A Family of Putative Receptor-Adenylate Cyclases from Leishmania donovani

Leishmania parasites are exposed to pronounced changes in their environment during their life cycle as they migrate from the sandfly midgut to the insect proboscis and then into the phagolysosomes of the vertebrate macrophages. The developmental transformations that produce each life cycle stage of the parasite may be signaled in part by binding of environmental ligands to receptors which mediate transduction of extracellular signals. We have identified a family of five clustered genes in Leishmania donovani which may encode signal transduction receptors. The coding regions of two of these genes, designated rac-A and rac-B, have been sequenced and shown to code for proteins with an NH2-terminal hydrophilic domain, an intervening putative transmembrane segment, and a COOH-terminal domain that has high sequence identity to the catalytic domain from adenylate cyclases in other eukaryotes. We have expressed the receptor-adenylate cyclase protein (RAC)-A protein in Xenopus oocytes and demonstrated that it functions as an adenylate cyclase. Although RAC-B exhibits no catalytic activity when expressed in oocytes, co-expression of RAC-A and RAC-B negatively regulates the adenylate cyclase activity of RAC-A, suggesting that these two proteins interact in the membrane. Furthermore, a truncated version of RAC-A functions as a dominant negative mutant that inhibits the catalytic activity of the wild type receptor. The rac-A and rac-B genes encode developmentally regulated mRNAs which are expressed in the insect stage but not in the mammalian host stage of the parasite life cycle

Improved Synthesis for Modular Ascarosides Uncovers Biological Activity

A versatile synthesis of modular ascarosides, a family of signaling molecules from Caenorhabditis elegans and other nematodes, via hydrogenolysis of a cyclic sulfate derived from methyl-α-l-rhamnopyranoside is reported. The route enables selective introduction of different side chains at the 1, 2, and 4 positions of the sugar, as demonstrated for ascarosides from C. elegans and Pristionchus pacificus. Bioassays with synthetic samples of 4′-tigloyl ascaroside mbas#3 revealed its role as an avoidance or dispersal signal

Electromagnetic phenomena in heterogeneous media: Effective properties and local behavior

The purpose of this paper is the use of a mathematical homogenization approach based on the multiple scale expansion theory for modeling the electromagnetic phenomena arising in heterogeneous media under an imposed magnetic flux. The attention is focused on the analysis and discussion of the merits and limits of this theoretical approach in reproducing not only the effective macroscopic properties but also the local behavior, under a wide frequency range and considering different constitutive and geometrical parameters. The results show that the proposed method is able to predict local and integral physical quantities, ranging from a substantially global behavior in the whole media to significantly localized effects determined by the microscopic structure

KHARON Is an essential cytoskeletal protein involved in the trafficking of flagellar membrane proteins and cell division in African trypanosomes

African trypanosomes and related kinetoplastid parasites selectively traffic specific membrane proteins to the flagellar membrane, but the mechanisms for this trafficking are poorly understood. We show here that KHARON, a protein originally identified in Leishmania parasites, interacts with a putative trypanosome calcium channel and is required for its targeting to the flagellar membrane. KHARON is located at the base of the flagellar axoneme, where it likely mediates targeting of flagellar membrane proteins, but is also on the subpellicular microtubules and the mitotic spindle. Hence, KHARON is probably a multifunctional protein that associates with several components of the trypanosome cytoskeleton. RNA interference-mediated knockdown of KHARON mRNA results in failure of the calcium channel to enter the flagellar membrane, detachment of the flagellum from the cell body, and disruption of mitotic spindles. Furthermore, knockdown of KHARON mRNA induces a lethal failure of cytokinesis in both bloodstream (mammalian host) and procyclic (insect vector) life cycle stages, and KHARON is thus critical for parasite viability

Analysis of the reciprocal wear testing of Aluminum AA1050 processed by a novel mechanical nanostructuring technique

This research aims to investigate the impact of a novel technique in mechanical nanostructuring on the wear resistance of materials. This technique with the name of High Pressure Torsion Extrusion (HPTE) can produce bulk nanostructured materials with enhanced mechanical properties. Results of microstructural analysis and microhardness testing showed significant enhancement in materials after HPTE. Microstructural characterization by using Electron Back-Scattered Diffraction (EBSD) method illustrated the presence of Ultra-Fine Grained (UFG) materials in the specimens Analysis of the wear by implementing reciprocal wear testing revealed that the amount of displaced volume markedly decreased after processing. This change in the wear behavior can be explained by referring to the hardness increase and the reduction of plasticity in materials which confined the plastic shearing and diminished the built-up edge around the wear track

Differences at brain SPECT between depressed females with and without adult ADHD and healthy controls: etiological considerations

