Mutations in Transmembrane Domains 1, 4 and 9 of the Plasmodium falciparum Chloroquine Resistance Transporter Alter Susceptibility to Chloroquine, Quinine and Quinidine

Mutations in the Plasmodium falciparum chloroquine (CQ) resistance transporter (PfCRT) can result in verapamil-reversible CQ resistance and altered susceptibility to other antimalarials. PfCRT contains 10 membrane-spanning domains and is found in the digestive vacuole (DV) membrane of intraerythrocytic parasites. The mechanism by which PfCRT mediates CQ resistance is unclear although it is associated with decreased accumulation of drug within the DV. On the permissive background of the P. falciparum 106/1(K76) parasite line, we used single-step drug selection to generate isogenic clones containing unique pfcrt point mutations that resulted in amino acid changes in PfCRT transmembrane domains 1 (C72R, K76N, K76I and K76T) and 9 (Q352K, Q352R). The resulting changes of charge and hydropathy affected quantitative CQ susceptibility and accumulation as well as the stereospecific responses to quinine and quinidine. These results, together with a previously described S163R mutation in transmembrane domain 4, indicate that transmembrane segments 1, 4 and 9 of PfCRT provide important structural components of a substrate recognition and translocation domain. Charge-affecting mutations within these segments may affect the ability of PfCRT to bind different quinoline drugs and determine their net accumulation in the DV. © 2006 The Authors Journal compilation © 2006 Blackwell Publishing Lt

Possible test for CPT invariance with correlated neutral B decays

We study breakdown of $CPT$ symmetry which can occur in the decay process $B \bar B \to l^\pm X^\mp f$ with $f$ being a CP eigenstate. In this process, the standard model expectations for time ordered semi-leptonic and hadronic events, i.e. which of the two decays takes place first, can be altered in the case that there is a violation of the $CPT$ symmetry. To illustrate this possibility, we identify and study several time integrated observables. We find that an experiment with $10^{9}$ $B\bar B$ pairs, has the capability for improving the bound on $CPT$ violating parameter or perhaps observe $CPT$ violation.Comment: Revised version to be published in PR

Azonexus hydrophilus sp nov., a nifH gene-harbouring bacterium isolated from freshwater

Three Gram-negative, non-pigmented, rod-shaped, facultatively aerobic bacterial strains, designated d8-1(T), d8-2 and IMCC1716, were isolated from a freshwater spring sample and a eutrophic freshwater pond. Based on characterization using a polyphasic approach, the three strains showed highly similar phenotypic, physiological and genetic characteristics. All of the strains harboured the nitrogenase gene nifH, but nitrogen-fixing activities could not be detected in nitrogen-free culture media. The three strains shared 99.6-99.7 % 16S rRNA gene sequence similarity and showed 89-100 % DNA-DNA relatedness, suggesting that they represent a single genomic species. Phylogenetic analysis based on 16S rRNA gene sequences showed that strains d8-1(T), d8-2 and IMCC1716 formed a monophyletic branch in the periphery of the evolutionary radiation occupied by the genus Azonexus. Their closest neighbours; were Azonexus caeni Slu-05(T) (96.7-96.8% similarity) and Azonexus fungiphilus BS5-8(T) (96.3-96.6 %). The DNA-DNA relatedness of the novel strains to these two species of the genus Azonexus was less than 70%. The isolates could also be differentiated from recognized members of the genus Azonexus on the basis of phenotypic and biochemical characteristics. It is evident, therefore, that the three strains represent a novel species of the genus Azonexus, for which the name Azonexus hydrophilus sp. nov. is proposed. The type strain is d8-1(T) (=LMG 24005(T)=BCRC 17657 (T))

Distribution and density of the partition function zeros for the diamond-decorated Ising model

Exact renormalization map of temperature between two successive decorated lattices is given, and the distribution of the partition function zeros in the complex temperature plane is obtained for any decoration-level. The rule governing the variation of the distribution pattern as the decoration-level changes is given. The densities of the zeros for the first two decoration-levels are calculated explicitly, and the qualitative features about the densities of higher decoration-levels are given by conjecture. The Julia set associated with the renormalization map is contained in the distribution of the zeros in the limit of infinite decoration level, and the formation of the Julia set in the course of increasing the decoration-level is given in terms of the variations of the zero density.Comment: 8 pages,8figure

Generating oxidation-resistant variants of Bacillus kaustophilus leucine aminopeptidase by substitution of the critical methionine residues with leucine

Bacillus kaustophilus leucine aminopeptidase (bkLAP) was sensitive to oxidative damage by hydrogen peroxide. To improve its oxidative stability, the oxidation-sensitive methionine residues in the enzyme were replaced with leucine by site-directed mutagenesis. The variants, each with an apparent molecular mass of approximately 54 kDa, were overexpressed in recombinant Escherichia coli M15 cells and purified to homogeneity by nickel-chelate chromatography. The specific activity for M282L, M285L, M289L and M321L decreased by more than 43%, while M400L, M426L, M445L, and M485L showed 191, 79, 313, and 103%, respectively, higher activity than the wild-type enzyme. Although the mutations did not cause significant changes in the K-m value, more than 67.8% increase in the value of k(cat)/K-m was observed in the M400L, M426L, M445L and M485L. In the presence of 50 mM H2O2 most variants were more stable with respect to the wild-type enzyme, indicating that the oxidative stability of the enzyme can be improved by engineering the methionine residues

