Cytotoxicity, haematotoxicity and genotoxicity of high molecular mass arborescent polyoxyethylene polymers with polyglycidol-block-containing shells

We have examined the impact on biological systems of some newly synthesised polyoxyethylene polymers using in vitro assays. Toxicity was tested by the 3-[4,5-dimethylthiazole-2-yl]-2,5 diphenyltetrazolium bromide (MTT) assay, haemolysis was assessed, and an ethidium bromide (EB) assay was used to study interactions between the polymers and DNA. All the assay data showed that the polymers are biocompatible. No differences were found between generations (i.e. macromolecule sizes). The results encourage continuing studies on the clinical use of these molecules as drug carriers.     

Structural studies and biological evaluation of T30695 variants modified with single chiral glycerol-T reveal the importance of LEDGF/p75 for the aptamer anti-HIV-integrase activities

Some G-quadruplex (GQ) forming aptamers, such as T30695, exhibit particularly promising properties among the potential anti-HIV drugs. T30695?G-quadruplex binds to HIV-1 integrase (IN) and inhibits its activity during 3′-end processing at nanomolar concentrations. Herein we report a study concerning six T30695-GQ variants, in which the R or S chiral glycerol T, singly replaced the thymine residues at the T30695?G-quadruplex loops. CD melting, EMSA and HMRS experiments provided information about the thermal stability and the stoichiometry of T30695-GQ variants, whereas CD and ¹H NMR studies were performed to evaluate the effects of the modifications on T30695-GQ topology. Furthermore, LEDGF/p75 dependent and independent integration assays were carried out to evaluate how T loop modifications impact T30695-GQ biological activities. The obtained results showed that LEDGF/p75 adversely affects the potencies of T30695 and its variants. The IN inhibitory activities of the modified aptamers also depended on the position and on the chirality (R or S) of glycerol T loop in the GQ, mostly regardless of the G-quadruplex stabilities. In view of our and literature data, we suggest that the allosteric modulation of IN tetramer conformations by LEDGF/p75 alters the interactions between the aptamers and the enzyme. Therefore, the new T30695 variants could be suitable tools in studies aimed to clarify the HIV-1 IN tetramers allostery and its role in the integration activity.     

Hemolysis caused by polyoxyethylene-derived surfactants evidence for peroxide participation

The hemolytic properties of nonionic surfactants of the series $\text{CH}{}_3\text{(}\text{CH}{}_2\text{)}{}_{\mathrm{15–17}}\text{-}\text{O}\text{-(}\text{CH}{}_2\text{CH}{}_2\text{O}\text{)}{}_{\mathrm{n}}\text{CH}{}_2\text{CH}{}_2\text{OH}$ were investigated and compared to those of saponins, sapogenins and H₂O₂. Antioxidants and anaerobic conditions were shown to inhibit the hemolysis, while glycyrrhizin was found to enhance it. Similar effects were obtained for H₂O₂ hemolysis, but not for saponin and sapogenin hemolysis. It is proposed that peroxides and free radicals are mainly responsible for the polyoxyethylene derived surfactants induced hemolysis.     

Application of hydrophilic–lipophilic balance (HLB) number to optimize a compatible non-ionic surfactant for dried aerial conidia of Beauveria bassiana

The hydrophilic–lipophilic balance (HLB) number system was used to optimize a compatible non-ionic surfactant, TDA (polyoxyethylene tridecyl ether) in formulations for two Beauveria bassiana strains, NI8 and GHA. The optimal HLB number for TDA was determined on the basis of wetting times for conidial powders. The results indicated that optimal HLB number of TDA for B. bassiana strain NI8 was 8, while the optimum HLB number for strain GHA was 10. The optimized TDA surfactants required significantly less wetting times than the commonly used laboratory surfactants, Triton X-100, Span 80, and Tween 80. These optimized TDA surfactants were further characterized on their ability to produce conidial suspensions of the two strains after 5 min of mixing, TDA HLB 8 and TDA HLB 10 produced suspensions of 1.8 × 10⁸ and 1.6 × 10⁸ conidia/ml for NI8 and GHA, respectively. These conidial levels were significantly higher than those in Triton X-100, Span 80, and Tween 80 suspensions after the same mixing time. Germination assays showed that TDA HLB 8 promoted significantly higher germination rates of strain NI8 than those observed in other commonly used laboratory surfactants. However, the germination rates of the GHA strain were unaffected by any of the surfactants tested. The efficacy of the conidial suspensions was confirmed with assays against Lygus lineolaris. Bioassay results indicated that there were no significant differences in mortalities because of surfactants. These results suggest optimization based upon HLB number will not negatively impact parameters associated with efficacy, while providing desirable physical properties.     

Detergent extraction of membrane proteins related to the action of antidiuretic hormone

Antidiuretic hormone (ADH) induces, in the apical plasma membrane of target cells, the insertion of intramembranous particle aggregates that probably contain water channels. A mild attack of this membrane by a polyoxyethylene nonylphenyl detergent, which reversibly depressed ADH-induced water permeability, has been found to modify aggregate structure while extracting additional proteins. This simple procedure could be a valuable approach to the problem of aggregate isolation and characterization.