Apicomplexan Parasites Co-Opt Host Calpains to Facilitate Their Escape from Infected Cells

Apicomplexan parasites, including Plasmodium falciparum and Toxoplasma gondii (the causative agents of malaria and toxoplasmosis, respectively), are responsible for considerable morbidity and mortality worldwide. These pathogenic protozoa replicate within an intracellular vacuole inside of infected host cells, from which they must escape to initiate a new lytic cycle. By integrating cell biological, pharmacological, and genetic approaches, we provide evidence that both Plasmodium and Toxoplasma hijack host cell calpain proteases to facilitate parasite egress. Immunodepletion or inhibition of calpain-1 in hypotonically lysed and resealed erythrocytes prevented the escape of P. falciparum parasites, which was restored by adding purified calpain-1. Similarly, efficient egress of T. gondii from mammalian fibroblasts was blocked by either small interfering RNA– mediated suppression or genetic deletion of calpain activity and could be restored by genetic complementation

Screening the medicines for Malaria Venture "Malaria Box" against the Plasmodium falciparum aminopeptidases, M1, M17 and M18

Malaria is a parasitic disease that remains a global health burden. The ability of the parasite to rapidly develop resistance to therapeutics drives an urgent need for the delivery of new drugs. The Medicines for Malaria Venture have compounds known for their antimalarial ac- tivity, but not necessarily the molecular targets. In this study, we assess the ability of the “MMV 400” compounds to inhibit the activity of three metalloaminopeptidases from Plasmo- dium falciparum, PfA-M1, PfA-M17 and PfM18 AAP. We have developed a multiplex assay system to allow rapid primary screening of compounds against all three metalloaminopepti- dases, followed by detailed analysis of promising compounds. Our results show that there were no PfM18AAP inhibitors, whereas two moderate inhibitors of the neutral aminopepti- dases PfA-M1 and PfA-M17 were identified. Further investigation through structure-activity relationship studies and molecular docking suggest that these compounds are competitive inhibitors with novel binding mechanisms, acting through either non-classical zinc coordina- tion or independently of zinc binding altogether. Although it is unlikely that inhibition of PfA- M1 and/or PfA-M17 is the primary mechanism responsible for the antiplasmodial activity re- ported for these compounds, their detailed characterization, as presented in this work, pave the way for their further optimization as a novel class of dual PfA-M1/PfA-M17 inhibitors uti- lising non-classical zinc binding groups

Root exudate analogues accelerate CO 2 and CH 4 production in tropical peat

Root exudates represent a large and labile carbon input in tropical peatlands, but their contribution to carbon dioxide (CO2) and methane (CH4) production remains poorly understood. Changes in species composition and productivity of peatland plant communities in response to global change could alter both inputs of exudates and associated greenhouse gas emissions. We used manipulative laboratory incubations to assess the extent to which root exudate quantity and chemical composition drives greenhouse gas emissions from tropical peatlands. Peat was sampled from beneath canopy palms (Raphia taedigera) and broadleaved evergreen trees (Campnosperma panamensis) in an ombrotrophic wetland in Panama. Root exudate analogues comprising a mixture of sugars and organic acids were added in solution to peats derived from both species, with CO2 and CH4 measured over time. CO2 and CH4 production increased under most treatments, but the magnitude and duration of the response depended on the composition of the added labile carbon mixture rather than the quantity of carbon added or the botanical origin of the peat. Treatments containing organic acids increased soil pH and altered other soil properties including redox potential but did not affect the activities of extracellular hydrolytic enzymes. CO2 but not CH4 production was found to be linearly related to microbial activity and redox potential. Our findings demonstrate the importance of root exudate composition in regulating greenhouse gas fluxes and propose that in situ plant species changes, particularly those associated with land use change, may account for small scale spatial variation in CO2 and CH4 fluxes due to species specific root exudate compositions

