In vivo ligation of glucocorticoid-induced TNF receptor enhances the T-cell immunity to herpes simplex virus type 1

GITR (glucocorticoid-induced TNF receptor) is a recently identified member of the TNF receptor superfamily. The receptor is preferentially expressed on CD4(+)CD25(+) regulatory T cells and GITR signals break the suppressive activity of the subset. In this study, we wanted to reveal the in vivo function of GITR in herpes simplex virus type 1 (HSV-1) infection. A single injection of anti-GITR mAb (DTA-1) immediately after viral infection significantly increased the number of CD4(+) and CD8(+) T cells expressing CD25, an activation surface marker, and secreting IFN-gamma. We confirmed these in vivo observations by showing ex vivo that re-stimulation of CD4(+) or CD8(+) T cells with a CD4(+) or CD8(+) T-cell-specific HSV-1 peptide, respectively, induced a significant elevation in cell proliferation and in IFN-gamma secretion. Our results indicate that GITR signals play a critical role in the T-cell immunity to HSV-1.     

Ultraviolet photodissociation at 266 nm of phosphorylated peptide cations

Ultraviolet (UV) photodissociation (PD) experiments using 266 nm light were performed for a series of phosphopeptide cations in a Fourier transform mass spectrometer. The objective of the experiments was to determine whether 266 nm UV irradiation on the phosphopeptide cations would induce unique peptide backbone dissociation. In addition, the general behavior of the phosphate loss (?80 or ?98 Da) was monitored, particularly for those phosphopeptides with a phosphotyrosine residue that itself is a UV chromophore. For phosphopeptides with a UV chromophore, their photodissociation behavior was very similar to that of low‐energy sustained off‐resonance irradiation collisionally activated dissociation (SORI‐CAD), with a few exceptions. For example, b‐ and y‐type peptide backbone fragments were prevalent, and their dephosphorylation behavior was consistent with that of the SORI‐CAD results. For phosphoserine peptides, the loss of a phosphate group was always observed. On the other hand, for phosphotyrosine peptides, the phosphate loss was found to be dependent on the presence of a basic amino group in the sequence and the charge state of the precursor ions, in agreement with the CAD results in the literature. However, hydrogen atom loss or aromatic side chain loss, which is known to be the excited state specific fragmentation pathway, was rarely observed in our 266 nm UV PD experiments, in contrast to the previous UV PD literature (particularly at 220 nm). The mechanism for these observations is described in terms of dominant internal conversion followed by intramolecular vibrational energy redistribution (IVR). Copyright © 2009 John Wiley & Sons, Ltd.     

HepG2 Cell Resistance against Camptothecin from a Lysosomal Drug Delivery

A galactose‐appended drug delivery system released camptothecin (CPT) to lysosomes of HepG2 hepatoma cells, resulting in the cell resistance to the anticancer drug. We found that the resistance to CPT is caused by alteration of the drug release from the prodrug in lysosomes, emphasizing that the final delivery locations may critically influence drug efficacy.     

Development of various PS-b-P4VP micellar morphologies: fabrication of inorganic nanostructures from micellar templates

A liquid-phase method for preparing uniform-sized silica nanospheres (SNSs) 12 nm in size and their three-dimensionally ordered arrangement upon solvent evaporation have recently been pioneered by us. Here we report the successful control of the sphere sizes in the wide range from 14 to 550 nm by the seed regrowth method. In this method, the dispersion of SNSs 14 nm in size as seeds was prepared in the emulsion system containing Si(OEt)(4) (TEOS), water and arginine under weakly basic conditions (pH 9-10). An appropriate portion of this dispersion is added to the solution containing water, ethanol and arginine, and then TEOS is added. The additional TEOS introduced into the regrowth system contributed only to the resumed growth of the seeds, not to the formation of new silica particles. The size of interparticle pores was finely tuned by changing the size of the spheres. The preparation of three-dimensionally ordered porous carbons by using the colloidal array of silica nanospheres as a template is also reported.     

On the Support Size of Null Designs of Finite Ranked Posets

t -designs of the lattice of subspaces of a vector space over a finite field. The lower bound we find gives the tight bound for many important posets including the Boolean algebra, the lattice of subspaces of a vector space over a finite field, whereas the idea of the proofs of the main theorems makes it possible to prove that the lower bounds in the main theorems are not tight for some posets. Received: November 7, 1995     

Synthesis of Pure <Emphasis Type="Italic">meso</Emphasis>-2,3-Butanediol from Crude Glycerol Using an Engineered Metabolic Pathway in <Emphasis Type="Italic">Escherichia coli</Emphasis>

meso-2,3-Butanediol (meso-2,3-BDO) is essential for the synthesis of various economically valuable biosynthetic products; however, the production of meso-2,3-BDO from expensive carbon sources is an obstacle for industrial applications. In this study, genes involved in the synthesis of 2,3-BDO in Klebsiella pneumoniae were identified and used to genetically modify Escherichia coli for meso-2,3-BDO production. Two 2,3-BDO biosynthesis genes—budA, encoding acetolactate, and meso-budC, encoding meso-SADH—from K. pneumoniae were cloned into the pUC18 plasmid and introduced into E. coli. In 2 l batch culture, the SGSB03 E. coli strain yielded meso-2,3-BDO at 0.31 g/g_glucose (with a maximum of 15.7 g/l_culture after 48 h) and 0.21 g/g_{crude glycerol} (with a maximum of 6.9 g/l_culture after 48 h). Batch cultures were grown under optimized conditions (aerobic, 6% carbon source, 37 °C, and initial pH 7). To find the optimal culture conditions for meso-2,3-BDO production, we evaluated the enzyme activity of meso-SADH and the whole cell conversion yield (meso-2,3-BDO/acetoin) of the E. coli SGSB02, which contains pSB02. meso-SADH showed high enzyme activity at 30–37 °C and pH 7 (30.5–41.5 U/mg of protein), and the conversion yield of SGSB02 E. coli was highest at 37–42 °C and a pH of 7 (0.25–0.28 g _meso-2,3-BDO/g_acetoin).     

Salicylimine-based fluorescent chemosensor for aluminum ions and application to bioimaging

In this study, an assay to quantify the presence of aluminum ions using a salicylimine-based receptor was developed utilizing turn-on fluorescence enhancement. Upon treatment with aluminum ions, the fluorescence of the sensor was enhanced at 510 nm due to formation of a 1:1 complex between the chemosensor and the aluminum ions at room temperature. As the concentration of Al(3+) was increased, the fluorescence gradually increased. Other metal ions, such as Na(+), Ag(+), K(+), Ca(2+), Mg(2+), Hg(2+), Mn(2+), Co(2+), Ni(2+), Cu(2+), Zn(2+), Cd(2+), Pb(2+), Cr(3+), Fe(3+), and In(3+), had no such significant effect on the fluorescence. In addition, we show that the probe could be used to map intracellular Al(3+) distribution in live cells by confocal microscopy.