Changes in Purkinje cell firing and gene expression precede behavioral pathology in a mouse model of SCA2.

Spinocerebellar ataxia type 2 (SCA2) is an autosomal dominantly inherited disorder, which is caused by a pathological expansion of a polyglutamine (polyQ) tract in the coding region of the ATXN2 gene. Like other ataxias, SCA2 most overtly affects Purkinje cells (PCs) in the cerebellum. Using a transgenic mouse model expressing a full-length ATXN2(Q127)-complementary DNA under control of the Pcp2 promoter (a PC-specific promoter), we examined the time course of behavioral, morphologic, biochemical and physiological changes with particular attention to PC firing in the cerebellar slice. Although motor performance began to deteriorate at 8 weeks of age, reductions in PC number were not seen until after 12 weeks. Decreases in the PC firing frequency first showed at 6 weeks and paralleled deterioration of motor performance with progression of disease. Transcription changes in several PC-specific genes such as Calb1 and Pcp2 mirrored the time course of changes in PC physiology with calbindin-28 K changes showing the first small, but significant decreases at 4 weeks. These results emphasize that in this model of SCA2, physiological and behavioral phenotypes precede morphological changes by several weeks and provide a rationale for future studies examining the effects of restoration of firing frequency on motor function and prevention of future loss of PCs

Finite-temperature behavior of the Bose polaron

We consider a mobile impurity immersed in a Bose gas at finite temperature. Using perturbation theory valid for weak coupling between the impurity and the bosons, we derive analytical results for the energy and damping of the impurity for low and high temperatures, as well as for temperatures close to the critical temperature $T_c$ for Bose-Einstein condensation. These results show that the properties of the impurity vary strongly with temperature. In particular, the energy exhibits a non-monotonic behavior close to $T_c$, and the damping rises sharply close to $T_c$. We argue that this behaviour is generic for impurities immersed in an environment undergoing a phase transition that breaks a continuous symmetry. Finally, we discuss how these effects can be detected experimentally.Comment: 10 pages and 6 figure

A positive feedback loop linking enhanced mGluR function and basal calcium in spinocerebellar ataxia type 2

Metabotropic glutamate receptor 1 (mGluR1) function in Purkinje neurons (PNs) is essential for cerebellar development and for motor learning and altered mGluR1 signaling causes ataxia. Downstream of mGluR1, dysregulation of calcium homeostasis has been hypothesized as a key pathological event in genetic forms of ataxia but the underlying mechanisms remain unclear. We find in a spinocerebellar ataxia type 2 (SCA2) mouse model that calcium homeostasis in PNs is disturbed across a broad range of physiological conditions. At parallel fiber synapses, mGluR1-mediated excitatory postsynaptic currents (EPSCs) and associated calcium transients are increased and prolonged in SCA2 PNs. In SCA2 PNs, enhanced mGluR1 function is prevented by buffering [Ca 2+ ] at normal resting levels while in wildtype PNs mGluR1 EPSCs are enhanced by elevated [Ca 2+ ]. These findings demonstrate a deleterious positive feedback loop involving elevated intracellular calcium and enhanced mGluR1 function, a mechanism likely to contribute to PN dysfunction and loss in SCA2

A reinforcing circuit action of extrasynaptic GABAA receptor modulators on cerebellar granule cell inhibition.

GABAA receptors (GABARs) are the targets of a wide variety of modulatory drugs which enhance chloride flux through GABAR ion channels. Certain GABAR modulators appear to acutely enhance the function of δ subunit-containing GABAR subtypes responsible for tonic forms of inhibition. Here we identify a reinforcing circuit mechanism by which these drugs, in addition to directly enhancing GABAR function, also increase GABA release. Electrophysiological recordings in cerebellar slices from rats homozygous for the ethanol-hypersensitive (α6100Q) allele show that modulators and agonists selective for δ-containing GABARs such as THDOC, ethanol and THIP (gaboxadol) increased the frequency of spontaneous inhibitory postsynaptic currents (sIPSCs) in granule cells. Ethanol fails to augment granule cell sIPSC frequency in the presence of glutamate receptor antagonists, indicating that circuit mechanisms involving granule cell output contribute to ethanol-enhancement of synaptic inhibition. Additionally, GABAR antagonists decrease ethanol-induced enhancement of Golgi cell firing. Consistent with a role for glutamatergic inputs, THIP-induced increases in Golgi cell firing are abolished by glutamate receptor antagonists. Moreover, THIP enhances the frequency of spontaneous excitatory postsynaptic currents in Golgi cells. Analyses of knockout mice indicate that δ subunit-containing GABARs are required for enhancing GABA release in the presence of ethanol and THIP. The limited expression of the GABAR δ subunit protein within the cerebellar cortex suggests that an indirect, circuit mechanism is responsible for stimulating Golgi cell GABA release by drugs selective for extrasynaptic isoforms of GABARs. Such circuit effects reinforce direct actions of these positive modulators on tonic GABAergic inhibition and are likely to contribute to the potent effect of these compounds as nervous system depressants

Mutagenesis of Trichoderma reesei endoglucanase I: impact of expression host on activity and stability at elevated temperatures.

