Use of Plasmodium falciparum culture-adapted field isolates for in vitro exflagellation-blocking assay

International audienceA major requirement for malaria elimination is the development of transmission-blocking interventions. In vitro transmission-blocking bioassays currently mostly rely on the use of very few Plasmodium falciparum reference laboratory strains isolated decades ago. To fill a piece of the gap between laboratory experimental models and natural systems, the purpose of this work was to determine if culture-adapted field isolates of P. falciparum are suitable for in vitro transmission-blocking bioassays targeting functional maturity of male gametocytes: exflagellation. Plasmodium falciparum isolates were adapted to in vitro culture before being used for in vitro gametocyte production. Maturation was assessed by microscopic observation of gametocyte morphology over time of culture and the functional viability of male gametocytes was assessed by microscopic counting of exflagellating gametocytes. Suitability for in vitro exflagellation-blocking bioassays was determined using dihydroartemisinin and methylene blue. In vitro gametocyte production was achieved using two isolates from French Guiana and two isolates from Cambodia. Functional maturity of male gametocytes was assessed by exflagellation observations and all four isolates could be used in exflagellation-blocking bioassays with adequate response to methylene blue and dihydroartemisinin. This work shows that in vitro culture-adapted P. falciparum field isolates of different genetic background, from South America and Southeast Asia, can successfully be used for bioassays targeting the male gametocyte to gamete transition, exflagellation

Novel Clinical and Diagnostic Aspects of Antineutrophil Cytoplasmic Antibodies

Antineutrophil cytoplasmic antibodies (ANCA) are the serological hallmark of some idiopathic systemic vasculitides. Besides the investigation of ANCA-associated vasculitis (AAV) and constant effort for a standardized nomenclature and classification of the AAV, amain focus of research during the last few years has been to constantly improve the performance of enzyme immunoassays. With the latest so called third generation ELISA, this goal seemed to be fulfilled. The International Consensus Statement on Testing and Reporting of ANCA gave recommendations for standardized strategies for the serological diagnosis of ANCA. New developments now target the system immanent drawbacks of the respective diagnosticmethods, be it the need for batching and the long time to result for ELISA, or the high likelihood of error and subjectivity of indirect immunofluorescence (IIF). Randomaccess technology andmultiplexing for solid phase assays aswell as digital imaging for IIF are toolswhichmay help to expedite and simplify routine diagnostics in the lab and in emergency settings. Recent findings indicate that PR3-ANCA have clinical utility beyond the diagnosis of AAV. PR3-ANCA can also serve as an aid for the differentiation between ulcerative colitis (UC) and Crohn’s disease (CrD) and the stratification of UC patients. This review provides a detailed review of what is known about ANCA and highlights the latest research and state-of-the-art developments in this area

Solar Jet Hunter: a citizen science initiative to identify coronal jets in EUV data sets

Context. Solar coronal jets seen in EUV are ubiquitous on the Sun, have been found in and at the edges of active regions, at the boundaries of coronal holes, and in the quiet Sun. Jets have various shapes, sizes, brightness, velocities and duration in time, which complicates their detection by automated algorithms. So far, solar jets reported in the Heliophysics Event Knowledgebase (HEK) have been mostly reported by humans looking for them in the data, with different levels of precision regarding their timing and positions. Aims. We create a catalogue of solar jets observed in EUV at 304 {\AA} containing precise and consistent information on the jet timing, position and extent. Methods. We designed a citizen science project, "Solar Jet Hunter", on the Zooniverse platform, to analyze EUV observations at 304 {\AA} from the Solar Dynamic Observatory/Atmospheric Imaging Assembly (SDO/AIA). We created movie strips for regions of the Sun in which jets have been reported in HEK and ask the volunteers to 1) confirm the presence of at least one jet in the data and 2) report the timing, position and extent of the jet. Results. We report here the design of the project and the results obtained after the analysis of data from 2011 to 2016. 365 "coronal jet" events from HEK served as input for the citizen science project, equivalent to more than 120,000 images distributed into 9,689 "movie strips". Classification by the citizen scientists resulted with only 21% of the data containing a jet, and 883 individual jets being identified. Conclusions. We demonstrate how citizen science can enhance the analysis of solar data with the example of Solar Jet Hunter. The catalogue of jets thus created is publicly available and will enable statistical studies of jets and related phenomena. This catalogue will also be used as a training set for machines to learn to recognize jets in further data sets

Connecting solar flare hard X-ray spectra to in situ electron spectra A comparison of RHESSI and STEREO/SEPT observations

