Cryptosporidium Species and Subtypes and Clinical Manifestations in Children, Peru

One-sentence summary for table of contents: Different genotypes and subtypes are linked to different clinical manifestations

A Screening Pipeline for Antiparasitic Agents Targeting Cryptosporidium Inosine Monophosphate Dehydrogenase

Persistent diarrhea is a leading cause of illness and death among impoverished children, and a growing share of this disease burden can be attributed to the parasite Cryptosporidium. There are no vaccines to prevent Cryptosporidium infection, and the treatment options are limited and unreliable. Critically, no effective treatment exists for children or adults suffering from AIDS. Cryptosporidium presents many technical obstacles for drug discovery; perhaps the most important roadblock is the difficulty of monitoring drug action. Here we have developed a set of methods to accelerate the drug discovery process for cryptosporidiosis. We exploit the opportunities for experimental manipulation in the related parasite Toxoplasma to genetically engineer a Cryptosporidium model. This new model parasite mirrors the metabolism of Cryptosporidium for a particularly promising drug target that supplies the building blocks for DNA and RNA. Drug effectiveness can be assayed through simple fluorescence measurements for many candidates. Using this assay as an initial filter, and adapting other assays to a high throughput format, we identify several novel chemical compounds that exhibit markedly improved anti-cryptosporidial activity and excellent selectivity

The Cryptosporidium parvum Kinome

<p>Abstract</p> <p>Background</p> <p>Hundreds of millions of people are infected with cryptosporidiosis annually, with immunocompromised individuals suffering debilitating symptoms and children in socioeconomically challenged regions at risk of repeated infections. There is currently no effective drug available. In order to facilitate the pursuit of anti-cryptosporidiosis targets and compounds, our study spans the classification of the <it>Cryptosporidium parvum </it>kinome and the structural and biochemical characterization of representatives from the CDPK family and a MAP kinase.</p> <p>Results</p> <p>The <it>C</it>. <it>parvum </it>kinome comprises over 70 members, some of which may be promising drug targets. These <it>C. parvum </it>protein kinases include members in the AGC, Atypical, CaMK, CK1, CMGC, and TKL groups; however, almost 35% could only be classified as OPK (other protein kinases). In addition, about 25% of the kinases identified did not have any known orthologues outside of <it>Cryptosporidium spp</it>. Comparison of specific kinases with their <it>Plasmodium falciparum </it>and <it>Toxoplasma gondii </it>orthologues revealed some distinct characteristics within the <it>C. parvum </it>kinome, including potential targets and opportunities for drug design. Structural and biochemical analysis of 4 representatives of the CaMK group and a MAP kinase confirms features that may be exploited in inhibitor design. Indeed, screening <it>Cp</it>CDPK1 against a library of kinase inhibitors yielded a set of the pyrazolopyrimidine derivatives (PP1-derivatives) with IC₅₀values of < 10 nM. The binding of a PP1-derivative is further described by an inhibitor-bound crystal structure of <it>Cp</it>CDPK1. In addition, structural analysis of <it>Cp</it>CDPK4 identified an unprecedented Zn-finger within the CDPK kinase domain that may have implications for its regulation.</p> <p>Conclusions</p> <p>Identification and comparison of the <it>C. parvum </it>protein kinases against other parasitic kinases shows how orthologue- and family-based research can be used to facilitate characterization of promising drug targets and the search for new drugs.</p

Charity trustees' guide

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First mesocosm experiments to study the impacts of ocean acidification on plankton communities in the NW Mediterranean Sea (MedSeA project)

There is a growing international interest in studying the effects of ocean acidification on plankton communities that play a major role in the global carbon cycle and in the consumption of atmospheric CO2 via the so-called biological pump. Recently, several mesocosm experiments reported on the effect of ocean acidification on marine plankton communities, although the majority were performed in eutro- phic conditions or following nutrient addition. The objective of the present study was to perform two mesocosm experiments in the oligo- to meso-trophic Northwestern Mediterranean Sea during two seasons with contrasting environmental conditions: in summer 2012 in the Bay of Calvi (Corsica, France) and in winter 2013 in the Bay of Villefranche (France). This paper describes the objectives of these ex- periments, the study sites, the experimental set-up and the environmental and experimental conditions during the two experiments. The 20-day experiment in the Bay of Calvi was undoubtedly representative of summer conditions in the Northwestern Mediterranean Sea with low nutrient and chlorophyll a concentrations, warm waters and high surface solar irradiance. In contrast, the winter experiment, which was reduced to 12 days because of bad weather conditions, failed to reproduce the mesotrophic con- ditions typical of the wintertime in this area. Indeed, a rapid increase in phytoplankton biomass during the acidification phase led to a strong decrease in nitrate concentrations and an unrealistic N and P co- limitation at this period of the year. An overview of the 11 other papers related to this study and pub- lished in this special issue is provided.MEDiterranean Sea Acidification in a changing climat

