To determine the specificity of the microbial proteases in triggering the respiratory proteins to produce bactericidal ROS, we compared the level of bacterial clearance of protease-producing and non-producing strains of typical Gram-negative and Gram-positive bacteria. For Gram-negative bacteria, we chose Pseudomonas aeruginosa strain PAO-Iglewski which produces PAE (the major extracellular prote...

Authors: Naxin Jiang et al
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Polytriazoles have been shown recently to be useful ligands in the Cu(I)-catalyzed [3+2] cycloaddition of azides and alkynes.1 These ligands stabilize the Cu(I) species allowing efficient catalysis in aerobic, aqueous conditions. Since such ligands are prepared by the reaction they are used to facilitate, an autocatalytic effect is observed in their synthesis. Tristriazole 1 was prepared in the ma...

Authors: Holger B Kramer, Justin M Chalkner and Benjamin G Davis
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Fmoc determination Monitoring the resin loading during peptide synthesis gives useful information about the progress of the assembly. This can easily done by quantifying the amount of Fmoc removed at each deprotection/coupling cycle via spectrophotometry. Kaiser Test The Kaiser test1 is a qualitative test for the presence or absence of free primary amino groups, and it can be a useful indication a...

Authors: Irene Coin, Michael Beyermann and Michael Bienert
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Posttranslational histone modifications play important roles in regulating chromatin structure and function. One example of such modifications is histone ubiquitination, which occurs predominately on H2A and H2B. Although the recent identification of the ubiquitin ligase for histone H2A has revealed important roles for H2A ubiquitination in Hox gene silencing as well as in X inactivation, the enzy...

Authors: Heui-Yun Joo, Ling Zhai and Hengbin Wang
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In recent years, glycosyl thiols have become useful building blocks for the synthesis of certain glycoconjugates that may be considered to be analogues of glycopeptides and glycoproteins.1 Herein, we present a method that allows the direct conversion of reducing sugars to the corresponding glycoyl thiols. This method uses Lawesson’s reagent2 (LR) and was shown to be quite general and applicable ...

Authors: Gonçalo J. L. Bernardes and Benjamin G Davis
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Assays to detect cytokine production and degranulation are useful measures of T cell function. Multiparametric flow cytometry makes it possible to precisely phenotype and simultaneously detect these intermediate stages of T cell activation[1,2]. The degranulation of cytotoxic cells, an indicator of cytotoxic potential, results in the transient surface expression of CD107a and CD107b (lysosomal ass...

Authors: Joyce J Ruitenberg et al
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Crown-group crustaceans (Eucrustacea) are common in the fossil record of the past 500 million years back to the early Ordovician period, and very rare representatives are also known from the late Middle and Late Cambrian. With phosphatized cuticle and three-dimensional completion of soft parts, ‘Orsten’-type fossils, which are typically tiny embryos and cuticle-bearing animals, provide detaile...

Authors: Xi-guang Zhang et al
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In this protocol we described the isolation of retinal cells.

Authors: Cheng-Rong Yu et al
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This study implicates interleukin-17 (IL-17)-producing T-cells (TH17 cells) in etiology of human uveitis and scleritis. We show that blood of patients with uveitis or scleritis contained more TH17 cells than blood of healthy individuals. The numbers of TH17 cells increased during active uveitis/scleritis and decreased following treatment and in a mouse model, treatment with an IL-17-specific antib...

Authors: Charles Egwuagu et al
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The procedure consists of the following steps: Active induction of experimental autoimmune uveoretinitis (EAU) Preparation of antigen/CFA emulsion Immunization Clinical grading of mice with EAU by fundoscopy Preparation of sections for ocular histology Inhibition of EAU by IL-17 neutralizing antibody IL-27 expression in PBMC or retina is temporally correlated with progression of EAU

Authors: Charles Egwaugu et al
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