BackgroundTrichoderma reesei is a key cellulase source for economically saccharifying cellulosic biomass for the production of biofuels. Lignocellulose hydrolysis at temperatures above the optimum temperature of T. reesei cellulases (~50°C) could provide many significant advantages, including reduced viscosity at high-solids loadings, lower risk of microbial contamination during saccharification, greater compatibility with high-temperature biomass pretreatment, and faster rates of hydrolysis. These potential advantages motivate efforts to engineer T. reesei cellulases that can hydrolyze lignocellulose at temperatures ranging from 60-70°C.ResultsA B-factor guided approach for improving thermostability was used to engineer variants of endoglucanase I (Cel7B) from T. reesei (TrEGI) that are able to hydrolyze cellulosic substrates more rapidly than the recombinant wild-type TrEGI at temperatures ranging from 50-70°C. When expressed in T. reesei, TrEGI variant G230A/D113S/D115T (G230A/D113S/D115T Tr_TrEGI) had a higher apparent melting temperature (3°C increase in Tm) and improved half-life at 60°C (t1/2 = 161 hr) than the recombinant (T. reesei host) wild-type TrEGI (t1/2 = 74 hr at 60°C, Tr_TrEGI). Furthermore, G230A/D113S/D115T Tr_TrEGI showed 2-fold improved activity compared to Tr_TrEGI at 65°C on solid cellulosic substrates, and was as efficient in hydrolyzing cellulose at 60°C as Tr_TrEGI was at 50°C. The activities and stabilities of the recombinant TrEGI enzymes followed similar trends but differed significantly in magnitude depending on the expression host (Escherichia coli cell-free, Saccharomyces cerevisiae, Neurospora crassa, or T. reesei). Compared to N.crassa-expressed TrEGI, S. cerevisiae-expressed TrEGI showed inferior activity and stability, which was attributed to the lack of cyclization of the N-terminal glutamine in Sc_TrEGI and not to differences in glycosylation. N-terminal pyroglutamate formation in TrEGI expressed in S. cerevisiae was found to be essential in elevating its activity and stability to levels similar to the T. reesei or N. crassa-expressed enzyme, highlighting the importance of this ubiquitous modification in GH7 enzymes.ConclusionStructure-guided evolution of T. reesei EGI was used to engineer enzymes with increased thermal stability and activity on solid cellulosic substrates. Production of TrEGI enzymes in four hosts highlighted the impact of the expression host and the role of N-terminal pyroglutamate formation on the activity and stability of TrEGI enzymes

Pressure Raman effects and internal stress in network glasses

Raman scattering from binary GexSe1-x glasses under hydrostatic pressure shows onset of a steady increase in the frequency of modes of corner-sharing GeSe4 tetrahedral units when the external pressure P exceeds a threshold value Pc. The threshold pressure Pc(x) decreases with x in the 0.15 < x < 0.20 range, nearly vanishes in the 0.20 < x < 0.25 range, and then increases in the 0.25 < x < 1/3 range. These Pc(x) trends closely track those in the non-reversing enthalpy, DHnr(x), near glass transitions (Tgs), and in particular, both DHnr(x) and Pc(x) vanish in the reversibility window (0.20 < x < 0.25). It is suggested that Pc provides a measure of stress at the Raman active units; and its vanishing in the reversibility window suggests that these units are part of an isostatically rigid backbone. Isostaticity also accounts for the non-aging behavior of glasses observed in the reversibility window

Gene co-expression network analysis for identifying modules and functionally enriched pathways in SCA2

Spinocerebellar ataxia type 2 (SCA2) is an autosomal dominant neurodegenerative disease caused by CAG repeat expansion in the ATXN2 gene. The repeat resides in an encoded region of the gene resulting in polyglutamine (polyQ) expansion which has been assumed to result in gain of function, predominantly, for the ATXN2 protein. We evaluated temporal cerebellar expression profiles by RNA sequencing of ATXN2Q127 mice versus wild-type (WT) littermates. ATXN2Q127 mice are characterized by a progressive motor phenotype onset, and have progressive cerebellar molecular and neurophysiological (Purkinje cell firing frequency) phenotypes. Our analysis revealed previously uncharacterized early and progressive abnormal patterning of cerebellar gene expression. Weighted Gene Coexpression Network Analysis revealed four gene modules that were significantly correlated with disease status, composed primarily of genes associated with GTPase signaling, calcium signaling and cell death. Of these genes, few overlapped with differentially expressed cerebellar genes that we identified in Atxn2−/− knockout mice versus WT littermates, suggesting that loss-of-function is not a significant component of disease pathology. We conclude that SCA2 is a disease characterized by gain of function for ATXN2

Green Manufacturing - An overview

Data on energy consumption, global warming, and carbon di-oxide levels in the environment, industrial pollution, and population is growing extensively which means there are more and more challenges with less sustainability, which leads to think about Green Manufacturing. Consumerism driven consumption in developed countries and population in developing countries is leading to tremendous demand for goods and services. Fulfilling such ever increasing global demands is putting up the pressures on limited resources viz materials and energy. Unfortunately rate of depletion of material and energy has outgrown the rate at which nature recovers stroke restores them back. With this ever widening gap the day is not far for our future generations to strive for basic needs. To overcome this challenge society must embrace sustainable methodologies and practices. Being the source of maximum industrial pollution manufacturing industries must lead the way toward Green Manufacturing. The paper gives an overview of Green Manufacturing, drivers of Green manufacturing, and case studies of Green manufacturing

Functional Imaging Using InGaAs/GaAs Photorefractive Multiple Quantum Wells

doi:10.1063/1.1994722 http://link.aip.org/link/?APCPCS/772/1581/1We propose the use of an InGaAs/GaAs photorefractive quantum well (PRQW) as an adaptive beam combiner for holographic optical coherence imaging applications. Holograms have been observed by using a diode laser and an interferometer. A weaker quantum confined exciton leads to the saturation of electroabsorption and hence diffraction, under a high external electric field, in the InGaAs PRQW. A careful choice of external electric field modulation seems to reduce this effect. We examine several characteristics that govern the use of an InGaAs PRQW in a functional imaging system