<p>Abstract</p> <p>Background</p> <p>Comorbidity between Attention Deficit Hyperactivity Disorder (ADHD) and mood disorders is common. Alterations of the cerebellum and frontal regions have been reported in neuro-imaging studies of ADHD and major depression.</p> <p>Methods</p> <p>Thirty chronically depressed adult females of whom 16 had scores below, and 14 scores above, cut-offs on the 25-items Wender Utah Retrospective Scale (WURS-25) and the Wender-Reimherr Adult Attention Deficit Disorder Scale (WRAADDS) were divided into subgroups designated "Depression" and "Depression + ADHD", respectively. Twenty-one of the patients had some audiological symptom, tinnitus and/or hearing impairment. The patients were investigated with other rating scales and ^99mTc-HMPAO SPECT. Controls for ^99mTc-HMPAO SPECT were 16 healthy females. SPECT was analyzed by both statistical parametric mapping (SPM2) and the computerized brain atlas (CBA). Discriminant analysis was performed on the volumes of interest generated by the CBA, and on the scores from rating scales with the highest group differences.</p> <p>Results</p> <p>The mean score of a depression rating scale (MADRS-S) was significantly lower in the "Depression" subgroup compared to in the "Depression + ADHD" subgroup. There was significantly decreased tracer uptake within the bilateral cerebellum at both SPM and CBA in the "Depression + ADHD" subgroup compared to in the controls. No decrease of cerebellar tracer uptake was observed in "Depression". Significantly increased tracer uptake was found at SPM within some bilateral frontal regions (Brodmann areas 8, 9, 10, 32) in the "Depression + ADHD" subgroup compared to in "Depression". An accuracy of 100% was obtained for the discrimination between the patient groups when thalamic uptake was used in the analysis along with scores from Socialization and Impulsivity scales.</p> <p>Conclusion</p> <p>The findings confirm the previous observation of a cerebellar involvement in ADHD. Higher bilateral frontal ^99mTc-HMPAO uptake in "Depression + ADHD" compared to in "Depression" indicate a difference between these subgroups. ^99mTc-HMPAO uptake mechanisms are discussed.</p

Arsenic transport by zebrafish aquaglyceroporins

<p>Abstract</p> <p>Background</p> <p>Arsenic is one of the most ubiquitous toxins and endangers the health of tens of millions of humans worldwide. It is a mainly a water-borne contaminant. Inorganic trivalent arsenic (As^III) is one of the major species that exists environmentally. The transport of As^IIIhas been studied in microbes, plants and mammals. Members of the aquaglyceroporin family have been shown to actively conduct As^IIIand its organic metabolite, monomethylarsenite (MAs^III). However, the transport of As^IIIand MAs^IIIin in any fish species has not been characterized.</p> <p>Results</p> <p>In this study, five members of the aquaglyceroporin family from zebrafish (<it>Danio rerio</it>) were cloned, and their ability to transport water, glycerol, and trivalent arsenicals (As^IIIand MAs^III) and antimonite (Sb^III) was investigated. Genes for at least seven aquaglyceroporins have been annotated in the zebrafish genome project. Here, five genes which are close homologues to human AQP3, AQP9 and AQP10 were cloned from a zebrafish cDNA preparation. These genes were named <it>aqp3, aqp3l, aqp9a, aqp9b </it>and <it>aqp10 </it>according to their similarities to the corresponding human AQPs. Expression of <it>aqp9a, aqp9b</it>, <it>aqp3, aqp3l </it>and <it>aqp10 </it>in multiple zebrafish organs were examined by RT-PCR. Our results demonstrated that these aquaglyceroporins exhibited different tissue expression. They are all detected in more than one tissue. The ability of these five aquaglyceroporins to transport water, glycerol and the metalloids arsenic and antimony was examined following expression in oocytes from <it>Xenopus leavis</it>. Each of these channels showed substantial glycerol transport at equivalent rates. These aquaglyceroporins also facilitate uptake of inorganic As^III, MAs^IIIand Sb^III. Arsenic accumulation in fish larvae and in different tissues from adult zebrafish was studied following short-term arsenic exposure. The results showed that liver is the major organ of arsenic accumulation; other tissues such as gill, eye, heart, intestine muscle and skin also exhibited significant ability to accumulate arsenic. The zebrafish larvae also accumulate considerable amounts of arsenic.</p> <p>Conclusion</p> <p>This is the first molecular identification of fish arsenite transport systems and we propose that the extensive expression of the fish aquaglyceroporins and their ability to transport metalloids suggests that aquaglyceroporins are the major pathways for arsenic accumulation in a variety of zebrafish tissues. Uptake is one important step of arsenic metabolism. Our results will contribute to a new understanding of aquatic arsenic metabolism and will support the use of zebrafish as a new model system to study arsenic associated human diseases.</p

Increasing pattern recognition accuracy for chemical sensing by evolutionary based drift compensation

Artificial olfaction systems, which mimic human olfaction by using arrays of gas chemical sensors combined with pattern recognition methods, represent a potentially low-cost tool in many areas of industry such as perfumery, food and drink production, clinical diagnosis, health and safety, environmental monitoring and process control. However, successful applications of these systems are still largely limited to specialized laboratories. Sensor drift, i.e., the lack of a sensor's stability over time, still limits real in dustrial setups. This paper presents and discusses an evolutionary based adaptive drift-correction method designed to work with state-of-the-art classification systems. The proposed approach exploits a cutting-edge evolutionary strategy to iteratively tweak the coefficients of a linear transformation which can transparently correct raw sensors' measures thus mitigating the negative effects of the drift. The method learns the optimal correction strategy without the use of models or other hypotheses on the behavior of the physical chemical sensors