Aims. We aim to constrain the acceleration, injection, and transport processes of flare-accelerated energetic electrons by comparing their characteristics at the Sun with those injected into interplanetary space.Methods. We have identified 17 energetic electron events well-observed with the SEPT instrument aboard STEREO which show a clear association with a hard X-ray (HXR) flare observed with the RHESSI spacecraft. We compare the spectral indices of the RHESSI HXR spectra with those of the interplanetary electrons. Because of the frequent double-power-law shape of the in situ electron spectra, we paid special attention to the choice of the spectral index used for comparison.Results. The time difference between the electron onsets and the associated type III and microwave bursts suggests that the electron events are detected at 1 AU with apparent delays ranging from 9 to 41 min. While the parent solar activity is clearly impulsive, also showing a high correlation with extreme ultraviolet jets, most of the studied events occur in temporal coincidence with coronal mass ejections (CMEs). In spite of the observed onset delays and presence of CMEs in the low corona, we find a significant correlation of about 0.8 between the spectral indices of the HXR flare and the in situ electrons. The correlations increase if only events with significant anisotropy are considered. This suggests that transport effects can alter the injected spectra leading to a strongly reduced imprint of the flare acceleration.Conclusions. We conclude that interplanetary transport effects must be taken into account when inferring the initial acceleration of solar energetic electron events. Although our results suggest a clear imprint of flare acceleration for the analyzed event sample, a secondary acceleration might be present which could account for the observed delays. However, the limited and variable pitch-angle coverage of SEPT could also be the reason for the observed delays

Exploring impulsive solar magnetic energy release and particle acceleration with focused hard X-ray imaging spectroscopy

How impulsive magnetic energy release leads to solar eruptions and how those eruptions are energized and evolve are vital unsolved problems in Heliophysics. The standard model for solar eruptions summarizes our current understanding of these events. Magnetic energy in the corona is released through drastic restructuring of the magnetic field via reconnection. Electrons and ions are then accelerated by poorly understood processes. Theories include contracting loops, merging magnetic islands, stochastic acceleration, and turbulence at shocks, among others. Although this basic model is well established, the fundamental physics is poorly understood. HXR observations using grazing-incidence focusing optics can now probe all of the key regions of the standard model. These include two above-the-looptop (ALT) sources which bookend the reconnection region and are likely the sites of particle acceleration and direct heating. The science achievable by a direct HXR imaging instrument can be summarized by the following science questions and objectives which are some of the most outstanding issues in solar physics (1) How are particles accelerated at the Sun? (1a) Where are electrons accelerated and on what time scales? (1b) What fraction of electrons is accelerated out of the ambient medium? (2) How does magnetic energy release on the Sun lead to flares and eruptions? A Focusing Optics X-ray Solar Imager (FOXSI) instrument, which can be built now using proven technology and at modest cost, would enable revolutionary advancements in our understanding of impulsive magnetic energy release and particle acceleration, a process which is known to occur at the Sun but also throughout the Universe

Land cover, land use and malaria in the Amazon: a systematic literature review of studies using remotely sensed data

The nine countries sharing the Amazon forest accounted for 89% of all malaria cases reported in the Americas in 2008. Remote sensing can help identify the environmental determinants of malaria transmission and their temporo-spatial evolution. Seventeen studies characterizing land cover or land use features, and relating them to malaria in the Amazon subregion, were identified. These were reviewed in order to improve the understanding of the land cover/use class roles in malaria transmission. The indicators affecting the transmission risk were summarized in terms of temporal components, landscape fragmentation and anthropic pressure. This review helps to define a framework for future studies aiming to characterize and monitor malaria

COIL: a methodology for evaluating malarial complexity of infection using likelihood from single nucleotide polymorphism data.

International audienceComplex malaria infections are defined as those containing more than one genetically distinct lineage of Plasmodium parasite. Complexity of infection (COI) is a useful parameter to estimate from patient blood samples because it is associated with clinical outcome, epidemiology and disease transmission rate. This manuscript describes a method for estimating COI using likelihood, called COIL, from a panel of bi-allelic genotyping assays. COIL assumes that distinct parasite lineages in complex infections are unrelated and that genotyped loci do not exhibit significant linkage disequilibrium. Using the population minor allele frequency (MAF) of the genotyped loci, COIL uses the binomial distribution to estimate the likelihood of a COI level given the prevalence of observed monomorphic or polymorphic genotypes within each sample. COIL reliably estimates COI up to a level of three or five with at least 24 or 96 unlinked genotyped loci, respectively, as determined by in silico simulation and empirical validation. Evaluation of COI levels greater than five in patient samples may require a very large collection of genotype data, making sequencing a more cost-effective approach for evaluating COI under conditions when disease transmission is extremely high. Performance of the method is positively correlated with the MAF of the genotyped loci. COI estimates from existing SNP genotype datasets create a more detailed portrait of disease than analyses based simply on the number of polymorphic genotypes observed within samples. The capacity to reliably estimate COI from a genome-wide panel of SNP genotypes provides a potentially more accurate alternative to methods relying on PCR amplification of a small number of loci for estimating COI. This approach will also increase the number of applications of SNP genotype data, providing additional motivation to employ SNP barcodes for studies of disease epidemiology or control measure efficacy. The COIL program is available for download from GitHub, and users may also upload their SNP genotype data to a web interface for simple and efficient determination of sample COI

