Splenic CD8(+) T cells secrete TGF-beta 1 to exert suppression in mice with anterior chamber-associated immune deviation

Background CD8(+) regulatory T cells (Treg) have been considered to be involved in a model of ocular-induced tolerance, known as anterior chamber-associated immune deviation (ACAID). The mechanisms of suppression by CD8(+) T cells in ACAID remain only poorly understood. TGF-beta 1 is considered as an inhibitory cytokine for immunosuppression in some models. The production of TGF-beta 1 by CD8(+) T cells in ACAID, and whether CD8+ T cells exert suppression through TGF-beta 1, is unknown. Methods The suppressive effect of CD8(+) T cells in ACAID mice was determined by a local adoptive transfer (LAT) assay. The production of TGF-beta 1 by CD8(+) T cells was measured by enzyme-linked immunosorbent assay (ELISA). Anti-TGF-beta 1 antibodies were used in the LAT assay to test if they could block the inhibitory effect of CD8(+) T cells. Results CD8(+) T cells from ACAID mice were shown to block the delayed-type hypersensitivity (DTH) response in an antigen-specific manner in a LAT assay. These CD8+ T cells secreted TGF-beta 1, and their suppression could partially be blocked by anti-TGF-beta 1 antibodies. Conclusions Our study confirms that CD8+ T cells from ACAID mice possess inhibitory properties. This population exerts part of its suppressive function via the production of TGF-beta 1

Retinal S-antigen Th1 cell epitope mapping in patients with Behcet's disease

Background - Retinal S-antigen (S-Ag) is a most characterized autoantigen of autoimmune uveitis. The recognized immunodominant epitope of human S-Ag in patients with uveitis has not been identified. In this study, we selected certain patients with active uveitis to map the Th1 cell epitope spectrum of human S-Ag in Behcet's disease(BD). Methods - Blood samples were taken from eight active BD patients who showed an immune response to 40 mixed overlapping peptides spanning the entire sequence of human S-Ag. Peripheral blood mononuclear cells were isolated and stimulated with single S-Ag peptide at 5 mu g/ml or 20 mu g/ml. Single-cell immune responses were measured by IFN-gamma ELIspot assay. Results - BD patients heterogeneously responded to the S-Ag peptides at two concentrations. In general, the responses to 5 mu g/ml peptides were slightly stronger than those to 20 mu g/ml peptides, while the maximum SFC frequency to single peptide at the two concentrations was similar. Several peptides including P31, P35 and P40 induced a prominent response, with the frequency of S-Ag specific cells being about 0.007%. Significant reactivity pattern shift was noted in patients with different disease courses. Conclusions - Certain active BD patients have S-Ag specific Th1 cells with a low frequency. The S-Ag epitope specificity between patients is highly heterogeneous, and varies with the uveitis cours

Epitope recognition of peptide-imprinted polymers for Regenerating protein 1 (REG1)

Molecularly imprinted polymers (MIPs) were developed to replace natural antibodies with a cost-effective and durable synthetic material. Molecular imprinting of proteins conventionally utilizes the whole protein as the template, which is complex (as many different epitopes may be imprinted) and expensive. In this work, seven peptides (13–18 amino acids) were synthesized and used as templates for the imprinting and recognition of Regenerating Protein 1 (REG1). REG1 is involved in the proliferation and differentiation of diverse cell types, and was recently described as a urinary biomarker for pancreatic ductal adenocarcinoma (PDAC). Peptide-imprinted poly(ethylene-co-vinyl alcohol)s (PIPs), containing four different mole fractions of ethylene were cast on screen-printed electrodes to find the optimum composition for both the sensing and the extraction of REG1 in an E. coli culture medium. Peptides with fewer than 16 amino acids and two or three aromatic and hydrophobic groups have a higher affinity for MIPs of poly(ethylene-co-vinyl alcohol) (EVAL) with 27 mol% of ethylene, while those with four aromatic and hydrophobic groups have a higher affinity for MIPs with EVALs that contain 32 mol% of ethylene. The peptide / EVAL combination that maximized both imprinting effectiveness and response to REG1B was the sequence NEDRETWVDADLY imprinted into 32 mol% EVAL. This EVAL composition and template peptide were then modified by incorporation of magnetic nanoparticles, thus extending applications for PIPs to include extraction of REG1 protein from E. coli culture medium

