High-Content Chemical and RNAi Screens for Suppressors of Neurotoxicity in a Huntington's Disease Model

To identify Huntington's Disease therapeutics, we conducted high-content small molecule and RNAi suppressor screens using a Drosophila primary neural culture Huntingtin model. Drosophila primary neurons offer a sensitive readout for neurotoxicty, as their neurites develop dysmorphic features in the presence of mutant polyglutamine-expanded Huntingtin compared to nonpathogenic Huntingtin. By tracking the subcellular distribution of mRFP-tagged pathogenic Huntingtin and assaying neurite branch morphology via live-imaging, we identified suppressors that could reduce Huntingtin aggregation and/or prevent the formation of dystrophic neurites. The custom algorithms we used to quantify neurite morphologies in complex cultures provide a useful tool for future high-content screening approaches focused on neurodegenerative disease models. Compounds previously found to be effective aggregation inhibitors in mammalian systems were also effective in Drosophila primary cultures, suggesting translational capacity between these models. However, we did not observe a direct correlation between the ability of a compound or gene knockdown to suppress aggregate formation and its ability to rescue dysmorphic neurites. Only a subset of aggregation inhibitors could revert dysmorphic cellular profiles. We identified lkb1, an upstream kinase in the mTOR/Insulin pathway, and four novel drugs, Camptothecin, OH-Camptothecin, 18β-Glycyrrhetinic acid, and Carbenoxolone, that were strong suppressors of mutant Huntingtin-induced neurotoxicity. Huntingtin neurotoxicity suppressors identified through our screen also restored viability in an in vivo Drosophila Huntington's Disease model, making them attractive candidates for further therapeutic evaluation.National Institutes of Health (U.S.) (grant R01 EB007042)National Institutes of Health (U.S.

Neurexins and Neuroligins: Recent Insights from Invertebrates

During brain development, each neuron must find and synapse with the correct pre- and postsynaptic partners. The complexity of these connections and the relatively large distances some neurons must send their axons to find the correct partners makes studying brain development one of the most challenging, and yet fascinating disciplines in biology. Furthermore, once the initial connections have been made, the neurons constantly remodel their dendritic and axonal arbours in response to changing demands. Neurexin and neuroligin are two cell adhesion molecules identified as important regulators of this process. The importance of these genes in the development and modulation of synaptic connectivity is emphasised by the observation that mutations in these genes in humans have been associated with cognitive disorders such as Autism spectrum disorders, Tourette syndrome and Schizophrenia. The present review will discuss recent advances in our understanding of the role of these genes in synaptic development and modulation, and in particular, we will focus on recent work in invertebrate models, and how these results relate to studies in mammals

Percepção dos consumidores relativamente à carne de bovino: critérios de escolha a partir de grupos de discussão

Nas economias desenvolvidas, a segurança dos alimentos, o bem‑estar animal e o ambiente têm vindo a assumir relevância nas preocupações dos consumidores. Procurámos saber, através da metodologia dos grupos de discussão (focus groups) realizados em duas cidades de Portugal, Lisboa e Porto, as preocupações e perceções dos consumidores relativamente a estes atributos, os quais são essencialmente acreditados. Este conhecimento é fundamental para a aplicação posterior de métodos de preferências declaradas. Foi ainda possível obter, com base em exercícios de seleção entre carnes diferenciadas, e através de um modelo logit, intervalos de preços passíveis de serem utilizados posteriormente na definição de cenários em experiências de escolha (método de preferências declaradas) -----ABSTRACT-----In developed economies, food safety, animal welfare and the environment have become relevant consumers’ concerns. We conducted several focus groups in two Portuguese cities, Lisboa and Porto, in order to get participants perceptions and concerns for these attributes, which are essentially credence attributes. This knowledge is critical for the subsequent application of stated preference methods. It was also possible to estimate, based on exercises of selection among different meats, and through a logit model, price ranges that could be used in the definition of scenarios for choice experiments (stated preference method)info:eu-repo/semantics/publishedVersio

Evolution of microstructure and crystallographic texture during dissimilar friction stir welding of duplex stainless steel to low carbon-manganese structural steel

