A neglected disease of humans: a new focus of visceral leishmaniasis in Bakool, Somalia.

Visceral leishmaniasis (VL) was observed in children in Bakool region, Somalia, an area where VL has not been reported before. We describe the extent of the problem in this war- and famine-stricken area. A retrospective analysis was done of all cases admitted to a VL treatment centre between July 2000 and August 2001. Patients with longstanding fever, splenomegaly and a positive direct agglutination test (DAT; titre > 1:3200) were treated as suspected VL cases. A rapid epidemiological and entomological assessment was performed in the area. Species identification was attempted from blood samples by polymerase chain reaction-restriction fragment length polymorphism analysis of cysteine proteinase B genes. In 1 year, 230 serologically-positive cases were diagnosed as VL, and response to therapy was good in 91.6% of the 225 treated with sodium stibogluconate. Parasitological confirmation was attempted and obtained in 2 cases. Parasites were found to be most similar to Sudanese and Ethiopian reference strains of the Leishmania donovani complex. In a serological survey of 161 healthy displaced persons, 15% were positive by the leishmanin skin test and 3 (2%) were positive by the DAT. The sandfly captures showed Phlebotomus martini and P. vansomerenae. VL seems to be a longstanding and serious health problem in Bakool region. Food insecurity might have contributed to the emergence and detection of VL in this area

Fluorescent oxide nanoparticles adapted to active tips for near-field optics

We present a new kind of fluorescent oxide nanoparticles with properties well suited to active-tip based near-field optics. These particles with an average diameter in the range 5-10 nm are produced by Low Energy Cluster Beam Deposition (LECBD) from a YAG:Ce3+ target. They are studied by transmission electron microscopy (TEM), X-ray photoelectron spectroscopy (XPS), classical photoluminescence, cathodoluminescence and near-field scanning optical microscopy (NSOM). Particles of extreme photo-stability as small as 10 nm in size are observed. These emitters are validated as building blocks of active NSOM tips by coating a standard optical tip with a 10 nm thick layer of YAG:Ce3+ particles directly in the LECBD reactor and by subsequently performing NSOM imaging of test surfaces.Comment: Changes made following Referee's comments; added references; one added figure. See story on this article at: http://nanotechweb.org/cws/article/tech/3606

Autocorrelation analysis for the unbiased determination of power-law exponents in single-quantum-dot blinking

We present an unbiased and robust analysis method for power-law blinking statistics in the photoluminescence of single nano-emitters, allowing us to extract both the bright- and dark-state power-law exponents from the emitters' intensity autocorrelation functions. As opposed to the widely-used threshold method, our technique therefore does not require discriminating the emission levels of bright and dark states in the experimental intensity timetraces. We rely on the simultaneous recording of 450 emission timetraces of single CdSe/CdS core/shell quantum dots at a frame rate of 250 Hz with single photon sensitivity. Under these conditions, our approach can determine ON and OFF power-law exponents with a precision of 3% from a comparison to numerical simulations, even for shot-noise-dominated emission signals with an average intensity below 1 photon per frame and per quantum dot. These capabilities pave the way for the unbiased, threshold-free determination of blinking power-law exponents at the micro-second timescale

Gene expression profiling of Leishmania (Leishmania) donovani: overcoming technical variation and exploiting biological variation

Gene expression profiling is increasingly used in the field of infectious diseases for characterization of host, pathogen and the nature of their interaction. The purpose of this study was to develop a robust, standardized method for comparative expression profiling and molecular characterization of Leishmania donovani clinical isolates. The limitations and possibilities associated with expression profiling in intracellular amastigotes and promastigotes were assessed through a series of comparative experiments in which technical and biological parameters were scrutinized. On a technical level, our results show that it is essential to use parasite harvesting procedures that involve minimal disturbance of the parasite's environment in order to ‘freeze' gene expression levels instantly; this is particularly a delicate task for intracellular amastigotes and for specific ‘sensory' genes. On the biological level, we demonstrate that gene expression levels fluctuate during in vitro development of both intracellular amastigotes and promastigotes. We chose to use expression-curves rather than single, specific, time-point measurements to capture this biological variation. Intracellular amastigote protocols need further refinement, but we describe a first generation tool for high-throughput comparative molecular characterization of patients' isolates, based on the changing expression profiles of promastigotes during in vitro differentiatio

Excitons and charged excitons in semiconductor quantum wells

A variational calculation of the ground-state energy of neutral excitons and of positively and negatively charged excitons (trions) confined in a single-quantum well is presented. We study the dependence of the correlation energy and of the binding energy on the well width and on the hole mass. The conditional probability distribution for positively and negatively charged excitons is obtained, providing information on the correlation and the charge distribution in the system. A comparison is made with available experimental data on trion binding energies in GaAs-, ZnSe-, and CdTe-based quantum well structures, which indicates that trions become localized with decreasing quantum well width.Comment: 9 pages, 11 figure

