Spectroscopic Signatures of the Superorbital Period in the Neutron Star Binary LMC X-4

We present the first high-resolution X-ray study of emission line variability with superorbital phase in the neutron star binary LMC X-4. Our analysis provides new evidence from X-ray spectroscopy confirming accretion disk precession as the origin of the superorbital period. The spectra, obtained with the Chandra High-Energy Transmission Grating Spectrometer (HETGS) and the XMM-Newton Reflection Grating Spectrometer (RGS), contain a number of emission features, including lines from hydrogen-like and helium-like species of N, O, Ne, and Fe, a narrow O VII RRC, and fluorescent emission from cold Fe. We use the narrow RRC and the He-alpha triplets to constrain the temperature and density of the (photoionized) gas. By comparing spectra from different superorbital phases, we attempt to isolate the contributions to line emission from the accretion disk and the stellar wind. There is also evidence for highly ionized iron redshifted and blueshifted by ~25,000 km/s. We argue that this emission originates in the inner accretion disk, and show that the emission line properties in LMC X-4 are natural consequences of accretion disk precession.Comment: 12 pages, 8 figures, uses emulateap

Unexpectedly high barriers to M–P rotation in tertiary phobane complexes : PhobPR behavior that is commensurate with tBu2PR

The four isomers of 9-butylphosphabicyclo[3.3.1]nonane, s-PhobPBu, where Bu = n-butyl, sec-butyl, isobutyl, tert-butyl, have been prepared. Seven isomers of 9-butylphosphabicyclo[4.2.1]nonane (a5-PhobPBu, where Bu = n-butyl, sec-butyl, isobutyl, tert-butyl; a7-PhobPBu, where Bu = n-butyl, isobutyl, tert-butyl) have been identified in solution; isomerically pure a5-PhobPBu and a7-PhobPBu, where Bu = n-butyl, isobutyl, have been isolated. The σ-donor properties of the PhobPBu ligands have been compared using the JPSe values for the PhobP(═Se)Bu derivatives. The following complexes have been prepared: trans-[PtCl2(s-PhobPR)2] (R = nBu (1a), iBu (1b), sBu (1c), tBu (1d)); trans-[PtCl2(a5-PhobPR)2] (R = nBu (2a), iBu (2b)); trans-[PtCl2(a7-PhobPR)2] (R = nBu (3a), iBu (3b)); trans-[PdCl2(s-PhobPR)2] (R = nBu (4a), iBu (4b)); trans-[PdCl2(a5-PhobPR)2] (R = nBu (5a), iBu (5b)); trans-[PdCl2(a7-PhobPR)2] (R = nBu (6a), iBu (6b)). The crystal structures of 1a–4a and 1b–6b have been determined, and of the ten structures, eight show an anti conformation with respect to the position of the ligand R groups and two show a syn conformation. Solution variable-temperature 31P NMR studies reveal that all of the Pt and Pd complexes are fluxional on the NMR time scale. In each case, two species are present (assigned to be the syn and anti conformers) which interconvert with kinetic barriers in the range 9 to >19 kcal mol–1. The observed trend is that, the greater the bulk, the higher the barrier. The magnitudes of the barriers to M–P bond rotation for the PhobPR complexes are of the same order as those previously reported for tBu2PR complexes. Rotational profiles have been calculated for the model anionic complexes [PhobPR-PdCl3]− using DFT, and these faithfully reproduce the trends seen in the NMR studies of trans-[MCl2(PhobPR)2]. Rotational profiles have also been calculated for [tBu2PR-PdCl3]−, and these show that the greater the bulk of the R group, the lower the rotational barrier: i.e., the opposite of the trend for [PhobPR-PdCl3]−. Calculated structures for the species at the maxima and minima in the M–P rotation energy curves indicate the origin of the restricted rotation. In the case of the PhobPR complexes, it is the rigidity of the bicycle that enforces unfavorable H···Cl clashes involving the Pd–Cl groups with H atoms on the α- or β-carbon in the R substituent and H atoms in 1,3-axial sites within the phosphabicycle

Understanding Mechanisms of Sustained Malaria Transmission in Zambia Through Plasmodium falciparum Genetics

Malaria remains an enormous public health burden, particularly in sub-Saharan Africa where it is among the leading causes of childhood mortality. Recently, renewed global commitment to malaria elimination has laid the groundwork for 35 nations to declare elimination targets and implement elimination programs. Zambia, one such country, is located in southern Africa where malaria control efforts have been challenging. Although Zambia has achieved significant progress towards its targeted elimination deadline of 2021, much work remains. Zambia faces a heterogenous transmission landscape, with regions of low prevalence in the south and regions of high transmission in the northwest, along the international border with the Democratic Republic of the Congo (DRC). To eliminate malaria, Zambia must address the obstacles to malaria control in regions where interventions have been ineffective as well as the barriers to elimination in regions where unknown mechanisms continue to sustain transmission. We examine the barriers to malaria control and elimination in two epidemiologically distinct regions in Zambia. We focus on the utility of P. falciparum genetic methods to draw inferences regarding the mechanisms that continue to sustain malaria transmission in these settings. In southern Zambia, we use microsatellite genotyping to demonstrate that local malaria transmission contributes to the burden of malaria in spite of control measures including reactive case detection. We further identify a region in our study site as a local transmission hotspot, and suggest that this may be an area to prioritize for additional vector control measures. Along the border between Nchelenge District, Zambia and Haut-Katanga Province, DRC, we use amplicon deep sequencing at two diverse P. falciparum loci, Pfama1 and Pfcsp, to infer that cross-border malaria transmission may contribute to the high burden of malaria in this region. We characterize the extent to which this malaria-parasite population is genetically similar to the strain included in the vaccine, RTS,S/AS01. Our analysis indicates that only 5.2% of parasites in this region match the vaccine strain in the C-terminal Pfcsp locus, suggesting that the vaccine may have reduced efficacy in this region. This dissertation demonstrates the value of incorporating parasite genetic analyses into malaria control and elimination efforts

