Polishable and Renewable DNA Hybridization Biosensors

Routine applications of DNA hybridization biosensors are often restricted by the need for regenerating the single-stranded (ss) probe for subsequent reuse. This note reports on a viable alternative to prolonged thermal or chemical regeneration schemes through the mechanical polishing of oligonucleotide-bulk-modified carbon composite electrodes. The surface of these biocomposite hybridization biosensors can be renewed rapidly and reproducibly by a simple extrusion/polishing protocol. The immobilized probe retains its hybridization activity on confinement in the interior of the carbon paste matrix, with the use of fresh surfaces erasing memory effects and restoring the original target response, to allow numerous hybridization/measurement cycles. We expect that such reusable nucleic acid modified composite electrodes can be designed for a wide variety of biosensing applications.

Biophysical Characterization of Interactions Involving Importin-α during Nuclear Import

Proteins containing the classical nuclear localization sequences (NLSs) are imported into the nucleus by the importin-α/β heterodimer. Importin-α contains the NLS binding site, whereas importin-β mediates the translocation through the nuclear pore. We characterized the interactions involving importin-α during nuclear import using a combination of biophysical techniques (biosensor, crystallography, sedimentation equilibrium, electrophoresis, and circular dichroism). Importin-α is shown to exist in a monomeric autoinhibited state (association with NLSs undetectable by biosensor). Association with importin-β (stoichiometry, 1:1; K D = 1.1 × 10 -8 M) increases the affinity for NLSs; the importin-α/β complex binds representative monopartite NLS (simian virus 40 large T-antigen) and bipartite NLS (nucleoplasmin) with affinities (K D = 3.5 × 10 -8 M and 4.8 × 10 -8 M, respectively) comparable with those of a truncated importin-α lacking the autoinhibitory domain (T-antigen NLS, K D = 1.7 × 10 -8 M; nucleoplasmin NLS, K D = 1.4 × 10 -8 M). The autoinhibitory domain (as a separate peptide) binds the truncated importin-α, and the crystal structure of the complex resembles the structure of full-length importin-α. Our results support the model of regulation of nuclear import mediated by the intrasteric autoregulatory sequence of importin-α and provide a quantitative description of the binding and regulatory steps during nuclear import.

A novel biomagnetic instrumentation with four magnetoresistive sensors to evaluate gastric motility.

A novel instrumentation using anisotropic magnetoresistive (AMR) sensors associated with magnetic coils excitation was developed to evaluate gastrointestinal tract motility parameters. The susceptometer has four sensors that were used to measure the gastric activity contractions (GAC) in anaesthetized dogs, its performance was evaluated by manometry with good results.

Immunosensor based on immobilization of antigenic peptide NS5A-1 from HCV and silk fibroin in nanostructured films

The peptide NS5A-1 (PPLLESWKDPDYVPPWHG), derived from hepatitis C virus (HCV) NS5A protein, was immobilized into layer-by-layer (LbL) silk fibroin (SF) films. Deposition was monitored by UV-vis absorption measurements at each bilayer deposited. The interaction SF/peptide film induced secondary structure in NS5A-1 as indicated by fluorescence and circular dichroism (CD) measurements. Voltammetric sensor (SF/NS5A-1) properties were observed when the composite film was tested in the presence of anti-HCV. The peptide-silk fibroin interaction studied here showed new architectures for immunosensors based on antigenic peptides and SF as a suitable immobilization matrix. © 2013 American Chemical Society.

Effect of lactation length adjustment procedures on genetic parameter estimates for buffalo milk yield

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Genetic characterization of fig tree mutants with molecular markers

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Selection strategies for dairy buffaloes: economic and genetic consequences

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Trends of the genetic connectedness measures among Nelore beef cattle herds

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Transmission system expansion planning by an extended genetic algorithm

The paper presents an extended genetic algorithm for solving the optimal transmission network expansion planning problem. Two main improvements have been introduced in the genetic algorithm: (a) initial population obtained by conventional optimisation based methods; (b) mutation approach inspired in the simulated annealing technique, the proposed method is general in the sense that it does not assume any particular property of the problem being solved, such as linearity or convexity. Excellent performance is reported in the test results section of the paper for a difficult large-scale real-life problem: a substantial reduction in investment costs has been obtained with regard to previous solutions obtained via conventional optimisation methods and simulated annealing algorithms; statistical comparison procedures have been employed in benchmarking different versions of the genetic algorithm and simulated annealing methods.

