Biochemical characterization and chemical inhibition o PfATP4-associated Na+-ATPase activity in Plasmodium falciparum membranes

The antimalarial activity of chemically diverse compounds, including the clinical candidate cipargamin, has been linked to the ATPase PfATP4 in the malaria-causing parasite Plasmodium falciparum. The characterization of PfATP4 has been hampered by the inability thus far to achieve its functional expression in a heterologous system. Here, we optimized a membrane ATPase assay to probe the function of PfATP4 and its chemical sensitivity. We found that cipargamin inhibited the Na+-dependent ATPase activity present in P. falciparum membranes from WT parasites and that its potency was reduced in cipargamin-resistant PfATP4-mutant parasites. The cipargamin-sensitive fraction of membrane ATPase activity was inhibited by all 28 of the compounds in the "Malaria Box" shown previously to disrupt ion regulation in P. falciparum in a cipargamin-like manner. This is consistent with PfATP4 being the direct target of these compounds. Characterization of the cipargamin-sensitive ATPase activity yielded data consistent with PfATP4 being a Na+ transporter that is sensitive to physiologically relevant perturbations of pH, but not of [K+] or [Ce2+]. With an apparent K-m for ATP of 0.2 mm and an apparent K-m for Na+ of 16 -17 mm, the protein is predicted to operate at below its half-maximal rate under normal physiological conditions, allowing the rate of Na+ efflux to increase in response to an increase in cytosolic [Na+]. In membranes from a cipargamin-resistant PfATP4-mutant line, the apparent K-m for Na+ is slightly elevated. Our study provides new insights into the biochemical properties and chemical sensitivity of an important new antimalarial drug target.This work was supported by an Australian Research Council Discovery Early Career Researcher Award (DE160101035 to A. M. L.), an Australian Research Council Linkage Project Grant (LP150101226 to K. K.), and a National Health and Medical Research Council Project Grant (1042272 to K. K.). The authors declare that they have no conflicts of interest with the contents of this article

A Complete Scheme of Ionization Cooling for a Muon Collider

A complete scheme for production and cooling a muon beam for three specified muon colliders is presented. Parameters for these muon colliders are given. The scheme starts with the front end of a proposed neutrino factory that yields bunch trains of both muon signs. Emittance exchange cooling in slow helical lattices reduces the longitudinal emittance until it becomes possible to merge the trains into single bunches, one of each sign. Further cooling in all dimensions is applied to the single bunches in further slow helical lattices. Final transverse cooling to the required parameters is achieved in 50 T solenoids using high Tc superconductor at 4 K. Preliminary simulations of each element are presented.Comment: 3 pages, 6 figure

1H-NMR metabolite profiles of different strains of Plasmodium falciparum

Although efforts to understand the basis for inter-strain phenotypic variation in the most virulent malaria species, Plasmodium falciparum, have benefited from advances in genomic technologies, there have to date been few metabolomic studies of this parasite. Using 1H-NMR spectroscopy, we have compared the metabolite profiles of red blood cells infected with different P. falciparum strains. These included both chloroquine-sensitive and chloroquine-resistant strains, as well as transfectant lines engineered to express different isoforms of the chloroquine-resistance-conferring pfcrt (P. falciparum chloroquine resistance transporter). Our analyses revealed strain-specific differences in a range of metabolites. There was marked variation in the levels of the membrane precursors choline and phosphocholine, with some strains having >30-fold higher choline levels and >5-fold higher phosphocholine levels than others. Chloroquineresistant strains showed elevated levels of a number of amino acids relative to chloroquine-sensitive strains, including an approximately 2-fold increase in aspartate levels. The elevation in amino acid levels was attributable to mutations in pfcrt. Pfcrt-linked differences in amino acid abundance were confirmed using alternate extraction and detection (HPLC) methods. Mutations acquired to withstand chloroquine exposure therefore give rise to significant biochemical alterations in the parasite

The RFOFO Ionization Cooling Ring for Muons

Practical ionization cooling rings could lead to lower cost or improved performance in neutrino factory or muon collider designs. The ring modeled here uses realistic three-dimensional fields. The performance of the ring compares favorably with the linear cooling channel used in the second US Neutrino Factory Study. The normalized 6D emittance of an ideal ring is decreased by a factor of approximately 240, compared with a factor of only 15 for the linear channel. We also examine such \textit{real-world} effects as windows on the absorbers and rf cavities and leaving empty lattice cells for injection and extraction. For realistic conditions the ring decreases the normalized 6D emittance by a factor of 49.Comment: 27 pages, 18 figures and 5 tables. Submitted to Phys. Rev. ST-A

Risk Stratification in Women Enrolled in the Acute Decompensated Heart Failure National Registry Emergency Module (ADHERE-EM)

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/75587/1/j.1553-2712.2008.00030.x.pd

Diverse mutational pathways converge on saturable chloroquine transport via the malaria parasite's chloroquine resistance transporter

Mutations in the chloroquine resistance transporter (PfCRT) are the primary determinant of chloroquine (CQ) resistance in the malaria parasite Plasmodium falciparum. A number of distinct PfCRT haplotypes, containing between 4 and 10 mutations, have give

Muon Colliders

Muon Colliders have unique technical and physics advantages and disadvantages when compared with both hadron and electron machines. They should thus be regarded as complementary. Parameters are given of 4 TeV and 0.5 TeV high luminosity \mumu colliders, and of a 0.5 TeV lower luminosity demonstration machine. We discuss the various systems in such muon colliders, starting from the proton accelerator needed to generate the muons and proceeding through muon cooling, acceleration and storage in a collider ring. Problems of detector background are also discussed.Comment: 28 pages, with 12 postscript figures. To be published Proceedings of the 9th Advanced ICFA Beam Dynamics Workshop, AIP Pres