, international air traffic flows, the climate, the relatively young and rural population, the genetic polymorphism of the angiotensin-converting enzyme 2 receptor, cross-immunity and the use of antimalarial drugs.

Changes in malaria epidemiology in France and worldwide, 2000-2015.

In 2015, 212 million new cases of malaria were reported, causing 429,000 deaths. The World Health Organization (WHO) estimated a 41% decrease in the number of new cases worldwide between 2000 and 2015. The number of deaths from malaria fell by 62% worldwide and by 71% in Africa. In mainland France, malaria is mainly imported by travelers or migrants from endemic areas, in particular sub-Saharan Africa (95%). In France, the number of imported malaria cases, mainly due to Plasmodium falciparum (85%), was estimated at about 82,000 for the period 2000-2015. Over the same period, 6,468 cases of malaria were reported in the French armed forces, of which 2,430 cases (37.6%) were considered as imported because occurring outside of endemic areas. The number of malaria cases also fell between 2000 and 2015 in Mayotte and French Guiana, a malaria transmission zone. Mayotte has entered the elimination of malaria with less than 15 cases per year. In French Guiana, between 300 and 500 cases have been reported annually in recent years. The decline in morbidity and mortality is usually attributed to vector control measures and improved access to effective treatments. However, the Anopheles mosquitoes that transmit the disease have developed resistance against most insecticides. Similarly, malaria parasites have developed resistance against most of the antimalarial drugs used as prevention or treatment, even the latest marketed combinations such as artemisinin-based combination therapies.

Genetic diversity and deletion of Plasmodium falciparum histidine-rich protein 2 and 3: a threat to diagnosis of P. falciparum malaria.

BACKGROUND: PfHRP2-based rapid diagnostic tests (RDTs), based on the recognition of the Plasmodium falciparum histidine-rich protein 2, are currently the most used tests in malaria detection. Most of the antibodies used in RDTs also detect PfHRP3. However, false-negative results were reported. Significant variation in the pfhrp2 gene could lead to the expression of a modified protein that would no longer be recognized by the antibodies used in PfHRP2-based RDTs. Additionally, parasites lacking the PfHRP2 do not express the protein and are, therefore, not identifiable. AIMS: This review aims to assess the pfhrp2 and pfhrp3 genetic variation or the prevalence of gene deletion in areas where malaria is endemic and describe its implications on RDT use. SOURCES: Publications of interest were identified using PubMed, Google Scholar and Google. CONTENT: More than 18 types of amino acid repeats were identified from the PfHRP2 sequences. Sequencing analysis revealed high-level genetic variation in the pfhrp2 and pfhrp3 genes (>90% of variation in Madagascar, Nigeria or Senegal) both within and between countries. However, genetic variation of PfHRP2 and PfHRP3 does not seem to be a major cause of false-negative results. The countries that showed the highest proportions of pfhrp2-negative parasites were Peru (20%-100%) and Guyana (41%) in South America, Ghana (36%) and Rwanda (23%) in Africa. High prevalence of pfhrp2 deletion causes a high rate of false-negatives results. IMPLICATIONS: Presence of parasites lacking the pfhrp2 gene may pose a major threat to malaria control programmes because P. falciparum-infected individuals are not diagnosed and properly treated.

Contribution of the French army health service in support of expertise and research in infectiology in Africa.

Historically, infectious diseases have caused more casualties than battle. The French military health service therefore developed a range of research on vector-borne diseases such as malaria and arboviruses, antibiotic resistance, infectious agents that can be used as biological weapons and vaccines. The main objective is to control naturally acquired or provoked infectious diseases and limit their impact on armed forces as well as on civilian populations in France or abroad, particularly in Africa and anywhere French armies may be deployed. The expertise of the military health service teams in manipulating agents requiring high level of biosafety precautions and in organizing and providing medical care in unnatural conditions, including the battlefield, associated with complementarity staff experience (physicians, biologists, epidemiologists, researchers, pharmacists, logisticians), has been used in the management of the Ebola outbreak in Guinea.

Mobile populations across the Mediterranean Sea and beyond: travel medicine, mass gathering medicine and homeless health.

The Méditerranée Infection institute is internationally recognized for its expertise in infectious diseases and tropical medicine, and is one of the most active research centres for infectious diseases in Europe. Surveillance and research addressing infectious diseases in globally mobile populations is one of the strong components of the research conducted at the institute. A significant amount of clinical, microbiologic and epidemiologic works have been conducted in international travellers, pilgrims participating in large international religious gatherings, economic migrants and homeless migrant people over the last decades by our group. Our strong anchoring in several countries around the Mediterranean Sea and beyond, as well as the pivotal role of Marseille in the EuroTravNet and GeoSentinel international networks that monitor travel-associated diseases, reinforce our leading position in the fields of travel and tropical medicine, mass gathering medicine and homeless health.

