Microtubules as antiparasitic drug targets

Background: Parasitic diseases including malaria, leishmaniasis and schistosomiasis take a terrible toll of human life, health and productivity, especially in tropical and subtropical regions, and are also highly significant in animal health worldwide. Antiparasitic drugs are the mainstays of control of most of these diseases, but in many cases current therapies are inadequate and in some the situation is deteriorating because of drug resistance. Microtubules, as essential components of almost all eukaryotic cells, are proven drug targets in many helminth diseases and show promise as targets for the development of new antiprotozoal drugs. Objective: This article reviews the chemistry of the microtubule inhibitors in current use and under investigation as antiparasitic agents, their activities against the major parasites and their mechanisms of action. New directions in both inhibitor chemistry and biological evaluation are discussed. Conclusions: The most promising immediate avenues for discovery and design appear to lie in development of novel benzimidazoles for helminth parasites and compounds based on antimitotic herbicides for protozoal parasites. New understanding from functional genomics, structural biology and microtubular imaging will help accelerate the development of completely novel antiparasitic drugs targeting microtubules.

Missing the targets and being misunderstood: Child phonology and interactive problems

Effects of vowel variation on interaction are considered, with particular relevance to their role in conversational breakdown. The effect of speaker knowledge and experience is noted as a variable in developmental progress which must inform profiling decisions, and the need for appropriate taxonomies of speech varieties is emphasized as a precursor to clinical and educational assessments. It is noted, too, that a shared sociolinguistic background between speaker and listener does not always resolve difficulties arising from non-target realizations, casting some doubt on ideas that assessors always possess a guaranteed sense of phonological variability and its effects. Hence, an informed understanding of phonological variation, rather than merely awareness that such variation exists, is advocated.

Interactions between the budding yeast IQGAP homologue Iqg1p and its targets revealed by a split-EGFP bimolecular fluorescence complementation assay.

A split-EGFP based bimolecular fluorescence complementation (BiFC) assay has been used to detect interactions between the Saccharomyces cerevisiae cytoskeletal scaffolding protein Iqg1p and three targets: myosin essential light chain (Mlc1p), calmodulin (Cmd1p) and the small GTPase Cdc42p. The format of the BiFC assay used ensures that the proteins are expressed at wild type levels thereby avoiding artefacts due to overexpression. This is the first direct in vivo detection of these interactions; in each case, the complex is localised to discrete regions of the yeast cytoplasm. The labelling with EGFP fragments results in changes in growth kinetics, cell size and budding frequency. This is partly due to the reassembled EGFP locking the complexes into essentially permanent interactions. The consequences of this for Iqg1p interactions and BiFC assays in general are discussed. (c) 2008 International Federation for Cell Biology. Published by Elsevier Ltd. All rights reserved.

New molecular targets in radiotherapy: DNA damage signalling and repair in targeted and non-targeted cells

Ionising radiation plays a key role in therapy due to its ability to directly induce DNA damage, in particular DNA double-strand breaks leading to cell death. Cells have multiple repair pathways which attempt to maintain genomic stability. DNA repair proteins have become key targets for therapy, using small molecule inhibitors, in combination with radiation and or chemotherapeutic agents as a means of enhancing cell killing. Significant advances in our understanding of the response of cells to radiation exposures has come from the observation of non-targeted effects where cells respond via mechanisms other than those which are a direct consequence of energy-dependent DNA damage. Typical of these is bystander signalling where cells respond to the fact that their neighbours have been irradiated. Bystander cells show a DNA damage response which is distinct from directly irradiated cells. In bystander cells, ATM- and Rad3-related (ATR) protein kinase-dependent signalling in response to stalled replication forks is an early event in the DNA damage response. The ATM protein kinase is activated downstream of ATR in bystander cells. This offers the potential for differential approaches for the modulation of bystander and direct effects with repair inhibitors which may impact on the response of tumours and on the protection of normal tissues during radiotherapy. (C) 2009 Elsevier B.V. All rights reserved.

Radiation-Pressure Acceleration of Ion Beams from Nanofoil Targets: The Leaky Light-Sail Regime

A new ion radiation-pressure acceleration regime, the "leaky light sail," is proposed which uses sub-skin-depth nanometer foils irradiated by circularly polarized laser pulses. In the regime, the foil is partially transparent, continuously leaking electrons out along with the transmitted laser field. This feature can be exploited by a multispecies nanofoil configuration to stabilize the acceleration of the light ion component, supplementing the latter with an excess of electrons leaked from those associated with the heavy ions to avoid Coulomb explosion. It is shown by 2D particle-in-cell simulations that a monoenergetic proton beam with energy 18 MeV is produced by circularly polarized lasers at intensities of just 10(19) W/cm(2). 100 MeV proton beams are obtained by increasing the intensities to 2 x 10(20) W/cm(2).

Effect of self-generated magnetic fields on fast-electron beam divergence in solid targets

The collimating effect of self-generated magnetic fields on fast-electron transport in solid aluminium targets irradiated by ultra-intense, picosecond laser pulses is investigated in this study. As the target thickness is varied in the range of 25 mu m to 1.4 mm, the maximum energies of protons accelerated from the rear surface are measured to infer changes in the fast-electron density and therefore the divergence of the fast-electron beam transported through the target. Purely ballistic spreading of the fast-electrons would result in a much faster decrease in the maximum proton energy with increasing target thickness than that measured. This implies that some degree of 'global' magnetic pinching of the fast-electrons occurs, particularly for thick (>400 mu m) targets. Numerical simulations of electron transport are in good agreement with the experimental data and show that the pinching effect of the magnetic field in thin targets is significantly reduced due to disruption of the field growth by refluxing fast-electrons.

