The plasmodium lactate/H+ transporter PfFNT is essential and druggable in vivo

Malaria parasites in the blood stage express a single transmembrane transport protein for the release of the glycolytic end product l-lactate/H(+) from the cell. This transporter is a member of the strictly microbial formate-nitrite transporter (FNT) family and a novel putative drug target. Small, drug-like FNT inhibitors potently block lactate transport and kill Plasmodium falciparum parasites in culture. The protein structure of Plasmodium falciparum FNT (PfFNT) in complex with the inhibitor has been resolved and confirms its previously predicted binding site and its mode of action as a substrate analog. Here, we investigated the mutational plasticity and essentiality of the PfFNT target on a genetic level, and established its in vivo druggability using mouse malaria models. We found that, besides a previously identified PfFNT G107S resistance mutation, selection of parasites at 3 x IC(50) (50% inhibitory concentration) gave rise to two new point mutations affecting inhibitor binding: G21E and V196L. Conditional knockout and mutation of the PfFNT gene showed essentiality in the blood stage, whereas no phenotypic defects in sexual development were observed. PfFNT inhibitors mainly targeted the trophozoite stage and exhibited high potency in P. berghei- and P. falciparum-infected mice. Their in vivo activity profiles were comparable to that of artesunate, demonstrating strong potential for the further development of PfFNT inhibitors as novel antimalarials

Complex microwave conductivity of Na-DNA powders

We report the complex microwave conductivity, $\sigma=\sigma_1-i\sigma_2$, of Na-DNA powders, which was measured from 80 K to 300 K by using a microwave cavity perturbation technique. We found that the magnitude of $\sigma_1$ near room temperature was much larger than the contribution of the surrounding water molecules, and that the decrease of $\sigma_1$ with decreasing temperature was sufficiently stronger than that of the conduction of counterions. These results clearly suggest that the electrical conduction of Na-DNA is intrinsically semiconductive.Comment: 16 pages, 7 figure

The anomaly of the oxygen bond-bending mode at 320 cm−1^{-1} and the additional absorption peak in the c-axis infrared conductivity of underdoped YBa2_{2}Cu3_{3}O7−δ_{7-\delta} single crystals revisited by ellipsometricmeasurements

We have performed ellipsometric measurements of the far-infrared c-axis dielectric response of underdoped YBa$_{2}$Cu$_{3}$O$_{7-\delta}$ single crystals. Here we report a detailed analysis of the temperature-dependent renormalization of the oxygen bending phonon mode at 320 cm$^{-1}$ and the formation of the additional absorption peak around 400-500 cm$^{-1}$. For a strongly underdoped YBa$_{2}$Cu$_{3}$O$_{6.5}$ crystal with T$_{c}$=52 K we find that, in agreement with previous reports based on conventional reflection measurements, the gradual onset of both features occurs well above T$_{c}$ at T*$\sim$150 K. Contrary to some of these reports, however, our data establish that the phonon anomaly and the formation of the additional peak exhibit very pronounced and steep changes right at T$_{c}$. For a less underdoped YBa$_{2}$Cu$_{3}$O$_{6.75}$ crystal with T$_{c}$=80 K, the onset temperature of the phonon anomaly almost coincides with T$_{c}$. Also in contrast to some previous reports, we find for both crystals that a sizeable fraction of the spectral weight of the additional absorption peak cannot be accounted for by the spectral-weight loss of the phonon modes but instead arises from a redistribution of the electronic continuum. Our ellipsometric data are consistent with a model where the bilayer cuprate compounds are treated as a superlattice of intra- and inter-bilayer Josephson-junctions

The catalytic subunit of Plasmodium falciparum casein kinase 2 is essential for gametocytogenesis