Plasmodium chabaudi chabaudi malaria parasites can develop stable resistance to atovaquone with a mutation in the cytochrome b gene

<p>Abstract</p> <p>Background</p> <p><it>Plasmodium falciparum</it>, has developed resistance to many of the drugs in use. The recommended treatment policy is now to use drug combinations. The atovaquone-proguanil (AP) drug combination, is one of the treatment and prophylaxis options. Atovaquone (ATQ) exerts its action by inhibiting plasmodial mitochondria electron transport at the level of the cytochrome bc1 complex. <it>Plasmodium falciparum in vitro </it>resistance to ATQ has been associated with specific point mutations in the region spanning codons 271-284 of the <it>cytochrome b </it>gene. ATQ -resistant <it>Plasmodium yoelii </it>and <it>Plasmodium berghei </it>lines have been obtained and resistant lines have amino acid mutations in their CYT <it>b </it>protein sequences. <it>Plasmodium chabaudi </it>model for studying drug-responses and drug-resistance selection is a very useful rodent malaria model but no ATQ resistant parasites have been reported so far. The aim of this study was to determine the ATQ sensitivity of the <it>P. chabaudi </it>clones, to select a resistant parasite line and to perform genotypic characterization of the <it>cytb </it>gene of these clones.</p> <p>Methods</p> <p>To select for ATQ resistance, <it>Plasmodium. chabaudi chabaudi </it>clones were exposed to gradually increasing concentrations of ATQ during several consecutive passages in mice. <it>Plasmodium chabaudi cytb </it>gene was amplified and sequenced.</p> <p>Results</p> <p>ATQ resistance was selected from the clone AS-3CQ. In order to confirm whether an heritable genetic mutation underlies the response of AS-ATQ to ATQ, the stability of the drug resistance phenotype in this clone was evaluated by measuring drug responses after (i) multiple blood passages in the absence of the drug, (ii) freeze/thawing of parasites in liquid nitrogen and (iii) transmission through a mosquito host, <it>Anopheles stephensi</it>. ATQ resistance phenotype of the drug-selected parasite clone kept unaltered. Therefore, ATQ resistance in clone AS-ATQ is genetically encoded. The Minimum Curative Dose of AS-ATQ showed a six-fold increase in MCD to ATQ relative to AS-3CQ.</p> <p>Conclusions</p> <p>A mutation was found on the <it>P. chabaudi cytb </it>gene from the AS-ATQ sample a substitution at the residue Tyr268 for an Asn, this mutation is homologous to the one found in <it>P. falciparum </it>isolates resistant to ATQ.</p

Atmospheric neutrinos with three flavor mixing

We analyze the atmospheric neutrino data in the context of three flavor neutrino oscillations taking account of the matter effects in the earth. With the hierarchy among the vacuum mass eigenvalues $\mu_3^2 \gg \mu_2^2 \geq \mu_1^2$, the solution of the atmospheric neutrino problem depends on $\delta_{31}=\mu_3^2 - \mu_1^2$ and the $13$ and $23$ mixing angles $\phi$ and $\psi$. Whereas the sub-GeV atmospheric neutrino data imposes only a lower limit on $\delta_{31} > 10^{-3} eV^2$, the zenith angle dependent suppression observed in the multi-GeV data limits $\delta_{31}$ from above also. The allowed regions of the parameter space are strongly constrained by the multi-GeV data. Combined with our earlier solution to the solar neutrino problem which depends on $\delta_{21}= \mu_2^2-\mu_1^2$ and the $12$ and $13$ mixing angles $\omega$ and $\phi$, we have obtained the ranges of values of the five neutrino parameters which solve both the solar and the atmospheric neutrino problems simultaneously.Comment: 21 pages Revtex, 7 figures as 7 ps files, Zenith angle binned multi-Gev analysis redone with the Kamiokande detector efficiencies include