Judith Fillaux

Antimalarial Activity of Simalikalactone E, a New Quassinoid from Quassia amara L. (Simaroubaceae)

ABSTRACT We report the isolation and identification of a new quassinoid named simalikalactone E (SkE), extracted from a widely used Amazonian antimalarial remedy made out of Quassia amara L. (Simaroubaceae) leaves. This new molecule inhibited the growth of Plasmodium falciparum cultured in vitro by 50%, in the concentration range from 24 to 68 nM, independently of the strain sensitivity to chloroquine. We also showed that this compound was able to decrease gametocytemia with a 50% inhibitory concentration sevenfold lower than that of primaquine. SkE was found to be less toxic than simalikalactone D (SkD), another antimalarial quassinoid from Q. amara, and its cytotoxicity on mammalian cells was dependent on the cell line, displaying a good selectivity index when tested on nontumorogenic cells. In vivo, SkE inhibited murine malaria growth of Plasmodium vinckei petteri by 50% at 1 and 0.5 mg/kg of body weight/day, by the oral or intraperitoneal routes, respectively. The contribution of quassinoids as a source of antimalarial molecules needs therefore to be reconsidered.

Schistosomiasis Haematobium, Corsica, France

To the Editor: In Europe, urinary schistosomiasis (1) has previously been detected only in Portugal, where this focus disappeared during the 1950s (2). However, freshwater snails of the species Bulinus contortus, B. truncatus, and Planorbarius metidjensis, which are recognized intermediate hosts for Schistosoma haematobium trematodes, have been found in Portugal (3), Spain (4), and Corsica (5,6). This finding suggested that autochthonous schistosomiasis could re-emerge in southern Europe if these mollusks become infected. We report a probable focus for transmission of schistosomiasis haematobium in Corsica, France. In March 2014, a 4-year-old girl (index case-patient) from France was referred to the Toulouse University Hospital (Toulouse, France), with gross hematuria. Ultrasonography and cystoscopic examination of the bladder detected a polyp. Examination of the polyp for parasites identified bodies that were consistent with schistosome eggs. Parasitologic examination of urine confirmed schistosomiasis by detecting viable S. haematobium eggs. The parents of the girl (family A) did not report any stay or travel in an area to which urinary schistosomiasis was endemic; they reported summer holidays only in Mallorca in the Balearic Islands (Spain) and Corsica. However, her father reported that since 2012, he had experienced gross hematuria that had been evaluated by standard urologic investigations but not by cystoscopy; no etiology was determined. Parasitologic urinalysis in our hospital department showed numerous viable S. haematobium eggs in the father’s urine. The parents of the index case-patient also reported that an 8-year-old boy in a friend’s family (family B), who shared summer vacations with them had exhibited gross hematuria since February 2013. A third family (family C) was also investigated because they also spent holidays in Corsica with families A and B. Families B and C had also spent a summer in Mallorca, but they denied any contact with freshwater. Of 11 French native-born members of the 3 families, 6 had positive results for S. haematobium by urine examination. All case-patients had specific positive immunodiagnostic results by an ELISA that used S. mansoni extracts and by indirect hemagglutination. In addition, 2 family members who had a negative result by urine examination had a positive serologic result. Spending summer vacations in the same village in Corsica (Sainte-Lucie de Porto-Vecchio), where members of the 3 families had bathed at least once per holiday period in the Cavu River, was the epidemiologically prominent feature that linked these persons. Families A and C were in Sainte-Lucie de Porto-Vecchio in August 2011, and families A, B, and C were in the same location in August 2013. During these investigations, we were contacted by the Department of Tropical Medicine, Dusseldorf University Hospital (Dusseldorf, Germany), because a 10-year-old boy and his father had been given diagnoses of schistosomiasis haematobium on the basis of positive urinalysis results for S. haematobium eggs. Two other members of this family (5 persons) had a positive immunodiagnostic result. Locations of previous vacations for this family outside Germany included Spain (not the Balearic Islands) and Corsica, where they bathed frequently in the Cavu River. These epidemiologic findings provide strong circumstantial evidence supporting the presence of a previously unrecognized focus of urinary schistosomiasis in Corsica. We performed molecular analysis of schistosomal miracidia DNA. The second internal transcribed spacer region of the ribosomal gene complex (7,8) was amplified and sequenced. Viable eggs obtained from the patients in France were those of S. haematobium. Additional molecular investigations are being conducted to assess genetic diversity of this isolate from Corsica and the geographic origin of the introduced parasite. The malacologic situation in Sainte Lucie de Porto-Vecchio was investigated during May 12–19, 2014; three rivers (Figure) were included in the survey. Four sites were sampled in the Cavu River, and B. truncatus snails were found in 3 sites that corresponded to bathing areas (site 1: 41°43′53.57″N, 9°17′36.70″E; site 2: 41°43′22.13″N, 9°17′59.87″E; site 3: 41°42′8.40″N, 9°21′5.82″E). Snails were also found in the nearby Tarcu River (site 5) and Osu River (site 6). These findings confirmed previous data for the presence of B. truncatus snails in Corsica (5,6). Water temperature was recorded at 11:00 am at these 3 sites (range 15°C–16°C). This temperature range is not optimal for the snail intermediate host stage of the parasite life cycle (9,10). Of 148 live snails that were obtained in the Cavu River, none were infected with schistosome cercariae. Figure Corsica, France, showing malacologic survey sampling sites (oval) in 3 rivers (Tarcu, Cavu, and Osu). Bulinus truncatus snails were found at sites 1, 2, 3, 5, and 6. Data from the field survey and epidemiologic information for the cases in France and Germany, indicated transmission of schistosomiasis haematobium in the Cavu River in southeastern Corsica in 2011 and 2013. Additional supportive evidence is the fact that the father of the index case-patient had gross hematuria in 2012 and 2013. Two hypotheses are proposed to account for this situation. The first hypothesis is that the parasite (i.e., schistosome eggs) was transmitted by an infected person into the Cavu River in June or July 2011, when environmental conditions were favorable for snail infection. However, questions arise about survival of infected snails during the winter and their ability to reinfect the area during the following summers in 2012 and 2013. The second hypothesis is that schistosome eggs were spread by infected persons at the beginning of summer and caused a permanent transmission cycle in this focus. This situation would be difficult to control. Additional information should be obtained by a long-term malacologic survey to detect infected mollusks in this region.