Genomic and metabolomic polymorphism among experimentally selected paromomycin-resistant Leishmania donovani strains

Understanding the mechanism(s) underpinning drug resistance could lead to novel treatments to reverse the increased tolerance of a pathogen. In this study paromomycin (PMM) resistance (PMM-R) was induced in three Nepalese clinical strains of L. donovani, with different inherent susceptibility to antimony drugs (Sb), by step-wise exposure of promastigotes to PMM. Exposure to PMM resulted in the production of mixed populations of parasites even though a single cloned population was used at the start of selection. PMM IC50 values for PMM-R parasites varied between 104-481 µM at the promastigotes stage and 32-195 µM at the intracellular amastigotes stage. PMM resistance was associated with increased resistance to nitric oxide at the amastigote but not the promastigote stage (p < 0.05). This effect was most marked in the Sb-R PMM-R clone, where PMM-R resistance was associated with a significant upregulation in glutathione compared to its WT (p < 0.05) although there was no change in trypanothione (detected in its oxidised form). Interestingly, PMM-R strains showed an increase in either the keto acid derivative of isoleucine (Sb-I PMM-R) or the 2-hydroxy acids derived from arginine and tyrosine (Sb-S PMM-R and Sb-R PMM-R). These results are consistent with the recent finding that upregulation of the branch-chain amino acid aminotransferase and the D-lactate dehydrogenase are linked to PMM-R. In addition, we found that PMM-R was associated with a significant increase in aneuploidy during PMM selection in all the strains, which could allow rapid selection of genetic changes that confer a survival advantage

Directed evolution of artificial repeat proteins as habit modifiers for the morphosynthesis of (111)-terminated gold nanocrystals

Natural biocomposites are shaped by proteins that have evolved to interact with inorganic materials. Protein directed evolution methods which mimic Darwinian evolution have proven highly successful to generate improved enzymes or therapeutic antibodies but have rarely been used to evolve protein–material interactions. Indeed, most reported studies have focused on short peptides and a wide range of oligopeptides with chemical binding affinity for inorganic materials have been uncovered by phage display methods. However, their small size and flexible unfolded structure prevent them from dictating the shape and crystallinity of the growing material. In the present work, a specific set of artificial repeat proteins (αRep), which exhibit highly stable 3D folding with a well-defined hypervariable interacting surface, is selected by directed evolution of a very efficient home-built protein library for their high and selective affinity for the Au(111) surface. The proteins are built from the extendable concatenation of self-compatible repeated motifs idealized from natural HEAT proteins. The high-yield synthesis of Au(111)-faceted nanostructures mediated by these αRep proteins demonstrates their chemical affinity and structural selectivity that endow them with high crystal habit modification performances. Importantly, we further exploit the protein shell spontaneously assembled on the nanocrystal facets to drive protein-mediated colloidal self-assembly and on-surface enzymatic catalysis. Our method constitutes a generic tool for producing nanocrystals with determined faceting, superior biocompatibility and versatile bio-functionalization towards plasmon-based devices and (bio)molecular sensors

Comparison of gene expression patterns among Leishmania braziliensis clinical isolates showing a different in vitro susceptibility to pentavalent antimony

Introduction. Evaluation of Leishmania drug susceptibility depends on in vitro SbV susceptibility assays, which are labour-intensive and may give a biased view of the true parasite resistance. Molecular markers are urgently needed to improve and simplify the monitoring of SbV-resistance. We analysed here the gene expression profile of 21 L. braziliensis clinical isolates in vitro defined as SbV-resistant and -sensitive, in order to identify potential resistance markers. Methods. The differential expression of 13 genes involved in SbV metabolism, oxidative stress or housekeeping functions was analysed during in vitro promastigote growth. Results. Expression profiles were up-regulated for 5 genes only, each time affecting a different set of isolates (mosaic picture of gene expression). Two genes, ODC (ornithine decarboxylase) and TRYR (trypanothione reductase), showed a significantly higher expression rate in the group of SbV-resistant compared to the group of SbV-sensitive parasites (P<0·01). However, analysis of individual isolates showed both markers to explain only partially the drug resistance. Discussion. Our results might be explained by (i) the occurrence of a pleiotropic molecular mechanism leading to the in vitro SbV resistance and/or (ii) the existence of different epi-phenotypes not revealed by the in vitro SbV susceptibility assays, but interfering with the gene expression pattern