A thermostable leucine aminopeptidase from Bacillus kaustophilus CCRC 11223

Two degenerate primers established from the consensus sequences of bacterial leucine aminopeptidases (LAP) were used to amplify a 360-bp gene fragment from the chromosomal DNA of thermophilic Bacillus kaustophilus CCRC 11223 and the amplified fragment was successfully used as a probe to clone a leucine aminopeptidase (lap) gene from a genomic library of the strain. The gene consists of an open reading frame (ORF) of 1,494 bp and encodes a protein of 497 amino acid residues with a calculated molecular mass of 53.7 kDa. The complete amino acid sequence of the cloned enzyme showed greater than 30% identity with prokaryotic and eukaryotic LAPs. Phylogenetic analysis showed that B. kaustophilus LAP is closely related to the enzyme from Bacillus subtilis and is grouped with the M17 family. His(6)-tagged LAP was generated in Escherichia coli by cloning the coding region into pQE-30 and the recombinant enzyme was purified by nickel-chelate chromatography. The pH and temperature optima for the purified enzyme were 8 and 65degreesC, respectively, and 50% of its activity remained after incubation at 60degreesC for 32 min. The enzyme preferentially hydrolyzed L-leucine-p-nitroanilide (L-Leu-p-NA) followed by Cys derivative

Cluster Spin Glass Distribution Functions in La2−x_{2-x}Srx_xCuO4_4

Signatures of the cluster spin glass have been found in a variety of experiments, with an effective onset temperature $T_{on}$ that is frequency dependent. We reanalyze the experimental results and find that they are characterized by a distribution of activation energies, with a nonzero glass transition temperature $T_g(x)<T_{on}$. While the distribution of activation energies is the same, the distribution of weights depends on the process. Remarkably, the weights are essentially doping independent.Comment: 5 pages, 5 ps figure

Predicting evolution and visualizing high-dimensional fitness landscapes

The tempo and mode of an adaptive process is strongly determined by the structure of the fitness landscape that underlies it. In order to be able to predict evolutionary outcomes (even on the short term), we must know more about the nature of realistic fitness landscapes than we do today. For example, in order to know whether evolution is predominantly taking paths that move upwards in fitness and along neutral ridges, or else entails a significant number of valley crossings, we need to be able to visualize these landscapes: we must determine whether there are peaks in the landscape, where these peaks are located with respect to one another, and whether evolutionary paths can connect them. This is a difficult task because genetic fitness landscapes (as opposed to those based on traits) are high-dimensional, and tools for visualizing such landscapes are lacking. In this contribution, we focus on the predictability of evolution on rugged genetic fitness landscapes, and determine that peaks in such landscapes are highly clustered: high peaks are predominantly close to other high peaks. As a consequence, the valleys separating such peaks are shallow and narrow, such that evolutionary trajectories towards the highest peak in the landscape can be achieved via a series of valley crossingsComment: 12 pages, 7 figures. To appear in "Recent Advances in the Theory and Application of Fitness Landscapes" (A. Engelbrecht and H. Richter, eds.). Springer Series in Emergence, Complexity, and Computation, 201

Chiral symmetry breaking in hot matter

This series of three lectures covers (a) a basic introduction to symmetry breaking in general and chiral symmetry breaking in QCD, (b) an overview of the present status of lattice data and the knowlegde that we have at finite temperature from chiral perturbation theory. (c) Results obtained from the Nambu--Jona-Lasinio model describing static mesonic properties are discussed as well as the bulk thermodynamic quantities. Divergences that are observed in the elastic quark-antiquark scattering cross-section, reminiscent of the phenomenon of critical opalescence in light scattering, is also discussed. (d) Finally, we deal with the realm of systems out of equilibrium, and examine the effects of a medium dependent condensate in a system of interacting quarks.Comment: 62 LaTex pages, incorporating 23 figures. Lectures given at the eleventh Chris-Engelbrecht Summer School in Theoretical Physics, 4-13 February, 1998, to be published by Springer Verla

Effects of built environment on walking among Hong Kong older adults

&nbsp;Key Messages1. Reliable and valid intervieweradministered&nbsp;questionnaires&nbsp;were developed to investigate&nbsp;associations of perceived&nbsp;neighbourhood attributes of&nbsp;Hong Kong older adults with&nbsp;their walking for transportation and recreation.2. Access to and availability of&nbsp;different types of services&nbsp;and destinations, provision offacilities for resting/sitting in&nbsp;the neighbourhood, and easy&nbsp;access to/from residentialbuildings may help maintain an&nbsp;active lifestyle by facilitating&nbsp;walking for transport in the&nbsp;neighbourhood.3. Access to services, indoor places&nbsp;for walking, environmental&nbsp;aesthetics, low traffic, and&nbsp;absence of physical barriers may&nbsp;promote recreational walking