A day in the half-life of a protein

Researchers describe a method called bleach-chase to quantitatively measure the half-lives of fluorescently tagged proteins in human cancer cells.     

Effects of cycloSal-D4TMP Derivatives in H9 Cells with Induced AZT Resistance Phenotype

Abstract

Cytotoxic and antiretroviral activity of cycloSal-d4TMP derivatives were tested in a new AZT-resistant H9 cell subline (H9^rAZT²⁵⁰). The results showed, that cycloSal-d4TMP derivatives overcame resistance of HIV-1 to d4T in H9^rAZT²⁵⁰ cells, which exert decreased thymidine kinase (TK) gene expression.     

A quantum chemical investigation of the electronic structure of thionine

We have examined the electronic and molecular structure of 3,7-diaminophenothiazin-5-ium dye (thionine) in the electronic ground state and in the lowest excited states. The electronic structure was calculated using a combination of density functional theory and multi-reference configuration interaction (DFT/MRCI). Equilibrium geometries were optimized employing (time-dependent) density functional theory (B3LYP functional) combined with the TZVP basis set. Solvent effects were estimated using the COSMO model and micro-hydration with up to five explicit water molecules. Our calculated electronic energies are in good agreement with experimental data. We find the lowest excited singlet and triplet states at the ground state geometry to be of π→π* (S(1), S(2), T(1), T(2)) and n→π* (S(3), T(3)) character. This order changes when the molecular structure in the electronically excited states is relaxed. Geometry relaxation has almost no effect on the energy of the S(1) and T(1) states (~0.02 eV). The relaxation effects on the energies of S(2) and T(2) are moderate (0.14-0.20 eV). The very small emission energy results in a very low fluorescence rate. While we were not able to locate the energetic minimum of the S(3) state, we found a non-planar minimum for the T(3) state with an energy which is very close to the energy of the S(1) minimum in the gas phase (0.04 eV above). When hydration effects are taken into account, the n→π* states S(3) and T(3) are strongly blueshifted (0.33 and 0.46 eV), while the π→π* states are only slightly affected (<0.06 eV).     

Pushing size to the limit: extreme DNA

Double-wide pyrimidine bases can assemble into helical, sequence-specific DNA complexes.     

Proteins: caught in the trap

Two independent groups have recently devised innovative methods using light to trap and manipulate particles as small as proteins.     

SARS-coronavirus (SARS-CoV) and the safety of a solvent/detergent (S/D) treated immunoglobulin preparation.

SARS-coronavirus (SARS-CoV) is a newly emerged, highly pathogenic agent that caused over 8000 human infections with nearly 800 deaths between November 2002 and September 2003. While direct person-to-person transmission via respiratory droplets accounted for most cases, other modes have not been ruled out. SARS-CoV viraemia does not seem to reach high titres, however, it has to be excluded that virus transmission may occur via blood transfusion or application of therapeutic plasma products, e.g. fresh-frozen plasma or single components derived thereof. Manufacturing processes of all plasma derivatives are required to comprise dedicated virus inactivation/removal steps. Treatment with a mixture of solvent and detergent (SD) has successfully been applied to inactivate the most members of the transfusion-relevant viruses without affecting therapeutic properties of the products. The SD treatment irreversibly disrupts the lipid envelope of viruses such as HIV, HBV, HCV, HGV and CMV. In this study we evaluated the manufacturing process of an immunoglobulin preparation (OCTAGAM, manufactured by Octapharma Pharmazeutika Produktionsges.m.b.H., Vienna, Austria) for its capacity to inactivate the SARS-CoV. Our results demonstrate that SARS-CoV was completely inactivated below the limit of detection. This was found to occur within 1 min of SD treatment.     

Effect of pesticides on Colpoclypeus florus (Hymenoptera: Eulophidae) and Trichogramma platneri (Hymenoptera: Trichogrammatidae), parasitoids of leafrollers in Washington.

Pesticides were evaluated for their effect on two parasitoid species, Colpoclypeus flouts and Trichogramma platneri, that are potential biological control agents of leafrollers in apple orchards. Organophosphate and carbamate insecticides were highly toxic to both parasitoids in topical applications, but foliar residues of some products were nontoxic after 7 d. At reduced rates, topically applied pyrethroids were low in toxicity to C. florus were highly toxic to T. platneri, and foliar residues were nontoxic after about 7 d. Imidacloprid and abamectin were highly toxic when applied topically to both parasitoids but were not toxic as 1-d-old residues. Insect growth regulators did not cause mortality either as topical applications or residues; however, diflubenzuron caused severe sublethal effects, completely blocking the production of C. florus offspring. Biorational pesticides, such as soap, oil, and B. thuringiensis products, caused no toxicity to C. florus but had a direct impact on T. platneri as topical applications through physical immobilization. The potential to integrate different pesticides with biological control of leafrollers and the need for a step-wise approach to evaluate the impact of pesticides against natural enemies is discussed.