Casein kinase 2 (CK2) is a pleiotropic kinase phosphorylating substrates in different cellular compartments in eukaryotes. In the malaria parasite Plasmodium falciparum, PfCK2 is vital for asexual proliferation of blood-stage parasites. Here, we applied CRISPR/Cas9-based gene editing to investigate the function of the PfCK2alpha catalytic subunit in gametocytes, the sexual forms of the parasite that are essential for malaria transmission. We show that PfCK2alpha localizes to the nucleus and cytoplasm in asexual and sexual parasites alike. Conditional knockdown of PfCK2alpha expression prevented the transition of stage IV into transmission-competent stage V gametocytes, whereas the conditional knockout of pfck2a completely blocked gametocyte maturation already at an earlier stage of sexual differentiation. In summary, our results demonstrate that PfCK2alpha is not only essential for asexual but also sexual development of P. falciparum blood-stage parasites and encourage studies exploring PfCK2alpha as a potential target for dual-active antimalarial drugs

The 3-phosphoinositide-dependent protein kinase 1 is an essential upstream activator of protein kinase A in malaria parasites

Cyclic adenosine monophosphate (cAMP)-dependent protein kinase A (PKA) signalling is essential for the proliferation of Plasmodium falciparum malaria blood stage parasites. The mechanisms regulating the activity of the catalytic subunit PfPKAc, however, are only partially understood, and PfPKAc function has not been investigated in gametocytes, the sexual blood stage forms that are essential for malaria transmission. By studying a conditional PfPKAc knockdown (cKD) mutant, we confirm the essential role for PfPKAc in erythrocyte invasion by merozoites and show that PfPKAc is involved in regulating gametocyte deformability. We furthermore demonstrate that overexpression of PfPKAc is lethal and kills parasites at the early phase of schizogony. Strikingly, whole genome sequencing (WGS) of parasite mutants selected to tolerate increased PfPKAc expression levels identified missense mutations exclusively in the gene encoding the parasite orthologue of 3-phosphoinositide-dependent protein kinase-1 (PfPDK1). Using targeted mutagenesis, we demonstrate that PfPDK1 is required to activate PfPKAc and that T189 in the PfPKAc activation loop is the crucial target residue in this process. In summary, our results corroborate the importance of tight regulation of PfPKA signalling for parasite survival and imply that PfPDK1 acts as a crucial upstream regulator in this pathway and potential new drug target

Isothermal Microcalorimetry, a New Tool to Monitor Drug Action against Trypanosoma brucei and Plasmodium falciparum

Isothermal microcalorimetry is an established tool to measure heat flow of physical, chemical or biological processes. The metabolism of viable cells produces heat, and if sufficient cells are present, their heat production can be assessed by this method. In this study, we investigated the heat flow of two medically important protozoans, Trypanosoma brucei rhodesiense and Plasmodium falciparum. Heat flow signals obtained for these pathogens allowed us to monitor parasite growth on a real-time basis as the signals correlated with the number of viable cells. To showcase the potential of microcalorimetry for measuring drug action on pathogenic organisms, we tested the method with three antitrypanosomal drugs, melarsoprol, suramin and pentamidine and three antiplasmodial drugs, chloroquine, artemether and dihydroartemisinin, each at two concentrations on the respective parasite. With the real time measurement, inhibition was observed immediately by a reduced heat flow compared to that in untreated control samples. The onset of drug action, the degree of inhibition and the time to death of the parasite culture could conveniently be monitored over several days. Microcalorimetry is a valuable element to be added to the toolbox for drug discovery for protozoal diseases such as human African trypanosomiasis and malaria. The method could probably be adapted to other protozoan parasites, especially those growing extracellularly

Correlation between the Josephson coupling energy and the condensation energy in bilayer cuprate superconductors