[Role of primaquine in malaria control and elimination in French-speaking Africa]. / Place de la primaquine dans la lutte contre le paludisme en Afrique francophone.

Primaquine, an 8-aminoquinoline, is a relatively unknown and underutilized drug in French-speaking African countries. It acts against the liver stage parasites of all human malaria species, asexual blood stages of Plasmodium vivax and, to a lesser degree, Plasmodium falciparum; P. falciparum mature gametocytes, and P. vivax and Plasmodium ovale hypnozoites. Gastrointestinal disturbances are its most common side effects. The clinical utility of primaquine is limited due to its hematological side effects in individuals with glucose-6-phosphate dehydrogenase (G6PD) deficiency and other contraindications (pregnant woman, breastfeeding woman, infants less than 6 months old). In the light of the recent recommendations of the World Health Organization (WHO), we propose to examine how primaquine can be used in French-speaking Africa to improve malaria control and move towards malaria elimination. Two indications supported by the WHO are of relevance in Africa. First, artemisinin-based combination therapies and primaquine given as a single low dose (0.25 mg base/kg) are effective to kill asexual and sexual parasites of P. falciparum, are well-tolerated, and have very little risk even in mild to moderate G6PD-deficient patients. This strategy may be helpful to contain transmission in an area in Africa where P. falciparum malaria incidence has decreased considerably. There is an ethical concern in administering primaquine as a gametocytocide as it does not confer any direct benefit to the treated patient. However, the single low-dose primaquine is most likely associated with very low risk for adverse hematological effects, and WHO recommends its use even without prior G6PD testing. In our opinion, clinical studies including G6PD test should be conducted to assess the safety of low-dose primaquine in African patients. Second, primaquine is effective and necessary for radical treatment of P. vivax and P. ovale, but the standard 14-day treatment (0.25-0.5 mg base/kg/day) is not recommended in patients with G6PD deficiency. Prior G6PD testing is required before prescribing primaquine for radical treatment. The use of primaquine for radical treatment in patients without contraindications does not raise any major ethical problem since the probability of relapse in patients who do not receive anti-hypnozoite treatment can be relatively high and each relapse can cause or aggravate anemia, especially in children. In our opinion, patients with mild or moderate G6PD deficiency should not be treated with primaquine at present. Further clinical studies are necessary to define the role of this drug for radical treatment in G6PD-deficient African patients. Without primaquine, the eventual elimination of P. vivax and P. ovale malaria appears to be very difficult. Updated epidemiological data on G6PD, Duffy antigen, and the current distribution of and burden due to P. vivax and P. ovale are required for a rational use of primaquine in the African continent. Moreover, clinical studies on primaquine are required in Africa.

Malaria control strategies in French armed forces.

Each year, 40,000 French soldiers deploy or travel through malaria-endemic areas. Despite the effective control measures that were successively implemented, malaria remains a public health concern in French armed forces with several important outbreaks and one lethal case every two years. This article describes the malaria control strategy in French armed forces which is based on three combined strategies: i) Anopheles vector control to prevent infection with the implementation of personal protection against vectors (PPAV) adapted to the field living conditions of the troops. ii) Chemoprophylaxis (CP) to prevent the disease based on prescription of effective and well tolerated doxycycline. iii) Management of cases through early diagnosis and appropriate treatment to prevent death. In isolated conditions in endemic areas, rapid diagnosis tests (RDT) are used as first-line tests by military doctors. Treatment of uncomplicated Plasmodium falciparum (P. falciparum) malaria is based either on the piperaquine tetraphosphate-dihydroartemisinin association since 2013, or on the atovaquone-proguanil association. First-line treatment of severe P. falciparum malaria is based on IV artesunate. These measures are associated with constant education of the military, epidemiological surveillance of malaria cases and monitoring of parasite chemosensitivity.

[Delayed diagnosis of Plasmodium falciparum in a soldier in Uganda: false-positive rapid diagnostic test associated with reduced repeats in pfhrp2]. / Retard de diagnostic d'un accès palustre à Plasmodium falciparum chez un militaire en Ouganda : négativité des tests de diagnostic rapide associée à un polymorphisme de l'antigène HRP2.

Rapid diagnostic tests (RDTs) are the best alternative for malaria diagnosis where a microscopic examination cannot be performed. We report here the first case of P. falciparum (false-negative) misdiagnosis in a soldier stationed in Uganda, associated with a reduced number of repeats in the P. falciparum histidine-rich protein 2 gene (pfhrp2). This gene was subsequently sequenced to determine the reason for the discordance between the RDT results and the later microscopic examination. Ten repeats of the type 2 motif AHHAHHAAD and four repeats of the type 7 motif AHHAAD were found. This isolate belongs to the group of non-sensitive parasites (<43 repeats) that are not detected by HRP2 RDTs. This inappropriate case management could have been fatal for the patient. This case confirms the problem of negative RDT results in isolated situations and of basing a therapeutic strategy on these negative results. Investigations should be conducted in Uganda and other areas of Africa to determine the presence and the geographical spread of parasites with pfhrp2 gene deletion to ensure the best performance of RDTs.