Carbon ion acceleration from thin foil targets irradiated by ultrahigh-contrast, ultraintense laser pulses

In this study, ion acceleration from thin planar target foils irradiated by ultrahigh-contrast (10(10)), ultrashort (50 fs) laser pulses focused to intensities of 7 x 10(20) W cm(-2) is investigated experimentally. Target normal sheath acceleration (TNSA) is found to be the dominant ion acceleration mechanism when the target thickness is >= 50 nm and laser pulses are linearly polarized. Under these conditions, irradiation at normal incidence is found to produce higher energy ions than oblique incidence at 35 degrees with respect to the target normal. Simulations using one-dimensional (1D) boosted and 2D particle-in-cell codes support the result, showing increased energy coupling efficiency to fast electrons for normal incidence. The effects of target composition and thickness on the acceleration of carbon ions are reported and compared to calculations using analytical models of ion acceleration.

Guiding of Relativistic Electron Beams in Solid Targets by Resistively Controlled Magnetic Fields

Guided transport of a relativistic electron beam in solid is achieved experimentally by exploiting the strong magnetic fields created at the interface of two metals of different electrical resistivities. This is of substantial relevance to the Fast Ignitor approach to fusion energy production [M. Tabak et al., Phys. Plasmas 12, 057305 (2005)], since it allows the electron deposition to be spatially tailored-thus adding substantial design flexibility and preventing inefficiencies due to electron beam spreading. In the experiment, optical transition radiation and thermal emission from the target rear surface provide a clear signature of the electron confinement within a high resistivity tin layer sandwiched transversely between two low resistivity aluminum slabs. The experimental data are found to agree well with numerical simulations.

Activated oncogenic pathways and therapeutic targets in extranodal nasal-type NK/T cell lymphoma revealed by gene expression profiling

We performed comprehensive genome-wide gene expression profiling (GEP) of extranodal nasal-type natural killer/T-cell lymphoma (NKTL) using formalin-fixed, paraffin-embedded tissue (n = 9) and NK cell lines (n = 5) in comparison with normal NK cells, with the objective of understanding the oncogenic pathways involved in the pathogenesis of NKTL and to identify potential therapeutic targets. Pathway and network analysis of genes differentially expressed between NKTL and normal NK cells revealed significant enrichment for cell cycle-related genes and pathways, such as PLK1, CDK1, and Aurora-A. Furthermore, our results demonstrated a pro-proliferative and anti-apoptotic phenotype in NKTL characterized by activation of Myc and nuclear factor kappa B (NF-kappa B), and deregulation of p53. In corroboration with GEP findings, a significant percentage of NKTLs (n = 33) overexpressed c-Myc (45.4%), p53 (87.9%), and NF-kappa B p50 (67.7%) on immunohistochemistry using a tissue microarray containing 33 NKTL samples. Notably, overexpression of survivin was observed in 97% of cases. Based on our findings, we propose a model of NKTL pathogenesis where deregulation of p53 together with activation of Myc and NF-kappa B, possibly driven by EBV LMP-1, results in the cumulative up-regulation of survivin. Down-regulation of survivin with Terameprocol (EM-1421, a survivin inhibitor) results in reduced cell viability and increased apoptosis in tumour cells, suggesting that targeting survivin may be a potential novel therapeutic strategy in NKTL. Copyright (C) 2011 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.

Targets of genome copy number reduction in primary breast cancers identified by integrative genomics

The identification of specific oncogenes and tumor suppressor genes in regions of recurrent aneuploidy is a major challenge of molecular cancer research. Using both oligonucleotide single-nucleotide polymorphism and mRNA expression arrays, we integrated genomic and transcriptional information to identify and prioritize candidate cancer genes in regions of increased and decreased chromosomal copy number in a cohort of primary breast cancers. Confirming the validity of this approach, several regions of previously-known copy number (CN) alterations in breast cancer could be successfully reidentified. Focusing on regions of decreased CN, we defined a prioritized list of eighteen candidate genes, which included ARPIN, FBNI, and LZTSI, previously shown to be associated with cancers in breast or other tissue types, and novel genes such as P29, MORF4LI, and TBCID5. One such gene, the RUNX3 transcription factor, was selected for further study. We show that RUNX3 is present at reduced CNs in proportion to the rest of the tumor genome and that RUNX3 CN reductions can also be observed in a breast cancer series from a different center. Using tissue microarrays, we demonstrate in an independent cohort of over 120 breast tissues that RUNX3 protein is expressed in normal breast epithelium but not fat and stromal tissue, and widely down-regulated in the majority of breast cancers (> 85%). In vitro, RUNX3 overexpression suppressed the invasive potential of MDA-MB-231 breast cancer cells in a matrigel assay. Our results demonstrate the utility of integrative genomic approaches to identify novel potential cancer-related genes in primary tumors. This article contains Supplementary Material available at http:// www.interscience.wiley.com/jpages/1045-2257/suppmat. (c) 2006 Wiley-Liss, Inc.

Breit interaction in dielectronic recombination of hydrogenlike uranium

Absolute rate coefficients for dielectronic recombination (DR) of H-like U91+ ions have been measured. The electron-ion merged-beam technique at a heavy-ion storage ring was employed using a stochastically cooled ion beam. Thereby, the previously accessible electron-ion collision energies could be greatly extended to the range 63-90 keV. High-resolution DR spectra were measured covering all KLL and KLM resonances. For the resonance strengths, excellent agreement between relativistic theory and experiment is found only if the Breit contribution to the electron-electron interaction is included in the calculations. For the KL1/2L1/2 and KL1/2M1/2 groups the Breit contribution amounts to 44% of their total resonance strengths.