Influence of DNA repair by ada and ogt alkyltransferases on the mutational specificity of alkylating agents

We investigated the influence of the alkyltransferases (ATases) encoded by the ada and ogt genes of Escherichia coli on the mutational specificity of alkylating agents. A new mutational assay for selection of supF- mutations in shuttle-vector plasmids was used. Treating plasmid-bearing bacteria with N-methyl-N-nitrosourea (MNU), N-ethyl-N-nitrosourea (ENU), and ethyl methanesulfonate (EMS) dramatically increased the mutation frequency (from 33-fold to 789-fold). The vast majority of mutations (89-100%) were G:C-->A:T transitions. This type of mutation increased in ada- (MNU) or ogt- (ENU) bacteria, suggesting that repair of O6-methylguanine by ada ATase and repair of O6-ethylguanine by ogt ATase contribute mainly to the decrease in G:C-->A:T transitions. The analysis of neighboring base sequences revealed an overabundance of G:C-->A:T transitions at 5'-GG sequences. The 5'-PuG bias increased in ATase-defective cells, suggesting that these sequences were not refractory to repair. G:C-->A:T transitions occurred preferentially in the untranscribed strand after in vivo exposure. That this strand specificity was detected even in bacteria devoid of ATase activity (ada- ogt-) and not after in vitro mutagenesis suggests a bias for damage induction rather than for DNA repair. Highly significant differences were found between the in vivo and in vitro incidences of G:C-->A:T substitutions at the two major hotspots, positions 123 (5'-GGG-3'; antisense strand) and 168 (5'-GGA-3'; sense strand). These results are explained by differences in the probability of formation of stem-loop structures in vivo and in vitro.     

LSTSQ: a module for reliable constrained and unconstrained nonlinear regression

A software module for nonlinear regression analysis, based on the reliable Meyer-Roth algorithm (a modified damped least square algorithm), is presented. It allows both constrained and unconstrained optimization, and the use of a variety of weighting methods. Virtually any nonlinear function can be fitted, including those with several nondependent variables. The package has been thoroughly tested, and is available in several common computer languages (Pascal, Modula-2 and C). It is easy to use, and advanced knowledge of mathematics or computers is not essential. Standard test problems, and a fully working example of use on enzyme kinetics are included.     

Application of Romero García method to the degradation process of DGEBA/ETDA filled with ultra fine metallic copper

Summary The thermal properties of an epoxy resin (diglycidyl ether Bisphenol A, DGEBA) cured with a primary amine (ethylenediamine, ETDA) and filled with 10, 20 and 30 wt. % of ultra fine copper particles were analyzed. The thermal results were evaluated by means of the Romero-Garcia method, which allowed to obtain the resin degradation kinetic parameters, as well as the possible decomposition mechanism. From the obtained results is possible to infer that higger copper content strongly affects the degradation process of the epoxy resin, which causes a drop off on the composites thermal stability. This behavior could be attributed to water presence, being this effect less pronounced for the resin with 10 wt. % copper and unfilled resin. Regarding to decomposition mechanism, three way transport showed the best correlation for all samples.