Complete Anopheles funestus mitogenomes reveal an ancient history of mitochondrial lineages and their distribution in southern and central Africa

Anopheles funestus s.s. is a primary vector of malaria in sub-Saharan Africa. Despite its important role in human Plasmodium transmission, evolutionary history, genetic diversity, and population structure of An. funestus in southern and central Africa remains understudied. We deep sequenced, assembled, and annotated the complete mitochondrial genome of An. funestus s.s. for the first time, providing a foundation for further genetic research of this important malaria vector species. We further analyzed the complete mitochondrial genomes of 43 An. funestus s.s. from three sites in Zambia, Democratic Republic of the Congo, and Tanzania. From these 43 mitogenomes we identified 41 unique haplotypes that comprised 567 polymorphic sites. Bayesian phylogenetic reconstruction confirmed the co-existence of two highly divergent An. funestus maternal lineages, herein defined as lineages I and II, in Zambia and Tanzania. The estimated coalescence time of these two mitochondrial lineages is ~500,000 years ago (95% HPD 426,000-594,000 years ago) with subsequent independent diversification. Haplotype network and phylogenetic analysis revealed two major clusters within lineage I, and genetic relatedness of samples with deep branching in lineage II. At this time, data suggest that the lineages are partially sympatric. This study illustrates that accurate retrieval of full mitogenomes of Anopheles vectors enables fine-resolution studies of intraspecies genetic relationships, population differentiation, and demographic history. Further investigations on whether An. funestus mitochondrial lineages represent biologically meaningful populations and their potential implications for malaria vector control are warranted

Therapeutic Efficacy of Artemether-Lumefantrine for Uncomplicated Falciparum Malaria in Northern Zambia.

Artemether-lumefantrine (AL) is a first-line agent for uncomplicated malaria caused by Plasmodium falciparum. The WHO recommends periodic therapeutic efficacy studies of antimalarial drugs for the detection of malaria parasite drug resistance and to inform national malaria treatment policies. We conducted a therapeutic efficacy study of AL in a high malaria transmission region of northern Zambia from December 2014 to July 2015. One hundred children of ages 6 to 59 months presenting to a rural health clinic with uncomplicated falciparum malaria were admitted for treatment with AL (standard 6-dose regimen) and followed weekly for 5 weeks. Parasite counts were taken every 6 hours during treatment to assess parasite clearance. Recurrent episodes during follow-up (n = 14) were genotyped to distinguish recrudescence from reinfection and to identify drug resistance single nucleotide polymorphisms (SNPs) and multidrug resistance protein 1 (mdr1) copy number variation. Day 7 lumefantrine concentrations were measured for correspondence with posttreatment reinfection. All children who completed the parasite clearance portion of the study (n = 94) were microscopy-negative by 72 hours. The median parasite elimination half-life was 2.7 hours (interquartile range: 2.1-3.3). Genotype-corrected therapeutic efficacy was 98.8% (95% CI: 97.6-100). Purported artemisinin and lumefantrine drug resistance SNPs in atp6, 3D7_1451200, and mdr1 were detected but did not correlate with parasite recurrence, nor did day 7 lumefantrine concentrations. In summary, AL was highly effective for the treatment of uncomplicated falciparum malaria in northern Zambia during the study period. The high incidence of recurrent parasitemia was consistent with reinfection due to high, perennial malaria transmission

Organelle-specific expression of subunit ND5 of human complex I (NADH dehydrogenase) alters cation homeostasis in Saccharomyces cerevisiae

The ND5 component of the respiratory complex I is a large, hydrophobic subunit encoded by the mitochondrial genome. Its bacterial homologue, the NDH-1 subunit NuoL, acts as a cation transporter in the absence of other NDH-1 subunits. Mutations in human ND5 are frequently observed in neurodegenerative diseases. Wild type and mutant variants of ND5 fused to GFP or a FLAG peptide were targeted to the endoplasmatic reticulum (ER) or the inner mitochondrial membrane of Saccharomyces cerevisiae, which lacks an endogenous complex I. The localization of ND5 fusion proteins was confirmed by microscopic analyses of S. cerevisiae cells, followed by cellular fractionation and immunostaining. The impact of the expression of ND5 fusion proteins on the growth of S. cerevisiae in the presence and absence of added salts was studied. ER-resident ND5 conferred Li(+) sensitivity to S. cerevisiae, which was lost when the E145V variant of ND5 was expressed. All variants of ND5 tested led to increased resistance of S. cerevisiae at high external concentrations of Na(+) or K(+). The data seem to indicate that ND5 influences the salt homeostasis of S. cerevisiae independent of other complex I subunits, and paves the way for functional studies of mutations found in mitochondrially encoded complex I genes