Preparation of a single-chain variable fragment and a recombinant antigen-binding fragment against the anti-malarial drugs, artemisinin and artesunate, and their application in an ELISA.

Two different recombinant antibodies, a single-chain variable fragment (scFv) and an antigen-binding fragment (Fab), were prepared against artemisinin (AM) and artesunate (AS) and were developed for use in an enzyme-linked immunosorbent assay (ELISA). The recombinant antibodies, which were derived from a single monoclonal antibody against AM and AS (mAb 1C1) prepared by us, were expressed by Escherichia coli cells and their reactivity and specificity were characterized. As a result, to obtain sufficient signal in indirect ELISA, a much greater amount of a first antibody was needed in the use of scFv due to the differences of the secondary antibody and conformational stability. Therefore, we focused on the development of the recombinant Fab antibodies and applied it to indirect competitive ELISA. The specificity of the Fab was similar to that of mAb 1C1 in that it showed specific reactivity toward AM and AS only. The sensitivity of the icELISA (0.16 µg/mL to 40 µg/mL for AM and 8.0 ng/mL to 60 ng/mL for AS) was sufficient for analysis of antimalarial drugs, and its utility for quality control of analysis of Artemisia spp. was validated. The Fab expression and refolding systems provided a good yield of high-quality antibodies. The recombinant antibody against AM and AS provides an essential component of an economically attractive immunoassay and will be useful in other immunochemical applications for the analysis and purification of antimalarial drugs.

3-(3-Phenoxybenzyl)amino-ß-carboline: a novel antitumor drug targeting α-tubulin.

3-(3-Phenoxybenzyl)amino-ß-carboline 2h showed extremely-high activity; the IC(50) value was 0.074 µM. To verify 2h-induced cell death types, we observed the chromatin condensation, the DNA fragmentation and activated caspase-3 using Hoechst 33342, agarose electrophoresis and western blot, and suggesting 2h-induced cell death type was apoptosis. Flow cytometry showed that 2h-treated cell was induced SubG1 cell population after G2/M cell cycle arrest. In addition, using affinity chromatography and peptide mass fingerprinting, we found that interacting protein with this compound was α-tubulin protein.

Human and environmental health challenges for the next decade (2010–2020).

The public health and environmental communities will face many challenges during the next decade. To identify significant issues that might be addressed as part of the International Life Sciences Institute (ILSI) Health and Environmental Sciences Institute (HESI) scientific portfolio, an expert group of key government, academic, and industry scientists from around the world were assembled in 2009 to map the current and future landscape of scientific and regulatory challenges. The value of the scientific mapping exercise was the development of a tool which HESI, individual companies, research institutions, government agencies, and regulatory authorities can use to anticipate key challenges, place them into context, and thus strategically refine and expand scientific project portfolios into the future.

ILSI Health and Environmental Sciences Institute (HESI), global leader in advancing translational science to create science-based solutions for a sustainable, healthier world.

The Health and Environmental Sciences Institute (HESI) is a non-profit scientific research organization based in Washington, D.C., U.S.A. HESI was established in 1989 as a global branch of the International Life Sciences Institute (ILSI) to provide an international forum to advance the understanding of scientific issues related to human health, toxicology, risk assessment and the environment. For the last 25 years, HESI has been the global leader to advance application of new science and technologies in the areas of human health, toxicology, risk assessment and environment. The core principle of "tripartite approach" and the multi-sector operational model have successfully supported HESI's scientific programs to create science-based solutions for a sustainable and healthier world. HESI's achievements include the dataset to guide the selection of appropriate supporting assays for carcinogenicity testing, a new testing framework for agricultural chemicals with enhanced efficacy, predictivity, and reduced animal usage, novel biomarkers of nephrotoxicity which provide data on the location of timing of drug effects in the kidney allowing for enhanced drug development, etc.