Potent dual inhibitors of Plasmodium falciparum M1 and M17 aminopeptidases through optimization of S1 pocket interactions

Malaria remains a global health problem, and though international efforts for treatment and eradication have made some headway, the emergence of drug-resistant parasites threatens this progress. Antimalarial therapeutics acting via novel mechanisms are urgently required. P. falciparum M1 and M17 are neutral aminopeptidases which are essential for parasite growth and development. Previous work in our group has identified inhibitors capable of dual inhibition of PfA-M1 and PfA-M17, and revealed further regions within the protease S1 pockets that could be exploited in the development of ligands with improved inhibitory activity. Herein, we report the structure-based design and synthesis of novel hydroxamic acid analogues that are capable of potent inhibition of both PfA-M1 and PfA-M17. Furthermore, the developed compounds potently inhibit Pf growth in culture, including the multi-drug resistant strain Dd2. The ongoing development of dual PfA-M1/PfA-M17 inhibitors continues to be an attractive strategy for the design of novel antimalarial therapeutics

Evaluation of anti-biofilm activity of acidic amino acids and synergy with ciprofloxacin on Staphylococcus aureus biofilms

Acidic amino acids, aspartic acid (Asp) and glutamic acid (Glu) can enhance the solubility of many poorly soluble drugs including ciprofloxacin (Cip). One of the mechanisms of resistance within a biofilm is retardation of drug diffusion due to poor penetration across the matrix. To overcome this challenge, this work set to investigate novel counter ion approach with acidic amino acids, which we hypothesised will disrupt the biofilm matrix as well as simultaneously improve drug effectiveness. The anti-biofilm activity of D-Asp and D-Glu was studied on Staphylococcus aureus biofilms. Synergistic effect of combining D-amino acids with Cip was also investigated as a strategy to overcome anti-microbial resistance in these biofilms. Interestingly at equimolar combinations, D-Asp and D-Glu were able to significantly disperse (at 20 mM and 40 mM) established biofilms and inhibit (at 10 mM, 20 mM and 40 mM) new biofilm formation in the absence of an antibiotic. Moreover, our study confirmed L-amino acids also exhibit anti-biofilm activity. The synergistic effect of acidic amino acids with Cip was observed at lower concentration ranges (<40 mM amino acids and <90.54 µM, respectively), which resulted in 96.89% (inhibition) and 97.60% (dispersal) reduction in CFU with exposure to 40 mM amino acids. Confocal imaging indicated that the amino acids disrupt the honeycomb-like extracellular DNA (eDNA) meshwork whilst also preventing its formation

Fingerprinting the Substrate Specificity of M1 and M17 Aminopeptidases of Human Malaria, Plasmodium falciparum

Plasmodium falciparum, the causative agent of human malaria, expresses two aminopeptidases, PfM1AAP and PfM17LAP, critical to generating a free amino acid pool used by the intraerythrocytic stage of the parasite for proteins synthesis, growth and development. These exopeptidases are potential targets for the development of a new class of anti-malaria drugs.To define the substrate specificity of recombinant forms of these two malaria aminopeptidases we used a new library consisting of 61 fluorogenic substrates derived both from natural and unnatural amino acids. We obtained a detailed substrate fingerprint for recombinant forms of the enzymes revealing that PfM1AAP exhibits a very broad substrate tolerance, capable of efficiently hydrolyzing neutral and basic amino acids, while PfM17LAP has narrower substrate specificity and preferentially cleaves bulky, hydrophobic amino acids. The substrate library was also exploited to profile the activity of the native aminopeptidases in soluble cell lysates of P. falciparum malaria.This data showed that PfM1AAP and PfM17LAP are responsible for majority of the aminopeptidase activity in these extracts. These studies provide specific substrate and mechanistic information important for understanding the function of these aminopeptidases and could be exploited in the design of new inhibitors to specifically target these for anti-malaria treatment