Absence of association between ex vivo susceptibility to doxycycline and pftetQ and pfmdt copy numbers in Plasmodium falciparum isolates from Dakar, Senegal.

The objective of this study was to validate the use of pftetQ and pfmdt genes as molecular markers of decreased in vitro susceptibility to doxycycline in 113 Plasmodium falciparum isolates from Dakar, Senegal. The results show that copy numbers of pftetQ and pfmdt, estimated by TaqMan real-time PCR, are not significantly associated with reduced susceptibility to doxycycline in vitro; however, the number of samples with a high doxycycline IC(50) was likely to be too low to derive statistically significant results. Thus, no definitive conclusions could be drawn. The markers should be further tested by analysing more isolates.

Sialome individuality between Aedes aegypti colonies.

Aedes aegypti is responsible for the transmission of arboviruses. The Yellow Fever, Dengue and Chikungunya viruses are transmitted to the vertebrate host by injection of infected saliva during the blood meal of its vectors. Saliva contains different components with various biochemical activities; anti-hemostatic, angiogenic, inflammatory, and immunomodulatory. This work compares the sialomes of three Ae. aegypti colonies (Rockefeller, PAEA, and Formosus), where the repertoire of salivary proteins from these colonies was analyzed by a proteomic approach. This study indicated that major proteins were detectable in the three colonies. However, differences in the abundance of some saliva proteins have been observed between the three Ae. aegypti colonies.

[Molecular markers of Plasmodium falciparum drug resistance]. / Marqueurs moléculaires des résistances de Plasmodium falciparum aux antipaludiques.

The main Plasmodium falciparum genes known to be associated with drug resistance are pfcrt, pfmdr1, pfdhfr pfdhps, pfcytb, pfmrp, pfnhe1, pfmdt, pfserca and pftetQ. Molecular markers for resistance to chloroquine, amodiaquine, mefloquine, sulfadoxine-pyrimethamine, cycloguanil and atovaquone have been validated and used to predict the parasitological and clinical efficacy of these drugs (i.e. based on in vivo tests). These molecular markers have numerous advantages. They allow evaluation of large series of samples in less time than in vivo tests. Collection, transport and storage of samples are much easier than for in vitro tests. These markers can be used for epidemiological monitoring of resistance for an entire country as well as for prediction of therapeutic outcome for a single individual. Development of high-throughput molecular biology techniques, availability of the nucleotidic sequences of P. falciparum genomes, and close collaboration between fundamental researcher workers, clinical practitioners, and officials in charge of the national malaria control programs are major assets in the research and development of molecular markers for P. falciparum resistance to antimalarial drugs.

[Distinguishing Plasmodium falciparum populations for clinical research]. / Distinguer les populations de Plasmodium falciparum en recherche clinique.

The genetic diversity of Plasmodium falciparum is generally high. Study of this diversity is useful to differentiate the strains present in an individual before and after malaria treatment or to check if several individuals have been infected by the same parasites. Interpretation of the in vivo test recommended by the WHO to evaluate antimalarial drug efficacy requires distinguishing recrudescence from new infection when recurrent parasitemia suggests the possibility of therapeutic failure and antimalaria resistance. For this purpose, molecular biology techniques such as nested-PCRs are becoming increasingly available and can now be used in most endemic areas. An effort has been made to standardize P. falciparum genotyping carried out to distinguish recrudescence from new infection. Standardization has focused not only on genotyping methods but also on interpretation criteria. Compliance with these recommendations should improve the quality of clinical research and allow comparison of data from different centers.

[New use of primaquine for malaria]. / Le nouvel âge de la primaquine contre le paludisme.

Primaquine is the only available drug to treat Plasmodium vivax liver stages (hypnozoites). It has been used for more than five decades and is now included in an increasing number of clinical guidelines. The major concern is induced hemolysis when administered to glucose-6-phosphate-dehydrogenase deficient patients. Primaquine could be used for causal prophylaxis during and after exposure or for presumptive antirelapse therapy (PART) in case of high exposure to P. vivax. A radical cure is used to avoid relapse for patients with a confirmed bloodstream infection with P. vivax or P. ovale. In France, primaquine is not approved for prevention and treatment and its use requires a specific temporary authorization.