Hamiltonian Formalism of the de-Sitter Invariant Special Relativity

Lagrangian of the Einstein's special relativity with universal parameter $c$ ($\mathcal{SR}_c$) is invariant under Poincar\'e transformation which preserves Lorentz metric $\eta_{\mu\nu}$. The $\mathcal{SR}_c$ has been extended to be one which is invariant under de Sitter transformation that preserves so called Beltrami metric $B_{\mu\nu}$. There are two universal parameters $c$ and $R$ in this Special Relativity (denote it as $\mathcal{SR}_{cR}$). The Lagrangian-Hamiltonian formulism of $\mathcal{SR}_{cR}$ is formulated in this paper. The canonic energy, canonic momenta, and 10 Noether charges corresponding to the space-time's de Sitter symmetry are derived. The canonical quantization of the mechanics for $\mathcal{SR}_{cR}$-free particle is performed. The physics related to it is discussed.Comment: 24 pages, no figur

Phylogenetic analysis and biochemical characterization of a thermostable dihydropyrimidinase from alkaliphilic Bacillus sp TS-23

Two degenerate primers established from the alignment of highly conserved amino acid sequences of bacterial dihydropyrimidinases (DHPs) were used to amplify a 330-bp gene fragment from the genomic DNA of Bacillus sp. TS-23 and the amplified DNA was successfully used as a probe to clone a dhp gene from the strain. The open reading frame of the gene consisted of 1422 bp and was deduced to contain 472 amino acids with a molecular mass of 52 kDa. The deduced amino acid sequence exhibited greater than 45% identity with that of prokaryotic D-hydantoinases and eukaryotic DHPs. Phylogenetic analysis showed that Bacillus sp. TS-23 DHP is grouped together with Bacillus stearothermophilus D-hydantoinase and related to dihydroorotases and allantoinases from various organisms. His(6)-tagged DHP was over-expressed in Escherichia coli and purified by immobilized metal affinity chromatography to a specific activity of 3.46 U mg(-1) protein. The optimal pH and temperature for the purified enzyme were 8.0 and 60 degrees C, respectively. The half-life of His(6)-tagged DHP was 25 days at 50 degrees C. The enzyme activity was stimulated by Co2+ and Mn2+ ions. His(6)-tagged DHP was most active toward dihydrouracil followed by hydantoin derivatives. The catalytic efficiencies (k(cat)/K-m) of the enzyme for dihydrouracil and hydantoin were 2.58 and 0.61 s(-1) mM(-1), respectively

Synthesis and trans-ureation of N,N '-diphenyl-4, 4 '-methylenediphenylene biscarbamate with diamines: a non-isocyanate route (NIR) to polyureas

A non-isocyanate route (NIR) of making polyureas of high molecular weight has been found through transureation of N,N'-diphenyl-4,4'-methylenediphenylene biscarbamate (4,4'-DP-MDC) with a variety of diamines and mixed diamines. The preparation of 4,4'-DP-MDC was achieved readily by carbonylation of 4,4'-methylenedianiline (4,4'-MDA) with diphenyl carbonate (DPC) using organic acids as catalysts. It was found that the highest yield (99%) of pure 4,4'-DP-MDC can be isolated in a toluene solution under mild conditions co-catalyzed by benzoic acid and tertiary amine. Trans-ureation of 4,4'-DP-MDC with aliphatic amines indicated that the process is a highly solvent dependent process and was found to be extremely facile in dimethyl sulfoxide (DMSO) at 80 C and in tetramethylene sulfone (TMS) at 140 C in absence of any catalyst. Particularly, the most effective polymerization process was developed using tetramethylene sulfone (TMS) as the solvent under reduced pressure for concurrently distilling off phenol from the reaction mixture during the polymerization in a shifting equilibrium towards polyurea. However, this solvent-assisted transureation was found to be in-efficient when N, N'-dimethyl-4,4'- methylenediphenylene biscarbamate (4,4'-DM-MDC) was used in a similar condition for comparison. Thus, an efficient green-chemistry process has been developed based on 4,4'-DP-MDC in making urea prepolymers, urea elastomers and urea plastics all in excellent yields without using reactive methylenediphenylene diisocyanate (MDI) or any catalysts in the trans-ureation polymerizations

Characterization study of GaN-based epitaxial layer and light-emitting diode on nature-patterned sapphire substrate

[[abstract]]Chemical wet etching on c-plane sapphire wafers by three etching solutions (H3PO4, H2SO4, and H3PO4/H2SO4 mixing solution) was studied. Among these etching agents, the mixing H3PO4/H2SO4 solution has the fastest etching rate (1.5 μm/min). Interestingly, we found that H2SO4 does not etch the c-plane sapphire wafer in thickness; instead, a facet pyramidal pattern is formed on the c-plane sapphire wafer. GaN light-emitting diode (LED) epitaxial structure was grown on the sapphire wafer with the pyramidal pattern and the standard flat sapphire wafer. X-ray diffraction and photoluminescence measurement show that the pyramidal pattern on the sapphire wafer improved crystalline quality but augmented the compressive stress level in the GaN LED epilayer. The horizontal LED chips fabricated on the pyramidal-patterned sapphire wafer have a larger light output than the horizontal LED chips fabricated on the standard flat sapphire wafer by 20%.[[notice]]補正完畢[[incitationindex]]SCI[[booktype]]紙本[[booktype]]電子