Assessment of Aspergillus sensitization or persistent carriage as a factor in lung function impairment in cystic fibrosis patients

Background: Cystic fibrosis (CF) patients presenting with persistent carriage of, or sensitization to, Aspergillus fumigatus are often treated with antifungal therapies because the presence of the fungus is commonly thought to impede lung function, even in the absence of allergic bronchopulmonary aspergillosis (ABPA). The aim of this study was to assess Aspergillus-related status modulating the forced expiratory volume in 1 s (FEV1) of CF patients. Methods: From 1995 to 2007, 251 patients were evaluated. Demographic data, cystic fibrosis transmembrane conductance regulator gene (CFTR) mutations, body mass index, and FEV1 were recorded. The presence of A. fumigatus and Pseudomonas aeruginosa in sputum and the levels of A. fumigatus precipitin, total IgE (t-IgE), and specific anti-A. fumigatus IgE (Af-IgE) were determined. Patients were divided into 3 groups: (1) ABPA: A. fumigatus precipitin ≥3 lines, Af-IgE > 0.35 IU/ml, and t-IgE ≥500 IU/ml; (2) sensitization: Af-IgE > 0.35 IU/ml but t-IgE < 500 IU/ml; and (3) persistent carriage: Af-IgE ≤ 0.35 IU/ml with either an A. fumigatus persistent positive culture or an A. fumigatus precipitin ≥3 lines, provided this serological finding had been found associated with at least 1 A. fumigatus-positive culture. The remaining patients represented the control group. A multivariate analysis was carried out with FEV1 as the outcome variable. Results: ABPA, sensitization, and persistent carriage were significantly associated with a larger decline in FEV1 compared with the control group, with odds ratios of 15.9, 14.9, and 10.7, respectively. This association was independent of other associated factors (P. aeruginosa transient detection, age, being underweight, and low FEV1 at baseline). Conclusions: In addition to ABPA, sensitization and persistent carriage appear to have an impact on pulmonary function in CF patients.

Imported Plasmodium knowlesi Malaria in a French Tourist Returning from Thailand

We report a case of imported Plasmodium knowlesi malaria in a French tourist following a vacation in Thailand. This case shows, first, tourists may contract knowlesi malaria even only staying on the beach and second, the diagnosis remains difficult, even with polymerase chain reaction methods.

Toxoplasmic encephalitis IRIS in HIV-infected patients: a case series and review of the literature

Background Toxoplasmic encephalitis associated with immune reconstitution inflammatory syndrome (TE-IRIS) is rarely described. Methods To identify TE-IRIS cases, the authors performed a retrospective study of all HIV-infected patients diagnosed as having TE in our unit between January 2000 and June 2009, and a review of published cases. Results Three patients out of 65 toxoplasmic encephalitis (TE) cases, together with six from the literature, fulfilled the unmasking TE-IRIS definition. None fulfilled the paradoxical TE-IRIS definition. TE occurred within a median time of 48.5 days (IQ25–75 21–56) after starting antiretroviral therapy. Cases did not have distinctive clinical or neuroimaging features from TE occurring without immune reconstitution. However: (1) cases occurred at a median CD4 lymphocytes count of 222/μl (IQ25–75 160–280); (2) TE occurred in five patients who were supposed to take an effective chemoprophylaxis; (3) two patients had a brain biopsy showing an intense angiocentric inflammatory infiltrating with predominantly CD8 lymphocytes; (4) in one patient, the abnormal length of evolution under treatment might be due to the heightened immune response. Conclusion Although rare, unmasking TE-IRIS exists and might occur despite effective prophylaxis and an unusually high CD4 lymphocyte count. Immune reconstitution inflammatory syndrome does not modify TE diagnosis and treatment but might extend its clinical course.

