EAE TCR motifs and antigen recognition in myelin basic protein-induced anterior uveitis in Lewis rats

T cells infiltrating the iris/ciliary body of Lewis rats with anterior uveitis (AU) that had been induced by myelin basic protein (MBP) immunization were previously found to share surface markers common to the T cells that cause experimental autoimmune encephalomyelitis (EAE). To determine whether these AU-associated T cells are in fact the same as those that infiltrate the central nervous system to cause EAE, we examined TCR V gene expression in T cells infiltrating the anterior chamber in rats with AU. As with EAE, we found a biased expression of Vbeta8.2 and Valpha2 in the iris/ciliary body and, although one would expect an influx of nonspecific inflammatory T cells, these biases were still evident at the peak of AU. An analysis of the TCR Vbeta8.2 and Valpha2 sequences derived from the iris/ciliary body demonstrated the presence of the same complementarity determining region 3 motifs found in MBP-specific T cells that are pathogenic for EAE and found in T cells derived from the central nervous system of rats with EAE. Finally, T cells isolated from the iris/ciliary body of rats with AU were found to proliferate in a specific fashion to MBP Ags. Thus, it appears that MBP-specific T cells are pathogenic for AU as well as EAE in the Lewis rat. In addition, the long-term presence of this highly restricted MBP response in the iris/ciliary body indicates that distinct immunoregulatory mechanisms exist in the environment of the eye. This provides an interesting model with which to address questions pertaining to the nature of T cells infiltrating the eye and their regulation during EAE and other systemic diseases.     

PRMT 3, a type I protein arginine N-methyltransferase that differs from PRMT1 in its oligomerization, subcellular localization, substrate specificity, and regulation

Methylation is one of the many post-translational modifications that modulate protein function. Although asymmetric NG,NG-dimethylation of arginine residues in glycine-arginine-rich domains of eucaryotic proteins, catalyzed by type I protein arginine N-methyltransferases (PRMT), has been known for some time, members of this enzyme class have only recently been cloned. The first example of this type of enzyme, designated PRMT1, cloned because of its ability to interact with the mammalian TIS21 immediate-early protein, was then shown to have protein arginine methyltransferase activity. We have now isolated rat and human cDNA orthologues that encode proteins with substantial sequence similarity to PRMT1. A recombinant glutathione S-transferase (GST) fusion product of this new rat protein, named PRMT3, asymmetrically dimethylates arginine residues present both in the designed substrate GST-GAR and in substrate proteins present in hypomethylated extracts of a yeast rmt1 mutant that lacks type I arginine methyltransferase activity; PRMT3 is thus a functional type I protein arginine N-methyltransferase. However, rat PRMT1 and PRMT3 glutathione S-transferase fusion proteins have distinct enzyme specificities for substrates present in both hypomethylated rmt1 yeast extract and hypomethylated RAT1 embryo cell extract. TIS21 protein modulates the enzymatic activity of recombinant GST-PRMT1 fusion protein but not the activity of GST-PRMT3. Western blot analysis of gel filtration fractions suggests that PRMT3 is present as a monomer in RAT1 cell extracts. In contrast, PRMT1 is present in an oligomeric complex. Immunofluorescence analysis localized PRMT1 predominantly to the nucleus of RAT1 cells. In contrast, PRMT3 is predominantly cytoplasmic.     

Fast full‐color reflective display via photoluminescent enhancement

Reflective displays are advantageous in applications requiring low power or daylight readability. However, there are no low‐cost reflective technologies capable of displaying bright colors. By employing photoluminescence to more efficiently use ambient light, we created a prototype display that provides bright, full color in a simple, low‐cost architecture. This prototype includes a novel electrokinetic shutter, a layer that incorporates patterned luminescent red, green, and blue sub‐pixel elements, and a novel optical out‐coupling scheme. The luminescent elements convert otherwise‐wasted portions of the incident spectrum to light in the desired color band, resulting in improved color saturation and lightness. This prototype provides a color gamut that is superior to competing reflective display technologies that utilize color filters in single‐layer side‐by‐side sub‐pixel architectures. The current prototype is capable of switching in <0.5 s; future displays based on an alternative electro‐optic shutter technology should achieve video rate operation. A transflective version of this technology has also been prototyped. The transflective version utilizes its backlight with a power efficiency that is at least three times that of a conventional liquid crystal display. These photoluminescence‐based technologies enable a host of applications ranging from low‐power mobile products and retail pricing signage to daylight readable signage for outdoor advertising segments.     

Robust pose invariant face recognition using coupled latent space discriminant analysis

We propose a novel pose-invariant face recognition approach which we call Discriminant Multiple Coupled Latent Subspace framework. It finds the sets of projection directions for different poses such that the projected images of the same subject in different poses are maximally correlated in the latent space. Discriminant analysis with artificially simulated pose errors in the latent space makes it robust to small pose errors caused due to a subject’s incorrect pose estimation. We do a comparative analysis of three popular latent space learning approaches: Partial Least Squares (PLSs), Bilinear Model (BLM) and Canonical Correlational Analysis (CCA) in the proposed coupled latent subspace framework. We experimentally demonstrate that using more than two poses simultaneously with CCA results in better performance. We report state-of-the-art results for pose-invariant face recognition on CMU PIE and FERET and comparable results on MultiPIE when using only four fiducial points for alignment and intensity features.     

