Genetically Encoded FRET-Sensor Based on Terbium Chelate and Red Fluorescent Protein for Detection of Caspase-3 Activity

This article describes the genetically encoded caspase-3 FRET-sensor based on the terbium-binding peptide, cleavable linker with caspase-3 recognition site, and red fluorescent protein TagRFP. The engineered construction performs two induction-resonance energy transfer processes: from tryptophan of the terbium-binding peptide to Tb³⁺ and from sensitized Tb³⁺ to acceptor—the chromophore of TagRFP. Long-lived terbium-sensitized emission (microseconds), pulse excitation source, and time-resolved detection were utilized to eliminate directly excited TagRFP fluorescence and background cellular autofluorescence, which lasts a fraction of nanosecond, and thus to improve sensitivity of analyses. Furthermore the technique facilitates selective detection of fluorescence, induced by uncleaved acceptor emission. For the first time it was shown that fluorescence resonance energy transfer between sensitized terbium and TagRFP in the engineered construction can be studied via detection of microsecond TagRFP fluorescence intensities. The lifetime and distance distribution between donor and acceptor were calculated using molecular dynamics simulation. Using this data, quantum yield of terbium ions with binding peptide was estimated.     

Influence of branching regularity on the behavior of hyperbranched polymers in solution

A carbosilane pseudo‐dendrimer with irregular structure and degree of branching DB = 1 was synthesized and fractionated. The molar masses of the obtained fractions ranged from 5000 to 490 000 g mol^?1. The hydrodynamic and conformational properties were studied by the methods of light scattering, sedimentation‐diffusion analysis and viscometry in dilute hexane and toluene solutions. The Mark ? Kuhn ? Houwink dependences were obtained. The results for the pseudo‐dendrimer are compared with the data for the initial hyperbranched poly(diallylcarbosilane) with DB = 0.5 and carbosilane dendrimer. It is shown that the solution properties of the pseudo‐dendrimer are close to those for the irregular hyperbranched polymer and differ significantly from the dendrimer behavior. Hence, it is branching regularity rather than the degree of branching that determines the conformational and hydrodynamic properties of hyperbranched polymers with a high degree of branching DB ≥ 0.5. © 2014 Society of Chemical Industry     

New food safety challenges of viral contamination from a global perspective: Conventional,emerging, and novel methods of viral control

Food- and waterborne viruses, such as human norovirus, hepatitis A virus, hepatitis E virus, rotaviruses, astroviruses, adenoviruses, and enteroviruses, are major contributors to all foodborne illnesses. Their small size, structure, and ability to clump and attach to inanimate surfaces make viruses challenging to reduce or eliminate, especially in the presence of inorganic or organic soils. Besides traditional wet and dry methods of disinfection using chemicals and heat, emerging physical nonthermal decontamination techniques (irradiation, ultraviolet, pulsed light, high hydrostatic pressure, cold atmospheric plasma, and pulsed electric field), novel virucidal surfaces, and bioactive compounds are examined for their potential to inactivate viruses on the surfaces of foods or food contact surfaces (tools, equipment, hands, etc.). Every disinfection technique is discussed based on its efficiency against viruses, specific advantages and disadvantages, and limitations. Structure, genomic organization, and molecular biology of different virus strains are reviewed, as they are key in determining these techniques effectiveness in controlling all or specific foodborne viruses. Selecting suitable viral decontamination techniques requires that their antiviral mechanism of action and ability to reduce virus infectivity must be taken into consideration. Furthermore, details about critical treatments parameters essential to control foodborne viruses in a food production environment are discussed, as they are also determinative in defining best disinfection and hygiene practices preventing viral infection after consuming a food product.     

MicroRNA Profiling of B Cell Subsets from Systemic Lupus Erythematosus Patients Reveals Promising Novel Biomarkers

MicroRNAs control the differentiation and function of B cells, which are considered key elements in the pathogenesis of systemic lupus erythematosus (SLE). However, a common micro(mi)RNA signature has not emerged since published data includes patients of variable ethnic background, type of disease, and organ involvement, as well as heterogeneous cell populations. Here, we aimed at identifying a miRNA signature of purified B cells from renal and non-renal severe SLE patients of Latin American background, a population known to express severe disease. Genome-wide miRNA expression analyses were performed on naive and memory B cells and revealed two categories of miRNA signatures. The first signature represents B cell subset-specific miRNAs deregulated in SLE: 11 and six miRNAs discriminating naive and memory B cells of SLE patients from healthy controls (HC), respectively. Whether the miRNA was up or down-regulated in memory B cells as compared with naive B cells in HC, this difference was abolished in SLE patients, and vice versa. The second signature identifies six miRNAs associated with specific pathologic features affecting renal outcome, providing a further understanding for SLE pathogenesis. Overall, the present work provided promising biomarkers in molecular diagnostics for disease severity as well as potential new targets for therapeutic intervention in SLE.     

Combining Multiple Cues for Visual Madlibs Question Answering

International Journal of Computer Vision - This paper presents an approach for answering fill-in-the-blank multiple choice questions from the Visual Madlibs dataset. Instead of generic and commonly...     

Verification of Hypertorus Communication Grids by Infinite Petri Nets and Process Algebra

A model of a hypertorus communication grid has been constructed in the form of an infinite Petri net. A grid cell represents either a packet switching device or a bioplast cell. A parametric expression is obtained to allow a finite specification of an infinite Petri net. To prove properties of an ideal communication protocol, we derive an infinite Diophantine system of equations from it, which is subsequently solved. Then we present the programs htgen and ht-mcrl2-gen, developed in the C language, which generate Petri net and process algebra models of a hypertorus with a given number of dimensions and grid size. These are the inputs for the respective modeling tools Tina and mCRL2, which provide model visualization, step simulation, state space generation and reduction, and structural analysis techniques. Benchmarks to compare the two approaches are obtained. An ad-hoc induction-like technique on invariants, obtained for a series of generated models, allows the calculation of a solution of the Diophantine system in a parametric form. It is proven that the basic solutions of the infinite system have been found and that the infinite Petri net is bounded and conservative. Some remarks regarding liveness and liveness enforcing techniques are also presented.