A Novel Balanced-Lethal Host-Vector System Based on glmS

During the last decade, an increasing number of papers have described the use of various genera of bacteria, including E. coli and S. typhimurium, in the treatment of cancer. This is primarily due to the facts that not only are these bacteria capable of accumulating in the tumor mass, but they can also be engineered to deliver specific therapeutic proteins directly to the tumor site. However, a major obstacle exists in that bacteria because the plasmid carrying the therapeutic gene is not needed for bacterial survival, these plasmids are often lost from the bacteria. Here, we report the development of a balanced-lethal host-vector system based on deletion of the glmS gene in E. coli and S. typhimurium. This system takes advantage of the phenotype of the GlmS⁻ mutant, which undergoes lysis in animal systems that lack the nutrients required for proliferation of the mutant bacteria, D-glucosamine (GlcN) or N-acetyl-D-glucosamine (GlcNAc), components necessary for peptidoglycan synthesis. We demonstrate that plasmids carrying a glmS gene (GlmS⁺p) complemented the phenotype of the GlmS⁻ mutant, and that GlmS⁺p was maintained faithfully both in vitro and in an animal system in the absence of selection pressure. This was further verified by bioluminescent signals from GlmS ⁺pLux carried in bacteria that accumulated in grafted tumor tissue in a mouse model. The signal was up to several hundred-fold stronger than that from the control plasmid, pLux, due to faithful maintenance of the plasmid. We believe this system will allow to package a therapeutic gene onto an expression plasmid for bacterial delivery to the tumor site without subsequent loss of plasmid expression as well as to quantify bioluminescent bacteria using in vivo imaging by providing a direct correlation between photon flux and bacterial number.     

The Australian giant fleas Macropsylla Rothschild, 1905 (Siphonaptera: Macropsyllidae: Macropsyllinae), their identification,evolution, ecology,and conservation biology

Systematic Parasitology - Macropsylla Rothschild, 1905 is an endemic Australian flea genus represented by two species: M. hercules Rothschild, 1905 and M. novaehollandiae Hastriter, 2002. However,...     

Revisiting the use of Iprodione and Trichoderma in the integrated management of onion white rot

The biocontrol properties of Trichoderma species are well documented, but their effectiveness in antagonism of the problematic Sclerotium cepivorum, the causal agent of white rot in Allium species, appears limited with reports of significant control only relating to deliberately-mutated strains of Trichoderma. Our previous studies have indicated the possibility of using selected naturally-occurring strains of the antagonist in the suppression of other diseases; now in vitro and controlled environment in vivo studies have indicated that a degree of control of Onion White Rot is possible, and that the selected antagonist strains can be used in integrated treatments with Iprodione to good effect. The possible value of such treatments is considered in light of other approaches to the suppression of this continuing problem.     

“New Canadians are new conservatives”: race,incorporation and achieving electoral success in multicultural Canada

Despite declarations of race's irrelevance, the Conservative Party of Canada's (CPC) stance on race-related policies under former Prime Minister Stephen Harper's leadership was key to every electoral campaign it fought. Moreover, it is not despite but because of their race that racialized political elites have been incorporated into the CPC and its antecedents. Indeed, beginning in the 1990s, the inclusion of Asian Members of Parliament (MP) became for the Reform Party of Canada, a weapon in the struggle for electoral success; that is, part of an era of Conservative racial realignment. By tracing the role of Asian Conservative MP in the rise of the Reform Party since its 1987 inception and the electoral success of the CPC (2006–2015), this article explores how racialized political elites become crucial to the legitimization of the racial state when it is through a discourse of inclusion that exclusion is crafted.     