Computer simulations of time-dependent suppression of EOF by polymer coatings

We use molecular dynamics simulations in order to investigate the time evolution of the effect of adsorbed polymer coatings on the electro-osmotic flow (EOF) in a capillary. Weakly adsorbed coatings show no time-dependent performance, but they do not strongly reduce the EOF. On the other hand, strongly adsorbed coatings made of longer polymer chains are often quenched in non-equilibrium conformations that can strongly reduce the EOF over extremely long periods of time. For intermediate adsorption strengths, we observe that the EOF increases as a function of time due to the relaxation of the coating layer. The concentration of polymers in solution and the length of the polymer chains also affect the time-dependence of the EOF. These results show that the quality of electrophoretic separations can depend on the waiting time between the formation of the coating and the beginning of the separation. We conclude by suggesting experimental tests of our predictions.     

Vortex rings and the superfluid λ-transition

A vortex-ring formulation of the₄He -transition provides a clear physical picture of the superfluid transition. The thermally excited rings are dipoles which orient in an applied flow field, and their net backflow cancels part of the applied flow, reducing the superfluid density. At the -point rings of infinite size drive the superfluid density to zero as a power law of the reduced temperature, with an exponent of 0.672. By fitting to the experimental superfluid amplitude the core energy of the smallest rings is found to be 6.1K at T and their diameter is 2.3 Å. It is proposed that these can be identified as the roton excitations of the Landau model. The vortex theory also yields new insights into topics such as boundary effects at a wall, finite-size effects, and the dynamics of the transition.     

Whose responsibility is IT (information technology) management?

Line managers are increasingly assuming responsibility for planning, building, and running information systems that affect their operations. This is forcing organizations to evaluate how they allocate IT decision-making responsibilities. This paper describes a conceptual framework and an intervention process that can help firms devise and implement an effective IT management architecture. The authors illustrate their methods with real world examples.     

Steric stabilization of Stöber silica dispersions using organosilanes

Organosilanes of the general formula R _x Si(OR)_4–x (where R is an alkyl group and R = CH₃ or CH₂CH₃) were used to sterically stabilize hexane dispersions of submicrometre silica spheres. The dispersions were characterized according to sediment volume results. For 0.5m silica particles, the sediment density increased by more than a factor of three up to 50 to 55% of theoretical in the presence of organosilanes with 12 or more carbons in the R group. Solid-state¹³C nuclear magnetic resonance was used to characterize the powder-dispersant interaction; this technique can distinguish between carbons in the R group of the organosilane and residual organic groups in the silica. Scanning electron micrographs of filter compacts were used to further characterize the dispersions and indicated the presence of primary particles as well as small agglomerates.     

Validation of a Gas Chromatography-Mass Spectrometry Method for the Measurement of the Redox State Metabolic Ratios Lactate/Pyruvate and β-Hydroxybutyrate/Acetoacetate in Biological Samples

The metabolic ratios lactate/pyruvate and β-hydroxybutyrate/acetoacetate are considered valuable tools to evaluate the in vivo redox cellular state by estimating the free NAD+/NADH in cytoplasm and mitochondria, respectively. The aim of the current study was to validate a gas-chromatography mass spectrometry method for simultaneous determination of the four metabolites in plasma and liver tissue. The procedure included an o-phenylenediamine microwave-assisted derivatization, followed by liquid-liquid extraction with ethyl acetate and silylation with bis(trimethylsilyl)trifluoroacetamide:trimethylchlorosilane 99:1. The calibration curves presented acceptable linearity, with a limit of quantification of 0.001 mM for pyruvate, β-hydroxybutyrate and acetoacetate and of 0.01 mM for lactate. The intra-day and inter-day accuracy and precision were within the European Medicines Agency’s Guideline specifications. No significant differences were observed in the slope coefficient of three-point standard metabolite-spiked curves in plasma or liver and water, and acceptable recoveries were obtained in the metabolite-spiked samples. Applicability of the method was tested in precision-cut liver rat slices and also in HepG2 cells incubated under different experimental conditions challenging the redox state. In conclusion, the validated method presented good sensitivity, specificity and reproducibility in the quantification of lactate/pyruvate and β-hydroxybutyrate/acetate metabolites and may be useful in the evaluation of in vivo redox states.     

A Novel DUF569 Gene Is a Positive Regulator of the Drought Stress Response in Arabidopsis

In the last two decades, global environmental change has increased abiotic stress on plants and severely affected crops. For example, drought stress is a serious abiotic stress that rapidly and substantially alters the morphological, physiological, and molecular responses of plants. In Arabidopsis, several drought-responsive genes have been identified; however, the underlying molecular mechanism of drought tolerance in plants remains largely unclear. Here, we report that the “domain of unknown function” novel gene DUF569 (AT1G69890) positively regulates drought stress in Arabidopsis. The Arabidopsis loss-of-function mutant atduf569 showed significant sensitivity to drought stress, i.e., severe wilting at the rosette-leaf stage after water was withheld for 3 days. Importantly, the mutant plant did not recover after rewatering, unlike wild-type (WT) plants. In addition, atduf569 plants showed significantly lower abscisic acid accumulation under optimal and drought-stress conditions, as well as significantly higher electrolyte leakage when compared with WT Col-0 plants. Spectrophotometric analyses also indicated a significantly lower accumulation of polyphenols, flavonoids, carotenoids, and chlorophylls in atduf569 mutant plants. Overall, our results suggest that novel DUF569 is a positive regulator of the response to drought in Arabidopsis.