[Proteomic analysis and parasitosis: principles and applications]. / Analyses protéomiques et parasitoses: principes et applications.

O'Farrel described a method allowing two-dimensional (2D) protein separation more than 30 years ago. Since then the original technique has made enormous progress. This progress has been accompanied by advances in mass spectrometry technology as well as various genome-sequencing programs. Today 2D electrophoresis has become the workhorse of proteomics, allowing resolution of complex structures containing thousands of proteins and providing a global view of the state of a proteome. This article presents the different steps and limitations of proteomic analysis: preparation of biological material, 2D electrophoresis, protein detection systems, and available tools for protein identification. Alternative proteomic approaches to 2D electrophoresis are also presented. A few applications are described as examples to illustrate the utility of proteomic analysis for studying the mechanisms underlying virulence, resistance to antimalarial therapies and immune response against pathologic agents.

[Cyclines and malaria]. / Cyclines et paludisme.

The emergence and rapid spread of Plasmodium falciparum resistance to various antimalarials compounds is gradually reducing the clinician's options for treating malaria and for adapting prophylaxis to each traveler and destination. In this context doxycycline is an increasingly useful alternative except in individuals with contraindications, mainly children under the age of eight and pregnant women. Already used successfully in association with quinine for treatment of malaria in areas with multiresistance, doxycline has also proven to be effective and well tolerated for prophylaxis of malaria. No resistance to doxycycline has been observed to date. Most reported prophylactic failures have been related to poor compliance during the month following return from the endemic zone. The mechanisms of action of doxycycline on the parasite are still unclear. Identification of the molecular targets of doxycycline would open the way for the design of more active structural analogues with longer half-life.

First record of Aedes albopictus in Gabon.

Aedes albopictus were collected in the French military camp of Libreville, Estuaire Province, Gabon, from January to March 2007 by human landing catches during an entomological evaluation of malaria transmission. Inspection of potential larval habitats within and outside the camp showed that Ae. albopictus was found only in artificial containers (discarded tires and small water containers). Associated species of mosquito larvae were Ae. aegypti (L.) and Culex quinquefasciatus. At the same time, Ae. albopictus adults and larvae were also collected from discarded tires in Tcheungue near Port Gentil, Ogoue Maritime Province. Ae. albopictus seems to be established in this part of Gabon's littoral. Further studies are necessary to investigate the extension of Ae. albopictus establishment throughout the country.

[Malaria vaccines: prospects and reality]. / Vaccins contre le paludisme: perspectives et réalité

The development of a malaria vaccine has been accelerating in the last ten years. The number of clinical trials has increased and some malaria antigens have been tested in endemic areas. No potential vaccine has yet shown sufficient and lasting efficacy to justify its inclusion in a public health program. However, trials have unambiguously shown that some level of anti-malaria clinical immunity can be achieved by vaccination, both in experimental and in field conditions. Advances in malaria vaccine development are presented.

In vitro activity of iron-binding compounds against Senegalese isolates of Plasmodium falciparum.

OBJECTIVES: The in vitro activities of FR160, a synthetic catecholate siderophore, and two iron-binding agents, desferrioxamine and doxycycline, were evaluated against Plasmodium falciparum isolates. Correlations between these compounds and standard antimalarial drugs (chloroquine, quinine, amodiaquine, pyronaridine, artemether, artesunate, atovaquone, cycloguanil and pyrimethamine) were assessed to determine any degree of cross-resistance. METHODS: Between October 1997 and February 1998, and September and November 1998, 189 P. falciparum isolates were obtained in Dielmo and Ndiop (Dakar). Their susceptibilities were assessed using an isotopic, microwell format, drug susceptibility test. RESULTS: The 137 inhibitory concentrations (IC(50)) values of FR160 ranged from 0.1 to 10 microM and the geometric mean IC(50) was 1.48 microM (95% CI = 1.29-1.68 microM). The geometric mean IC(50) of doxycycline for 121 isolates was 18.9 microM (95% CI = 16.8-21.3 microM) and that of desferrioxamine for 73 isolates was 20.7 microM (95% CI = 17.3-24.8 microM). FR160 was significantly less active against the chloroquine-resistant isolates (P < 0.0001). The mean IC(50)s of doxycycline were significantly higher for the chloroquine-susceptible isolates than for the resistant parasites (P = 0.0447). There was a weak correlation between the responses to FR160, desferrioxamine or doxycycline and those to the other antimalarial compounds (r(2) < 0.22). CONCLUSIONS: The activities of FR160 and desferrioxamine, determined for P. falciparum clones, were confirmed against 137 isolates. The coefficients of determination between the responses to FR160, doxycycline or desferrioxamine and those to all the antimalarial drugs tested are too weak to suggest cross-resistance. FR160 could be a rationale partner to use in combination with doxycycline.