Synthesis and dual biological effects of hydroxycinnamoyl phenylalanyl/prolyl hydroxamic acid derivatives as tyrosinase inhibitor and antioxidant

We previously reported that caffeoyl-amino acidyl-hydroxamic acid (CA-Xaa-NHOH) acted as both a good antioxidant and tyrosinase inhibitor, in particular when caffeic acid was conjugated with proline or amino acids having aromatic ring like phenylalanine. Here, various hydroxycinnamic acid (HCA) derivatives were further conjugated with phenylalanyl hydroxamic acid and prolyl hydroxamic acid (HCA-Phe-NHOH and HCA-Pro-NHOH) to study the structure and activity relationship as both antioxidants and tyrosinase inhibitors. When their biological activities were evaluated, all HCA-Phe-NHOH and HCA-Pro-NHOH exhibited enhanced antioxidant activity compared to HCA alone. Moreover, derivatives of caffeic acid, ferulic acid, and sinapic acid inhibited lipid peroxidation more efficiently than vitamin E analogue (Trolox). In addition, derivatives of caffeic acid and sinapic acid efficiently inhibited tyrosinase activity and reduced melanin content in melanocytes Mel-Ab cell.     

Direct force measurement of single DNA–peptide interactions using atomic force microscopy

The selective interactions between DNA and miniature (39 residues) engineered peptide were directly measured at the single‐molecule level by using atomic force microscopy. This peptide (p007) contains an α‐helical recognition site similar to leucine zipper GCN4 and specifically recognizes the ATGAC sequence in the DNA with nanomolar affinity. The average rupture force was 42.1 pN, which is similar to the unbinding forces of the digoxigenin–antidigoxigenin complex, one of the strongest interactions in biological systems. The single linear fit of the rupture forces versus the logarithm of pulling rates showed a single energy barrier with a transition state located at 0.74 nm from the bound state. The smaller k_off compared with that of other similar systems was presumably due to the increased stability of the helical structure by putative folding residues in p007. This strong sequence‐specific DNA–peptide interaction has a potential to be utilized to prepare well‐defined mechanically stable DNA–protein hybrid nanostructures. Copyright © 2013 John Wiley & Sons, Ltd.     

Estimation of the ecosystem carbon budget in South Korea between 1999 and 2008

A carbon flux model, the vegetation integrated simulator for trace gases, was employed to estimate the carbon budgets of vegetation ecosystems in South Korea. The geographic information system was used to prepare the input variables for the model, such as climate, soil, and land-cover data, from reliable national inventories. Model simulation results indicated that the annual average gross primary production, net primary production, and soil respiration (SR) for 10 years were 91.89, 40.16, and 62.91 Tg C year^?1, respectively. The model also estimated a net ecosystem production with a value of 3.51 Tg C year^?1 between 1999 and 2008. Such results indicate that the vegetation ecosystems of South Korea offset 3.3 % of anthropogenic emissions as a net carbon sink. Latitudinal and topographical gradients over the total simulation area were found for all estimates. In addition, the estimates varied between seasons and years, especially in estimates for biomass growth and carbon uptake, because of variations in the weather conditions. Finally, model validation was conducted using measured soil efflux and flux measurement data from the Gwangneung experimental forest (GEF). The estimated SR accounted for 81.6 % of the observed SR at the GEF site (P < 0.005). Further, the model accounted well for the observed phase and amplitude of changes in the summer and autumn seasons.     

Enzymatic formation of carbohydrate rings catalyzed by single-walled carbon nanotubes

Macrocyclic carbohydrate rings were formed via enzymatic reactions around single-walled carbon nanotubes (SWNTs) as a catalyst. Cyclodextrin glucanotransferase, starch substrate and SWNTs were reacted in buffer solution to yield cyclodextrin (CD) rings wrapped around individual SWNTs. Atomic force microscopy showed the resulting complexes to be rings of 12–50 nm in diameter, which were highly soluble and dispersed in aqueous solution. They were further characterized by Raman and Fourier transform infrared spectroscopy and molecular simulation using density functional theory calculation. In the absence of SWNT, hydrogen bonding between glucose units determines the structure of maltose (the precursor of CD) and produces the curvature along the glucose chain. Wrapping SWNT along the short axis was preferred with curvature in the presence of SWNTs and with the hydrophobic interactions between the SWNTs and CD molecules. This synthetic approach may be useful for the functionalization of carbon nanotubes for development of nanostructures.