Electron backscattered diffraction (EBSD) was used to analyze the evolution of microstructure and crystallographic texture during friction stir welding of dissimilar type 2205 duplex stainless steel (DSS) to type S275 low carbon-manganese structural steel. The results of microstructural analyses show that the temperature in the center of stirred zone reached temperatures between Ac 1 and Ac 3 during welding, resulting in a minor ferrite-to-austenite phase transformation in the S275 steel, and no changes in the fractions of ferrite and austenite in the DSS. Temperatures in the thermomechanically affected and shoulder-affected zones of both materials, in particular toward the root of the weld, did not exceed the Ac 1 of S275 steel. The shear generated by the friction between the material and the rotating probe occurred in austenitic/ferritic phase field of the S275 and DSS. In the former, the transformed austenite regions of the microstructure were transformed to acicular ferrite, on cooling, while the dual-phase austenitic/ferritic structure of the latter was retained. Studying the development of crystallographic textures with regard to shear flow lines generated by the probe tool showed the dominance of simple shear components across the whole weld in both materials. The ferrite texture in S275 steel was dominated by D 1, D 2, E, E¯ , and F, where the fraction of acicular ferrite formed on cooling showed a negligible deviation from the texture for the ideal shear texture components of bcc metals. The ferrite texture in DSS was dominated by D 1, D 2, I, I¯ , and F, and that of austenite was dominated by the A, A¯ , B, and B¯ of the ideal shear texture components for bcc and fcc metals, respectively. While D 1, D 2, and F components of the ideal shear texture are common between the ferrite in S275 steel and that of dual-phase DSS, the preferential partitioning of strain into the ferrite phase of DSS led to the development of I and I¯ components in DSS, as opposed to E and E¯ in the S275 steel. The formations of fine and ultrafine equiaxed grains were observed in different regions of both materials that are believed to be due to strain-induced continuous dynamic recrystallization (CDRX) in ferrite of both DSS and S275 steel, and discontinuous dynamic recrystallization (DDRX) in austenite phase of DSS

Cardiopoietic cell therapy for advanced ischemic heart failure: results at 39 weeks of the prospective, randomized, double blind, sham-controlled CHART-1 clinical trial

Cardiopoietic cells, produced through cardiogenic conditioning of patients' mesenchymal stem cells, have shown preliminary efficacy. The Congestive Heart Failure Cardiopoietic Regenerative Therapy (CHART-1) trial aimed to validate cardiopoiesis-based biotherapy in a larger heart failure cohort

Processing and structure of in situ Fe-Al alloys produced by gas tungsten arc welding

Iron aluminide weld overlays are being investigated for corrosion and erosion protection of boiler tubes in low NOx burners. The primary objective of the research is to identify overlay compositions which can be deposited in a crack-free condition and provide corrosion protection in moderately reducing environments. In the current phase of work, Fe-Al alloy weld overlays were produced by depositing commercially pure aluminum wire on to low carbon steel substrates using Gas Tungsten Arc Welding. A systematic variation of the wire feed speed and current, two major factors affecting dilution, resulted in a variation in aluminum contents of the welds ranging from 3--42 wt% aluminum. The aluminum content was observed to increase with wire feed speed and a decrease in the current. The aluminum content was also found to affect the cracking susceptibility of the overlays. At 10wt% aluminum, few to no cracks were observed in the deposits. Above this value, cracking was prevalent throughout the weld. In addition, two types of microstructures were found correlating to different concentrations of aluminum. A homogeneous matrix with second phase particles consisting of coarse columnar grains was found for low aluminum concentrations. With higher aluminum contents, a two-phase constituent was observed to surround primary dendrites growing from the substrate. The transition of the microstructures occurred between 24 and 32 wt% Al

Cermet composite thermal spray coatings for erosion and corrosion protection in combustion environments of advanced coal-fired boilers. Semi-annual technical progress report, February 1996--July 1996

Present coal-fired boiler environments remain hostile to the materials of choice since corrosion and erosion can be a serious problem in certain regions of the boiler. Recently, the Clean Air Act Amendment is requiring electric power plants to reduce NO{sub x}, emissions to the environment. To reduce NO{sub x}, emissions, new low NO{sub x}, combustors are utilized which burn fuel with a substoichiometric amount of oxygen (i.e., low oxygen partial pressure). In these low NO{sub x} environments, H{sub 2}S gas is a major source of sulfur. Due to the sulfidation process, corrosion rates in reducing parts of boilers have increased significantly and existing boiler tube materials do not always provide adequate corrosion resistance. Combined attack due to corrosion and erosion is a concern because of the significantly increased operating costs which result in material failures. One method to combat corrosion and erosion in coal-fired boilers is to apply coatings to the components subjected to aggressive environments. Thermal spray coatings, a cermet composite comprised of hard ceramic phases of oxide and/or carbide in a metal binder, have been used with some success as a solution to the corrosion and erosion problems in boilers. However, little is known on the effect of the volume fraction, size, and shape of the hard ceramic phase on the erosion and corrosion resistance of the thermally sprayed coatings. It is the objective of this research to investigate metal matrix composite (cermet) coatings in order to determine the optimum ceramic/metal combination that will give the best erosion and corrosion resistance in new advanced coal-fired boilers