Non-destructive genetic sampling in fish. An improved method for DNA extraction from fish fins and scales

DNA-based studies have been one of the major interests in conservation biology of endangered species and in population genetics. As species and population genetic assessment requires a source of biological material, the sampling strategy can be overcome by non-destructive procedures for DNA isolation. An improved method for obtaining DNA from fish fins and scales with the use of an extraction buffer containing urea and further DNA purification with phenol-chloroform is described. The methodology combines the benefits of a non-destructive DNA sampling and its high efficiency. In addition, comparisons with other methodologies for isolating DNA from fish demonstrated that the present procedure also becomes a very attractive alternative to obtain large amounts of high-quality DNA for use in different molecular analyses. The DNA samples, isolated from different fish species, have been successfully used on random amplified polymorphic DNA (RAPD) experiments, as well as on amplification of specific ribosomal and mitochondrial DNA sequences. The present DNA extraction procedure represents an alternative for population approaches and genetic studies on rare or endangered taxa.

Morphological characterization and reproductive aspects in genetic variability studies of forage peanut

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Genetic characterization of egg weight, egg production and age at first egg in quails

O objetivo neste estudo foi obter estimativas de parâmetros genéticos para as características peso do ovo, produção de ovos em 189 dias de postura e dia do primeiro ovo em codornas de três linhagens de postura e uma de corte. Os dados foram analisados por meio de procedimentos bayesianos usando amostragem de Gibbs. As estimativas de herdabilidade para peso do ovo, produção de ovos em 189 dias de postura e dia do primeiro ovo foram, respectivamente, para a linhagem amarela, 0,31; 0,84 e 0,53; azul, 0,14; 0,82 e 0,60; vermelha, 0,70; 0,96 e 0,75; e de corte, 0,73; 0,96 e 0,72. As correlações genéticas entre peso do ovo e produção de ovos em 189 dias de postura, peso do ovo e dia do primeiro ovo e, produção de ovos em 189 dias de postura e dia do primeiro ovo foram, para amarela, 0,58; -0,77; e -0,90; azul, 0,09; -0,01; e -0,95; vermelha, 0,09; 0,03; e -0,76; e de corte, -0,18; 0,19 e -0,91. A partir das probabilidades de superposição das distribuições posteriories dos parâmetros, as linhagens dividem-se em dois grupos distintos: um com as linhagens amarela e azul e outro com as linhagens vermelha e de corte.

Assessing relationships between genetic evaluations using robust regression with an application to holsteins in Uruguay

Globalization of dairy cattle breeding has created a need for international sire proofs. Some early methods for converting proofs from one population to another are based on simple linear regression. An alternative robust regression method based on the t-distribution is presented, and maximum likelihood and Bayesian techniques for analysis are described, including the situation in which some proofs are missing. Procedures were used to investigate the relationship between Holstein sire proofs obtained by two Uruguayan genetic evaluation programs. The results suggest that conversion equations developed from data including only sires having proofs in both populations can lead to distorted results, relative to estimates obtained using techniques for incomplete data. There was evidence of non-normality of regression residuals, which constitutes an additional source of bias. A robust estimator may not solve all problems, but can provide simple conversion equations that are less sensitive to outlying proofs and to departures from assumptions.

RAPD optimization for genetic studies with Citrulus lanatus and Sesamum indicum genotypes

The Random Amplified Polymorphic DNA (RAPD) technique is powerful for DNA polymorphism determinations and is widely used in research involving different organisms, but it is known that RAPD can be affected by many factors that may result in false positive bands and non-reproducible assays. In this study, we analyzed the effect of several factors such as DNA template, primer and Taq DNA polymerase concentrations to optimize and standardize the RAPD technique for further genetic studies with Citrulus lanattus and Sesamum indicum L. The best combination of DNA, Taq DNA polymerase enzyme and primer concentrations in RAPD amplification procedures for sesame and watermelon genotypes was established.

Balanced anesthesia with sevoflurane does not alter redox status in patients undergoing surgical procedures

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)