Human Brain Astrocytes Mediate TRAIL-mediated Apoptosis after Treatment with IFN-γ

TNF-related apoptosis inducing ligand (TRAIL) expressions were studied in primary human brain astrocytes in response to pro-inflammatory cytokines. When astrocytes were treated with IL-1β, TNF-α or IFN-γ, TRAIL was induced in cultured fetal astrocytes. In particular, IFN-γ induced the highest levels of TRAIL in cultured astrocytes. When astrocytes were prereated with IFN-γ, they induced apoptosis in TRAIL-sensitive Peer cells. Our results suggest that IFN-γ modulates the expression of TRAIL in astrocytes, which may enhance cytotoxic sensitivity of infiltrating immune cells or brain cells other than astrocytes during inflammation of brain

Two novel mutations of Wiskott–Aldrich syndrome: the molecular prediction of interaction between the mutated WASP L101P with WASP-interacting protein by molecular modeling

AbstractWiskott–Aldrich syndrome (WAS) is an X-linked disorder characterized by eczema, thrombocytopenia and increased susceptibility of infections, with mutations of the WAS gene being responsible for WAS and X-linked thrombocytopenia. Herein, two novel mutations of WAS at T336C on exon 3, and at 1326–1329, a G deletion on exon 10, resulting in L101P missense mutation and frameshift mutation 444 stop, respectively, are reported. The affected patients with either mutation showed severe suppression of WAS protein (WASP) levels, T cell proliferation, and CFSE-labeled T cells division. Because WASP L101 have not shown direct nuclear Overhauser effect (NOE) contact with the WASP-interacting protein (WIP) in NMR spectroscopy, molecular modeling was performed to evaluate the molecular effect of WASP P101 to WIP peptide. It is presumed that P101 induced a conformational change in the Q99 residue of WASP and made the side chain of Q99 move away from the WIP peptide, resulting in disruption of the hydrogen bond between Q99 WASP and Y475 WIP. A possible model for the molecular pathogenesis of WAS has been proposed by analyzing the interactions of WASP and WIP using a molecular modeling study

Development of a high yield purification process for the production of influenza virus vaccines

Production of influenza virus in animal cells has emerged as an alternative to conventional platforms such as egg-based production system. Animal cells, especially MDCK and VERO cell lines, are widely used as the primary production cell for influenza virus vaccine because of their high susceptibility to infection with various influenza viruses. Recently, a robust and reliable purification process was successfully developed for the production of quadri-valent HA proteins (from two strains of the type A virus and two strains of the type B virus) by using animal cell-based production system in Green Cross Corp., Korea. The UF/DF process, Benzonase treatment at high temperature as well as column chromatography strategy was optimized to maximize the final HA production yields. Benzonase treatment was conducted to reduce in hcDNA (host cell DNA) because hcDNA was main impurity for cell-based influenza virus vaccine. A simple and stable UF/DF process has been tested with membrane molecular weight cutoffs of 100 and 300 kDa as well as 0.2 and 0.45 um microfiltration membrane. Anion exchange chromatography (AEC) and size exclusion chromatography (SEC) were selected for acceptable reduction in hcDNA and HCP. AEC was used to separate hcDNA from virus at a salt concentration of 0.5 M sodium chloride. The HA yield through AEC & SEC combination process was sufficiently achieved under specific purification process condition. Overall, the amount of residual hcDNA was reduced to an acceptable level (10ng/dose) and the increased HA yield was maintained throughout the whole process. The performance, productivity and scalability of the purification process were successfully demonstrated in over 30 GMP batches using 4 different influenza virus strains

The Computerized Design Program for Tunnel Blasting

Abstract In this study, a computer program to design tunnel blasting pattern has been developed. The program consists of two parts; one is for tunnel blasting pattern design and the other is for blasting modeling to estimate the peak particle velocity, the distribution of fragmentation and the excavation damage zone. We modified the design method of tunnel blasting pattern suggested by Langefors because it provided undesirable pattern in blasting practices such as considerably large center cut and too large burden for Vcut as drilling length increased. As a result, the burden and spacing were reduced to practically appropriate amounts. In addition, the correlation between rock mass rate, RMR, and rock constant in blasting, c, was analyzed based on the data collected from twenty three tests of tunnel blasting. It was concluded that the correlation between them was fairly good enough to be applied in cut design. In order to check the validity of the modified methods and their practical applicability, test blasting was carried out at two different tunnel construction sites in Korea. The results were satisfactory in that the average rate of advance was 90% and the overbreak did not cause additional support. Futhermore, the developed program is capable of estimating peak particle velocity by using (a) the existing vibration equations, (b) the vibration equation obtained by test blasting to check out the practical applicability of the designed blasting pattern. Feedback is implemented into the program to adjust the designed blasting pattern and control the vibration

Edge-functionalized graphene-like platelets as a co-curing agent and a nanoscale additive to epoxy resin

