Directed enzyme evolution guided by multidimensional analysis of substrate-activity space

The directed evolution of protein function frequently involves identification of mutants with improved properties from a population of variants obtained by mutagenesis. The selection of clones to parent the subsequent generation is crucial to the continued creation of superior progeny. In the present study, multivariate analysis guided the evolution of human glutathione transferase (GST) T1-1 to 65-fold enhanced alkyltransferase activity. Six alternative substrates monitored the substrate-activity space that characterized a mutant library of enzymes, obtained by recombination of DNA and heterologous expression in Escherichia coli. A subset of mutants was identified by their proximity in the targeted region of six-dimensional factor space. DNA from these mutants was recombined to create a new generation of GST variants from which an improved enzyme was isolated. The multidimensional cluster analysis is applicable to quantitative properties in any population of molecules undergoing evolution and can guide the tailoring of proteins, nucleic acids and other chemical structures to novel and improved functions.     

Use of conifer volatiles to reduce injury caused by carrot psyllid,Trioza apicalis,Förster (Homoptera,Psylloidea)

The feeding and oviposition of the carrot psyllid,Trioza apicalis, were reduced by the application of fresh spruce and pine sawdust along the seedling rows in carrot fields. Turpentine and separate monoterpene hydrocarbons, mixed into old sawdust and/or placed in polyethylene tubes, were also effective. At a dose of 0.5 liter/m, fresh sawdust reduced the damage to 18% of the plants, compared to 100% damage in untreated plants. The sawdust materials were spread on the soil surface at four- or seven-day intervals during the oviposition period. The tubes were placed along the carrot rows before the oviposition started. Turpentine and separate monoterpene hydrocarbons afforded a protective effect of the same order of magnitude as that obtained from fresh sawdust. The volatile profiles of the spruce and pine sawdust as well as of the turpentine used were determined.     

Structure–Activity Relationships of Benzoic Acid Derivatives as Antifeedants for the Pine Weevil, Hylobius abietis

Aromatic organic compounds found in the feces of the pine weevil, Hylobius abietis (L.) (Coleoptera: Curculionidae), have been shown to deter feeding behavior in this species, which is a serious pest of planted conifer seedlings in Europe. We evaluated 55 benzoic acid derivatives and a few homologs as antifeedants for H. abietis. Structure–activity relationships were identified by bioassaying related compounds obtained by rational syntheses of functional group analogs and structural isomers. We identified five main criteria of efficiency as antifeedants among the benzoic acid derivatives. By predicting optimal structures for H. abietis antifeedants, we attempted to find a commercial antifeedant to protect conifer seedlings against damage by H. abietis in regenerating forests. New, highly effective antifeedants are methyl 2,4-dimethoxybenzoate, isopropyl 2,4-dimethoxybenzoate, methyl 2-hydroxy-3-methoxybenzoate, methyl (3,5-dimethoxyphenyl)acetate, and methyl (2,5-dimethoxyphenyl)acetate. Of these, methyl 2,4-dimethoxybenzoate and isopropyl 2,4-dimethoxybenzoate have the highest antifeedant indices of all substances tested and are the best candidates for practical applications in order to protect planted seedlings in the field.     

Comparison of methods for estimation of the intravoxel incoherent motion (IVIM) diffusion coefficient (<Emphasis Type="Italic">D</Emphasis>) and perfusion fraction (<Emphasis Type="Italic">f</Emphasis>)

Objective

Intravoxel incoherent motion (IVIM) shows great potential in many applications, e.g., tumor tissue characterization. To reduce image-quality demands, various IVIM analysis approaches restricted to the diffusion coefficient (D) and the perfusion fraction (f) are increasingly being employed. In this work, the impact of estimation approach for D and f is studied.

Materials and methods

Four approaches for estimating D and f were studied: segmented IVIM fitting, least-squares fitting of a simplified IVIM model (sIVIM), and Bayesian fitting of the sIVIM model using marginal posterior modes or posterior means. The estimation approaches were evaluated in terms of bias and variability as well as ability for differentiation between tumor and healthy liver tissue using simulated and in vivo data.

Results

All estimation approaches had similar variability and ability for differentiation and negligible bias, except for the Bayesian posterior mean of f, which was substantially biased. Combined use of D and f improved tumor-to-liver tissue differentiation compared with using D or f separately.

Discussion

The similar performance between estimation approaches renders the segmented one preferable due to lower numerical complexity and shorter computational time. Superior tissue differentiation when combining D and f suggests complementary biologically relevant information.

     

Parental criticism and externalizing behavior problems in adolescents: The role of environment and genotype–environment correlation.

Genetic factors are important for the association between parental negativity and child problem behavior, but it is not clear whether this is due to passive or evocative genotype–environment correlation (rGE). In this study, we applied the extended children-of-twins model to directly examine the presence of passive and evocative rGE as well as direct environmental effects in the association between parental criticism and adolescent externalizing problem behavior. The cross-sectional data come from the Twin and Offspring Study in Sweden (N = 909 pairs of adult twins) and from the Twin Study of Child and Adolescent Development (N = 915 pairs of twin children). The results revealed that maternal criticism was primarily due to evocative rGE emanating from their adolescent's externalizing behavior. On the other hand, fathers' critical remarks tended to affect adolescent problem behavior in a direct environmental way. This suggests that previously reported differences in caretaking between mothers and fathers also are reflected in differences in why parenting is associated with externalizing behavior in offspring. (PsycINFO Database Record (c) 2011 APA, all rights reserved)