Comparison of Genomes of Three Xanthomonas oryzae Bacteriophages

<p>Abstract</p> <p>Background</p> <p>Xp10 and OP1 are phages of <it>Xanthomonas oryzae </it>pv. oryzae (Xoo), the causative agent of bacterial leaf blight in rice plants, which were isolated in 1967 in Taiwan and in 1954 in Japan, respectively. We recently isolated the Xoo phage Xop411.</p> <p>Results</p> <p>The linear Xop411 genome (44,520 bp, 58 ORFs) sequenced here is 147 bp longer than that of Xp10 (60 ORFs) and 735 bp longer than that of OP1 (59 ORFs). The G+C contents of OP1 (51%) and Xop411 and Xp10 (52% each) are less than that of the host (65%). The 9-bp 3'-overhangs (5'-GGACAGTCT-3') in Xop411 and Xp10 are absent from OP1. More of the deduced Xop411 proteins share higher degrees of identity with Xp10 than with OP1 proteins, while the right end of the genomes of Xp10 and OP1, containing all predicted promoters, share stronger homology. Xop411, Xp10, and OP1 contain 8, 7, and 6 freestanding HNH endonuclease genes, respectively. These genes can be classified into five groups depending on their possession of the HNH domain (HNN or HNH type) and/or AP2 domain in intact or truncated forms. While the HNN-AP2 type endonuclease genes dispersed in the genome, the HNH type endonuclease genes, each with a unique copy, were located within the same genome context. Mass spectrometry and N-terminal sequencing showed nine Xop411 coat proteins, among which three were identified, six were assigned as coat proteins (4) and conserved phage proteins (2) in Xp10. The major coat protein, in which only the N-terminal methionine is removed, appears to exist in oligomeric forms containing 2 to 6 subunits. The three phages exhibit different patterns of domain duplication in the N-terminus of the tail fiber, which are involved in determination of the host range. Many short repeated sequences are present in and around the duplicated domains.</p> <p>Conclusion</p> <p>Geographical separation may have confined lateral gene transfer among the Xoo phages. The HNN-AP2 type endonucleases were more likely to transfer their genes randomly in the genome and may degenerate after successful transmission. Some repeated sequences may be involved in duplication/loss of the domains in the tail fiber genes.</p

Cyclophosphamide induces NR2B phosphorylation-dependent facilitation on spinal reflex potentiation

Chang CH, Peng HY, Wu HC, Lai CY, Hsieh MC, Lin TB. Cyclophosphamide induces NR2B phosphorylation-dependent facilitation on spinal reflex potentiation. Am J Physiol Renal Physiol 300: F692-F699, 2011. First published November 24, 2010; doi:10.1152/ajprenal.00531.2010.-It is well-established that cyclophosphamide (CYP) can sensitize the pelvic afferent nerve arising from the urinary bladder and therefore induce suprapubic pain. To test the possibility that CYP might mediate the development of visceral hypereflexia/hyperalgesia by facilitating spinal activity-dependent neural plasticity, we compared the pelvic-urethra reflex activity and spinal N-methyl-D-aspartate receptor NR2B subunit (NR2B) phosphorylation in rats treated with vehicle solution and CYP. Compared with vehicle solution, when accompanied by upregulation of phosphorylated NR2B expression in the lumbosacral (L6-S2) dorsal horn, CYP increased the evoked spikes in spinal reflex potentiation induced by repetitive stimulation (1 stimulation/1 s). Moreover, intraperitoneal pretreatments with N(G)-nitro-L-arginine methyl ester and roscovitine, nitric oxide synthase and cyclin-dependent protein kinase 5 (Cdk5) antagonists, respectively, overwrote CYP-enhanced reflex potentiation and NR2B phosphorylation. When compared with the untreated group, the treatment with small-interfering RNA of NR2B, which decreased the expression of NR2B expression, abolished CYP-dependent reflex facilitation and spinal NR2B phosphorylation. These results suggested that CYP might facilitate spinal reflex potentiation mediated by N-methyl-D-aspartate receptors and participate in the development of visceral hypereflexia/hyperalgesia through nitric oxide-and Cdk5-dependent NR2B phosphorylation at the lumbosacral dorsal horn