We review some previous studies concerning the intra-bilayer Josephson plasmons and present new ellipsometric data of the c-axis infrared response of almost optimally doped Bi_{2}Sr_{2}CaCu_{2}O_{8}. The c-axis conductivity of this compound exhibits the same kind of anomalies as that of underdoped YBa_{2}Cu_{3}O_{7-delta}. We analyze these anomalies in detail and show that they can be explained within a model involving the intra-bilayer Josephson effect and variations of the electric field inside the unit cell. The Josephson coupling energies of different bilayer compounds obtained from the optical data are compared with the condensation energies and it is shown that there is a reasonable agreement between the values of the two quantities. We argue that the Josephson coupling energy, as determined by the frequency of the intra-bilayer Josephson plasmon, represents a reasonable estimate of the change of the effective c-axis kinetic energy upon entering the superconducting state. It is further explained that this is not the case for the estimate based on the use of the simplest ``tight-binding'' sum rule. We discuss possible interpretations of the remarkable agreement between the Josephson coupling energies and the condensation energies. The most plausible interpretation is that the interlayer tunneling of the Cooper pairs provides the dominant contribution to the condensation energy of the bilayer compounds; in other words that the condensation energy of these compounds can be accounted for by the interlayer tunneling theory. We suggest an extension of this theory, which may also explain the high values of T_{c} in the single layer compounds Tl_{2}Ba_{2}CuO_{6} and HgBa_{2}CuO_{4}, and we make several experimentally verifiable predictions.Comment: 16 pages (including Tables) and 7 figures; accepted for publication in Physical Review

Measurement of the branching ratios of the Z0 into heavy quarks

We measure the hadronic branching ratios of the Z0 boson into heavy quarks: Rb=Gamma(Z0->bb)/Gamma(Z0->hadrons) and Rc=Gamma(Z0->cc/Gamma(Z0->hadrons) using a multi-tag technique. The measurement was performed using about 400,000 hadronic Z0 events recorded in the SLD experiment at SLAC between 1996 and 1998. The small and stable SLC beam spot and the CCD-based vertex detector were used to reconstruct bottom and charm hadron decay vertices with high efficiency and purity, which enables us to measure most efficiencies from data. We obtain, Rb=0.21604 +- 0.00098(stat.) +- 0.00073(syst.) -+ 0.00012(Rc) and, Rc= 0.1744 +- 0.0031(stat.) +- 0.0020(syst.) -+ 0.0006(Rb)Comment: 37 pages, 8 figures, to be submitted to Phys. Rev. D version 2: changed title to ratios, used common D production fractions for Rb and Rc and corrected Zgamma interference. Identical to PRD submissio

Direct Measurements of A_b and A_c using Vertex/Kaon Charge Tags at SLD

Exploiting the manipulation of the SLC electron-beam polarization, we present precise direct measurements of the parity violation parameters A_c and A_b in the Z boson - c quark and Z boson - b quark coupling. Quark/antiquark discrimination is accomplished via a unique algorithm that takes advantage of the precise SLD CCD vertex detector, employing the net charge of displaced vertices as well as the charge of kaons that emanate from those vertices. From the 1996-98 sample of 400,000 Z decays, produced with an average beam polarization of 73.4%, we find A_c = 0.673 +/- 0.029 (stat.) +/- 0.023 (syst.) and A_b = 0.919 +/- 0.018 (stat.) +/- 0.017 (syst.).Comment: 11 pages, 2 figures, 2 tables, to be submitted to Physical Review Letters; version 2 reflects changes suggested by the refere

Search for time-dependent B0s - B0s-bar oscillations using a vertex charge dipole technique

We report a search for B0s - B0s-bar oscillations using a sample of 400,000 hadronic Z0 decays collected by the SLD experiment. The analysis takes advantage of the electron beam polarization as well as information from the hemisphere opposite that of the reconstructed B decay to tag the B production flavor. The excellent resolution provided by the pixel CCD vertex detector is exploited to cleanly reconstruct both B and cascade D decay vertices, and tag the B decay flavor from the charge difference between them. We exclude the following values of the B0s - B0s-bar oscillation frequency: Delta m_s < 4.9 ps-1 and 7.9 < Delta m_s < 10.3 ps-1 at the 95% confidence level.Comment: 18 pages, 3 figures, replaced by version accepted for publication in Phys.Rev.D; results differ slightly from first versio