Evidence for a permanent presence of schistosomiasis in Corsica, France, 2015

We present a case of acute schistosomiasis acquired in Corsica after bathing in the Cavu River during the summer of 2015. The diagnosis was made following epidemiological, laboratory and serological assessments. After a previous outbreak of urogenital schistosomiasis during the summer of 2013, when more than 120 infections were diagnosed, this further case indicates transmission was still effective in 2015, thus suggesting a permanent presence of schistosomiasis in Corsica.

Aspergillus sensitization or carriage in cystic fibrosis patients.

Background: Aspergillus fumigatus (Af) sensitization and persistent carriage are deleterious to lung function, but no consensus has been reached defining these medical entities. This work aimed to identify possible predictive factors for patients who become sensitized to Af, compared with a control group of non-sensitized Af carriers. Methods: Between 1995 and 2007, 117 pediatric patients were evaluated. Demographic data, CFTR gene mutations, body mass index and FEV1 were recorded. The presence of Af in sputum, the levels of Af-precipitin, total IgE (t-IgE) and specific IgE to Af (Af-IgE) were determined. Patients were divided into 2 groups: (1) “sensitization”: level of Af-IgE > 0.35 IU/mL with t-IgE level < 500 IU/mL and (2) “persistent or transient carriage”: Af-IgE level ⩽ 0.35 IU/mL with either an Af transient or persistent positive culture. A survival analysis was performed with the appearance of Af-IgE in serum as an outcome variable. Results: Severe mutation (hazard ratio = 3.2), FEV1 baseline over 70% of theoretical value (hazard ratio = 4.9), absence of Pa colonization, catalase activity and previous azithromycin administration (hazard ratio = 9.8, 4.1 and 1.9, respectively) were predictive factors for sensitization. We propose a timeline of the biological events and a tree diagram for risk calculation. Conclusions: Two profiles of cystic fibrosis patients can be envisaged: (1) patients with nonsevere mutation but low FEV1 baselines are becoming colonized with Af or (2) patients with high FEV1 baselines who present with severe mutation are more susceptible to the Af sensitization and then to the presentation of an allergic bronchopulmonary aspergillosis event.

An extraction method of positive blood cultures for direct identification of Candida species by Vitek MS matrix-assisted laser desorption ionization time of flight mass spectrometry

Candida spp. are an important cause of nosocomial bloodstream infections. Currently, complete identification of yeasts with conventional methods takes several days. We report here the first evaluation of an extraction method associated with the Vitek MS matrix-assisted laser desorption ionization time of flight mass spectrometry for direct identification of Candida species from positive blood cultures. We evaluated this protocol with blood cultures that were inoculated with reference and routine isolates (eight reference strains, 30 patients isolates and six mixed cultures containing two strains of different Candida species), or from patients with candidemia (28 isolates). This method performed extremely well (97% correct identification) with blood cultures of single Candida spp. and significantly reduced the time of diagnosis. Nevertheless, subculture remains indispensable to test fungal resistance and to detect mixed infections.

Toxoplasmose et grossesse

Resume La toxoplasmose est une parasitose cosmopolite, tres repandue chez l’homme et l’animal due a Toxoplasma gondii protozoaire intracellulaire. L’infection en cours de la grossesse peut provoquer une infection congenitale dont la manifestation clinique la plus frequente est la chorioretinite. Elle est depistee des la naissance ou plus tard avant 10 ans dans 95 % des cas. L’enquete perinatale effectuee en France en 2003 a montre que la prevalence d’une immunite chez les femmes enceintes etait de 44 %. Il est essentiel pour prevenir l’infection de fournir des recommandations hygieno-dietetiques, lavage des mains et des instruments de cuisine, cuisson des aliments (viande et legumes), peler les fruits, porter des gants pour tout contact avec la terre, ne pas changer la litiere du chat…). La prevention impose aux femmes enceintes seronegatives une surveillance serologique mensuelle. Le nombre de cas de toxoplasmose congenitale a ete estime en 2000 a 700 par an, soit une incidence de 1 pour 1 000 naissances vivantes. Un traitement precoce maternel par la spiramycine puis par l’association pyrimethamine et sulfamides (sulfadiazine ou sulfadoxine) en cas d’infection fœtale prouvee par le diagnostic prenatal ou neonatal reduit le risque de manifestations cliniques.

Real-Time PCR Assay for the Diagnosis of Pneumocystis jirovecii Pneumonia

Pneumocystis jirovecii is a common cause of life-threatening pneumonia among immunocompromised patients. Since P. jirovecii cannot be cultured, specific identification of it depends on examining respiratory specimens. In the last decade, PCR has been developed which allows the detection of very low levels of P. jirovecii not detectable by routine histochemical staining. We have shown that the direct immunofluorescence assay can be replaced by a real-time PCR assay given its feasibility, sensitivity, and specificity, for the detection of P. jirovecii. A negative PCR, performed on a LightCycler System(®), enables a diagnosis of Pneumocystis jirovecii pneumonia (PjP) to be excluded, and the semiquantitative result with the application of some cutoff values can have a role in distinguishing between colonized or subclinically infected patients and PjP patients.