A rapid, efficient, and economical inverse polymerase chain reaction-based method for generating a site saturation mutant library

With the development of deep sequencing methodologies, it has become important to construct site saturation mutant (SSM) libraries in which every nucleotide/codon in a gene is individually randomized. We describe methodologies for the rapid, efficient, and economical construction of such libraries using inverse polymerase chain reaction (PCR). We show that if the degenerate codon is in the middle of the mutagenic primer, there is an inherent PCR bias due to the thermodynamic mismatch penalty, which decreases the proportion of unique mutants. Introducing a nucleotide bias in the primer can alleviate the problem. Alternatively, if the degenerate codon is placed at the 5' end, there is no PCR bias, which results in a higher proportion of unique mutants. This also facilitates detection of deletion mutants resulting from errors during primer synthesis. This method can be used to rapidly generate SSM libraries for any gene or nucleotide sequence, which can subsequently be screened and analyzed by deep sequencing. (C) 2013 Elsevier Inc. All rights reserved.

VIRTUAL VIEW SYNTHESIS BY NON-LOCAL MEANS FILTERING USING TEMPORAL DATA

A novel algorithm for Virtual View Synthesis based on Non-Local Means Filtering is presented in this paper. Apart from using the video frames from the nearby cameras and the corresponding per-pixel depth map, this algorithm also makes use of the previously synthesized frame. Simple and efficient, the algorithm can synthesize video at any given virtual viewpoint at a faster rate. In the process, the quality of the synthesized frame is not compromised. Experimental results prove the above mentioned claim. The subjective and objective quality of the synthesized frames are comparable to the existing algorithms.

Two techniques to improve the precision of a demand-driven null-dereference verification approach

The problem addressed in this paper is sound, scalable, demand-driven null-dereference verification for Java programs. Our approach consists conceptually of a base analysis, plus two major extensions for enhanced precision. The base analysis is a dataflow analysis wherein we propagate formulas in the backward direction from a given dereference, and compute a necessary condition at the entry of the program for the dereference to be potentially unsafe. The extensions are motivated by the presence of certain ``difficult'' constructs in real programs, e.g., virtual calls with too many candidate targets, and library method calls, which happen to need excessive analysis time to be analyzed fully. The base analysis is hence configured to skip such a difficult construct when it is encountered by dropping all information that has been tracked so far that could potentially be affected by the construct. Our extensions are essentially more precise ways to account for the effect of these constructs on information that is being tracked, without requiring full analysis of these constructs. The first extension is a novel scheme to transmit formulas along certain kinds of def-use edges, while the second extension is based on using manually constructed backward-direction summary functions of library methods. We have implemented our approach, and applied it on a set of real-life benchmarks. The base analysis is on average able to declare about 84% of dereferences in each benchmark as safe, while the two extensions push this number up to 91%. (C) 2014 Elsevier B.V. All rights reserved.

Regioselective Synthesis of Nitrofuran Containing Novel Spiropyrrolidine Library through 1,3-Dipolar Cycloaddition Reactions

A novel series of nitrofuran containing spiropyrrolidines has been synthesized with high regioselectivity in moderate to excellent yields via 1,3-dipolar cycloaddition reaction of azomethine ylides with various substituted chalcones.

Lead selection and characterization of antitubercular compounds using the Nested Chemical Library

Discovering new drugs to treat tuberculosis more efficiently and to overcome multidrug resistance is a world health priority. To find novel antitubercular agents several approaches have been used in various institutions worldwide, including target-based approaches against several validated mycobacterial enzymes and phenotypic screens. We screened more than 17,000 compounds from Vichem's Nested Chemical Library(TM) using an integrated strategy involving whole cell-based assays with Corynebacterium glutamicum and Mycobacterium tuberculosis, and target-based assays with protein kinases PknA, PknB and PknG as well as other targets such as PimA and bacterial topoisomerases simultaneously. With the help of the target-based approach we have found very potent hits inhibiting the selected target enzymes, but good minimal inhibitory concentrations (MIC) against M. tuberculosis were not achieved. Focussing on the whole cell-based approach several potent hits were found which displayed minimal inhibitory concentrations (MIC) against M. tuberculosis below 10 mu M and were non-mutagenic, non-cytotoxic and the targets of some of the hits were also identified. The most active hits represented various scaffolds. Medicinal chemistry-based lead optimization was performed applying various strategies and, as a consequence, a series of novel potent compounds were synthesized. These efforts resulted in some effective potential antitubercular lead compounds which were confirmed in phenotypic assays. (C) 2015 Elsevier Ltd. All rights reserved.

