STP-A11, an oncoprotein of Herpesvirus saimiri augments both NF-kappaB and AP-1 transcription activity through TRAF6

Herpesvirus saimiri (HVS), a member of the gamma-herpesvirus family, encodes an oncoprotein called Saimiri Transforming Protein (STP) which is required for lymphoma induction in non-human primates. However, a detailed mechanism of STP-A11-induced oncogenesis has not been revealed yet. We first report that STP-A11 oncoprotein interacts with TNF-alpha receptor-associated factor (TRAF) 6 in vivo and in vitro. Mutagenesis analysis of the TRAF6-binding motif 10PQENDE15 in STP-A11 reveals that Glu (E)12 residue is critical for binding to TRAF6 and NF-kappaB activation. Interestingly, co-expression of E12A mutant, lack of TRAF6 binding, with cellular Src (Src) results in decreased transcriptional activity of Stat3 and AP-1, a novel target of STP-A11 compared to that of wild type. Furthermore, the presence of STP-A11 enhances the association of TRAF6 with Src and induces the translocation of both TRAF6 and Src to a nonionic detergent-insoluble fraction. Taken together, these studies suggest that STP-A11 oncoprotein up-regulates both NF-kappaB and AP-1 transcription activity through TRAF6, which would ultimately contribute cellular transformation.

Novel Vaccine Development against Escherichia coli O157: Immunologic Properties of O-Specific Polysaccharide-Shiga-like Toxin Conjugate Vaccines

Escherichia coli (E. coli) O157:H7 is an important cause of hemorrhagic colitis (HC) and hemolytic uremic syndrome (HUS). LPS-based vaccines need improvement since the anti-LPS antibodies raised by the vaccines are bactericidal and release toxin that may precipitate the development of HUS. Despite huge efforts, the treatment of infection with E. coli O157 has been difficult because antibiotics do not change the course of the enteritis caused by E. coli O157 and may increase the incidence of HUS through the release of Shiga-like toxin (Stx)-I. For this aim we tried the conjugate of E. coli O157 OSP bound to the nontoxic B subunit of Stx-I B as a vaccine that can induce both serum IgG anti-LPS antibody with bactericidal activity and neutralizing antibody to Stx-I. Mice were immunized s.c. with OSP-Stx-I B conjugate. Anti-sera were analyzed for the anti-LPS antibody, anti-Stx-I B antibody, complement-dependent bactericidal activity, and Stx-I neutralization activity. Mice injected with the bivalent conjugate elicited both bactericidal antibodies to E. coli O157 and neutralization antibodies to Stx-1. We also analyzed the distributional, functional changes of T lymphocytes in vitro and in vivo. After the injection of Stx-I, splenocytes showed a decrease in proliferation when stimulated with phytohemagglutinin (PHA) or LPS, and the number of CD4+ and CD8+ T cells also decreased. Interleukin (IL)-2, IL-4 and IFN-gamma productions by CD3+ T cells were slightly increased in the Stx-I injected mice.

Phylogenetic Analysis of Caterpillar Fungi by Comparing ITS 1-5.8S-ITS 2 Ribosomal DNA Sequences

This study was carried out to identify the phylogenetic relationships among several caterpillar fungi by comparing the sequences of internal transcribed spacer regions (ITS1 and ITS2) and 5.8S ribosomal DNA (rDNA) repeat unit. The sequences of ITS1, ITS2, and the 5.8S rDNA from 10 strains of Cordyceps species, 12 strains of Paecilomyces, 3 strains of Beauveria, 2 strains of Metarhizium and 1 strains of Hirsutella were amplified, determined and compared with the previously known Cordyceps species. The sequences of 5.8S rDNA were more conserved in length and variation than those of ITS regions. Although the variable ITS sequences were often ambiguously aligned, the conserved sites could be found. In the phylogenetic tree, the species generally divided into three clusters, supported by their morphology and/or host ranges. The 5.8S rDNA and ITS1 sequences among 10 species of Cordyceps militaris were identical and only one base pair in ITS2 sequence was different. Cordyceps sinensis and Cordyceps ophioglossoides were also clearly different, although they belonged to the same cluster. The GenBank database search of species revealed sister taxa of an entomogenous fungus. Metarhizium was used as an outgroup in all taxa.

Availability of Sikhae Factory Wastewater as a Submerged Culture Medium for Lentinula edodes

Sikhae is a Korean traditional beverage of saccharified rice. Its factory waste (SFW) is usually thrown away instead of being used. We developed a cheap substrate of SFW for use in liquid spawn that is known for its higher fruit body yields than grain spawn in sawdust cultivation. Mycelia of Lentinula edodes ASI 3046, which is regarded as the most suitable strain for sawdust cultivation, were cultured on six kinds of previous known media and SFW. As the seven kinds of media were applied, a Sikhae Factory Waste (SFW) was most excellent in growth. The dried mycelial weight in SFW was almost four times as much as that in the other media. In the flask culture, optimum culture conditions for the mycelial growth were obtained after 13 days of cultivation at media volume of 100 ml, 100 rpm, initial pH 4.5, and 25degrees C. The best mycelial growth was observed when MgSO4 . 7H2O and D-sucrose were added as a supplement in SFW. SWM must be a remarkable medium for L. edodes because of its simple preparation and low cost.