Soils of India: historical perspective, classification and recent advances

Derived from a wide range of rocks and minerals, a large variety of soils occur in the Indian subcontinent. Soil-forming factors like climate, vegetation and topography acting for varying periods on a range of rock formations and parent materials, have given rise to different kinds of soil. The National Bureau of Soil Survey and Land Use Planning, Nagpur has developed a database on soils with field and laboratory studies over the last 30 years. This has generated maps and soil information at different scales, showing area and distribution of various soil groups in different agroecological subregions. The 1 : 250,000 scale map shows a threshold soil variation index of 4–5 and 10–25 soil families per m ha for alluvial plains and black soil regions respectively. Progress in basic and fundamental research in Indian soils has been reviewed in terms of soils, their formation related to climate, relief, organisms, parent materials and time

Two-pronged attack: dual inhibition of Plasmodium falciparum M1 and M17 metalloaminopeptidases by a novel series of hydroxamic acid-based inhibitors

Plasmodium parasites, the causative agents of malaria, have developed resistance to most of our current antimalarial therapies, including artemisinin combination therapies which are widely described as our last line of defense. Antimalarial agents with a novel mode of action are urgently required. Two Plasmodium falciparum aminopeptidases, PfA-M1 and PfA-M17, play crucial roles in the erythrocytic stage of infection and have been validated as potential antimalarial targets. Using compound-bound crystal structures of both enzymes, we have used a structure-guided approach to develop a novel series of inhibitors capable of potent inhibition of both PfA-M1 and PfA-M17 activity and parasite growth in culture. Herein we describe the design, synthesis, and evaluation of a series of hydroxamic acid-based inhibitors and demonstrate the compounds to be exciting new leads for the development of novel antimalarial therapeutics

Soil information system: use and potentials in humid and semi-arid tropics

The articles presented in this special section emanated from the researches of consortium members of the National Agricultural Innovative Project (NAIP, Component 4) of the Indian Council of Agricultural Research (ICAR), New Delhi. These researches have helped develop a soil information system (SIS). In view of the changing scenario all over the world, the need of the hour is to get assistance from a host of researchers specialized in soils, crops, geology, geography and information technology to make proper use of the datasets. Equipped with the essential knowledge of data storage and retrieval for management recommendations, these experts should be able to address the issues of land degradation, biodiversity, food security, climate change and ultimately arrive at an appropriate agricultural land-use planning. Moreover, as the natural resource information is an essential prerequisite for monitoring and predicting global environmental change with special reference to climate and land use options, the SIS needs to be a dynamic exercise to accommodate temporal datasets, so that subsequently it should result in the evolution of the soil information technology. The database developed through this NAIP would serve as an example of the usefulness of the Consortium and the research initiative of ICAR involving experts from different fields to find out the potentials of the soils of humid and semi-arid bioclimatic systems of the country

Georeferenced soil information system: assessment of database

Land-use planning is a decision-making process that facilitates the allocation of land to different uses that provide optimal and sustainable benefit. As land-use is shaped by society–nature interaction, in land-use planning different components/facets play a significant role involving soil, water, climate, animal (ruminant/ non-ruminant) and others, including forestry and the environment needed for survival of mankind. At times these components are moderated by human interference. Thus land-use planning being a dynamic phenomenon is not guided by a single factor, but by a complex system working simultaneously,which largely affects the sustainability. To address such issues a National Agricultural Innovation Project (NAIP) on ‘Georeferenced soil information system for land-use planning and monitoring soil and land quality for agriculture’ was undertaken to develop threshold values of land quality parameters for land-use planning through quantitative land evaluation and crop modelling for dominant cropping systems in major agro-ecological sub-regions (AESRs) representing rice–wheat cropping system in the Indo-Gangetic Plains (IGP) and deep-rooted crops in the black soil regions (BSR). To assess the impact of landuse change, threshold land quality indicator values are used. A modified AESR map for agricultural landuse planning is generated for effective land-use planning