Combinatorial selection of molecular conformations and supramolecular synthons in quercetin cocrystal landscapes: a route to ternary solids

The crystallization of 28 binary and ternary cocrystals of quercetin with dibasic coformers is analyzed in terms of a combinatorial selection from a solution of preferred molecular conformations and supramolecular synthons. The crystal structures are characterized by distinctive O-H center dot center dot center dot N and O-H center dot center dot center dot O based synthons and are classified as nonporous, porous and helical. Variability in molecular conformation and synthon structure led to an increase in the energetic and structural space around the crystallization event. This space is the crystal structure landscape of the compound and is explored by fine-tuning the experimental conditions of crystallization. In the landscape context, we develop a strategy for the isolation of ternary cocrystals with the use of auxiliary template molecules to reduce the molecular and supramolecular `confusion' that is inherent in a molecule like quercetin. The absence of concomitant polymorphism in this study highlights the selectivity in conformation and synthon choice from the virtual combinatorial library in solution.

Asymptotic behavior and interpretation of virtual states: The effects of confinement and of basis sets

A real-space high order finite difference method is used to analyze the effect of spherical domain size on the Hartree-Fock (and density functional theory) virtual eigenstates. We show the domain size dependence of both positive and negative virtual eigenvalues of the Hartree-Fock equations for small molecules. We demonstrate that positive states behave like a particle in spherical well and show how they approach zero. For the negative eigenstates, we show that large domains are needed to get the correct eigenvalues. We compare our results to those of Gaussian basis sets and draw some conclusions for real-space, basis-sets, and plane-waves calculations. (C) 2016 AIP Publishing LLC.

Hacia la aldea virtual : evolución de las tecnologías comunicativas y de los modelos antropológicos

Estudiar los medios de comunicación significa estudiar al hombre, la sociedad en la que vive, su evolución y las perspectivas de desarrollo futuro. Después de haber puntualizado brevemente los avances tecnológicos salientes en la historia del hombre y de haber identificado las características más relevantes de las diversas redefiniciones antropológicas que siguieron a la introducción de algunos nuevos medios, nos detendremos principalmente en el estudio de las peculiaridades de los instrumentos de comunicación de masa más modernos, de la fotografía al cine, de la televisión a Internet. El objetivo principal de este aporte, lejos de querer proveer sistemas teóricos definitivos, es trazar un conjunto coherente de temáticas capaces de suscitar nuevas y proficuas cuestiones y de promover un debate abierto a los más diversos aportes, con particular atención a los avances de las comunicaciones telemáticas.

Enfoques de enseñanza presencial y de enseñanza mixta virtual-presencial de profesores universitarios de ambas modalidades

Resumen: El objetivo de este estudio fue analizar los enfoques de las concepciones de enseñanza en aula y de enseñanza mixta de profesores de estas modalidades educativas. El diseño fue no experimental, descriptivo, transversal y corte mixto. La muestra fue de 183 profesores, 129 de cursos cara a cara y 54 de cursos mixtos que respondieron a preguntas abiertas relacionadas con el significado de la enseñanza, del rol del profesor, del rol del estudiante y de estrategias de enseñanza de cursos cara a cara y mixtos. Los resultados mostraron que en la concepción de enseñanza cara a cara y sus dimensiones de rol del profesor y rol del estudiante en profesores de cursos cara a cara y mixtos la tendencia mayor fue hacia el enfoque en aprendizaje. La concepción de enseñanza de cursos mixtos en profesores de las dos modalidades educativas se ubicó fuertemente en un enfoque en enseñanza a diferencia de sus dimensiones que se focalizaron en el aprendizaje. A partir de este dato se sugiere continuar con la investigación en el tema y la inclusión de las concepciones de enseñanza en los cursos de formación docente