A newly developed method for the edge-selective functionalization of "pristine" graphite with 4-aminobenzoic acid was applied for the synthesis of 4-aminobenzoyl-functionalized graphite (AB-graphite) through a "direct" Friedel-Crafts acylation in a polyphosphoric acid (PPA)/phosphorus pentoxide medium (P(2)O(5)). The AB moiety at the edge of the AB-graphite played the role of a molecular wedge to exfoliate the AB-graphite into individual graphene and graphene-like platelets upon dispersion in polar solvents. These were used as a co-curing agent and a nanoscale additive to epoxy resin. The physical properties of the resulting epoxy/AB-graphite composites were improved because of the efficient load transfer between the additive and epoxy matrix through covalent links.close191

Development of Monoclonal Antibodies Against Human IRF-5 and Their Use in Identifying the Binding of IRF-5 to Nuclear Import Proteins Karyopherin-α1 and -β1

PURPOSE: IRF-5 is a direct transducer of virus-mediated and TLR-mediated signaling pathways for the expression of cytokines and chemokines which form homodimers or heterodimers with IRF-7. However, direct IRF-5-specific monoclonal antibodies (mAbs) are not available at present. These could be used to further evaluate the functions of IRF-5. In this study, we produced and characterized three mouse mAbs to human IRF-5. The binding of IRF-5 to nuclear import proteins was first identified using a mAb. MATERIALS AND METHODS: His-tagged human IRF-5 protein spanning amino acid residues 193-257 was used as an antigen and three mAbs were produced. The mAbs were tested with ELISA, Western blot analysis (WB), immunofluorescent staining (IF), and immunoprecipitation (IP). In addition, the nuclear import protein which carried phosphorylated IRF-5 was identified using one of these mAbs. RESULTS: MAbs 5IRF8, 5IRF10 and 5IRF24 which reacted with the recombinant His-IRF-5(193-257) protein were produced. All mAbs bound to human IRF-5, but not to IRF-3 or IRF-7. They could be used for WB, IF, and IP studies. The binding of phosphorylated IRF-5 to karyopherin-alpha1 and -beta1 was also identified. CONCLUSION: Human IRF-5-specific mAbs are produced for studying the immunologic roles related to IRF-5. Phosphorylated IRF-5 is transported to the nucleus by binding to nuclear import proteins karyopherin-alpha1 and -beta1.ope

BOAO Photometric Survey of Galactic Open Clusters. III. Czernik 24 and Czernik 27

We present BV CCD photometry for the open clusters Czernik 24 and Czernik 27. These clusters have never been studied before, and we provide, for the first time, the cluster parameters; reddening, distance, metallicity and age. Czernik 24 is an old open cluster with age 1.8 +/- 0.2 Gyr, metallicity [Fe/H]=-0.41 +/- 0.15 dex, distance modulus (m-M)_0 = 13.1 +/- 0.3 mag (d=4.1 +/- 0.5 kpc), and reddening E(B-V) = 0.54 +/- 0.12 mag. The parameters for Czernik 27 are estimated to be age = 0.63 +/- 0.07 Gyr, [Fe/H]= -0.02 +/- 0.10 dex, (m-M)_0 = 13.8 +/- 0.2 mag (d=5.8 +/- 0.5 kpc), and E(B-V) = 0.15 +/- 0.05 mag. The metallicity and distance values for Czernik 24 are consistent with the relation between the metallicity and the Galactocentric distance of other old open clusters. We find the metallicity gradient of 51 old open clusters including Czernik 24 to be Delta [Fe/H]/Delta R_gc= -0.064 +/- 0.009 dex/kpc.Comment: Accepted by the Journal of the Korean Astronomical Society, 2005 December issu

Anti-inflammatory effects of Radix Gentianae Macrophyllae (Qinjiao), Rhizoma Coptidis (Huanglian) and Citri Unshiu Pericarpium (Wenzhou migan) in animal models

<p>Abstract</p> <p>Background</p> <p>KHU14, an ethanolic extract of <it>Radix Gentianae Macrophyllae </it>(<it>Qinjiao</it>), <it>Rhizoma Coptidis </it>(<it>Huanglian</it>) and <it>Citri Unshiu Pericarpium </it>(<it>Wenzhou migan</it>) was tested for its anti-inflammatory effects.</p> <p>Methods</p> <p>Three out of 20 herbs were found to have anti-inflammatory effects. The formulation of these herbs, i.e. KHU14 was tested for croton oil-induced ear edema, carrageenan-induced paw edema, acetic acid-induced capillary permeability, cotton pellet and delayed type hypersensitivity.</p> <p>Results</p> <p>KHU14 exhibited anti-inflammatory effects in animal models of acute and chronic inflammation. The anti-inflammatory activity of KHU14 observed was comparable to that of celecoxib. KHU14 inhibited the production of NO and PGE₂in LPS/IFN-gamma-stimulated peritoneal macrophages, and reduced edema and the amount of infiltrated cells in animal models.</p> <p>Conclusion</p> <p>KHU14 exhibited anti-inflammatory effects as demonstrated in typical immunological tests for anti-inflammation <it>in vitro </it>and <it>in vivo</it>.</p