Limited impact of ocean acidification on phytoplankton community structure and carbon export in an oligotrophic environment: Results from two short-term mesocosm studies in the Mediterranean Sea

International audienceModifications in the strength of the biological pump as a consequence of ocean acidification, whether positive or negative, have the potential to impact atmospheric CO2 and therefore climate. So far, most plankton community perturbation studies have been performed in nutrient-rich areas although there are some indications that CO2-dependent growth could differ in nutrient-replete vs. -limited regions and with different community compositions. Two in situ mesocosm experiments were performed in the NW Mediterranean Sea during two seasons with contrasted environmental conditions: summer oligotrophic stratified waters in the Bay of Calvi vs. winter mesotrophic well-mixed waters in the Bay of Villefranche. Nine mesocosms were deployed for 20 and 12 d, respectively, and subjected to seven CO2 levels (3 controls, 6 elevated levels). Both phytoplankton assemblages were dominated by pico- and nano-phytoplankton cells. Although haptophyceae and dinoflagellates benefited from short-term CO2 enrichment in summer, their response remained small with no consequences on organic matter export due to strong environmental constraints (nutrient availability). In winter, most of the plankton growth and associated nutrient consumption occurred during the 4-day acidification period (before the experimental phase). During the remaining experimental period, characterized by low nutrient availability, plankton growth was minimal and no clear CO2-dependency was found for any of the tested parameters. While there is a strong confidence on the absence of significant effect of short-term CO2 addition under oligotrophic conditions, more investigations are needed to assess the response of plankton communities in winter when vertical mixing and weather conditions are major factors controlling plankton dynamics

Primary marine aerosol emissions from the Mediterranean Sea during pre-bloom and oligotrophic conditions: correlations to seawater chlorophyll <i>a</i> from a mesocosm study

The effect of ocean acidification and changing water conditions on primary (and secondary) marine aerosol emissions is not well understood on a regional or a global scale. To investigate this effect as well as the indirect effect on aerosol that changing biogeochemical parameters can have, ~ 52 m³ pelagic mesocosms were deployed for several weeks in the Mediterranean Sea during both winter pre-bloom and summer oligotrophic conditions and were subjected to various levels of CO₂ to simulate the conditions foreseen in this region for the coming decades. After seawater sampling, primary bubble-bursting aerosol experiments were performed using a plunging water jet system to test both chemical and physical aerosol parameters (10–400 nm). Comparing results obtained during pre-bloom and oligotrophic conditions, we find the same four log-normal modal diameters (18.5 ± 0.6, 37.5 ± 1.4, 91.5 ± 2.0, 260 ± 3.2 nm) describing the aerosol size distribution during both campaigns, yet pre-bloom conditions significantly increased the number fraction of the second (Aitken) mode, with an amplitude correlated to virus-like particles, heterotrophic prokaryotes, TEPs (transparent exopolymeric particles), chlorophyll <i>a</i> and other pigments. Organic fractions determined from kappa closure calculations for the diameter, <i>D</i>_p ~ 50 nm, were much larger during the pre-bloom period (64 %) than during the oligotrophic period (38 %), and the organic fraction decreased as the particle size increased. Combining data from both campaigns together, strong positive correlations were found between the organic fraction of the aerosol and chlorophyll <i>a</i> concentrations, heterotrophic and autotrophic bacteria abundance, and dissolved organic carbon (DOC) concentrations. As a consequence of the changes in the organic fraction and the size distributions between pre-bloom and oligotrophic periods, we find that the ratio of cloud condensation nuclei (CCN) to condensation nuclei (CN) slightly decreased during the pre-bloom period. The enrichment of the seawater samples with microlayer samples did not have any effect on the size distribution, organic content or the CCN activity of the generated primary aerosol. Partial pressure of CO₂, <i>p</i>CO₂, perturbations had little effect on the physical or chemical parameters of the aerosol emissions, with larger effects observed due to the differences between a pre-bloom and oligotrophic environment