High Content Screening of a Kinase-Focused Library Reveals Compounds Broadly-Active against Dengue Viruses

Dengue virus is a mosquito-borne flavivirus that has a large impact in global health. It is considered as one of the medically important arboviruses, and developing a preventive or therapeutic solution remains a top priority in the medical and scientific community. Drug discovery programs for potential dengue antivirals have increased dramatically over the last decade, largely in part to the introduction of high-throughput assays. In this study, we have developed an image-based dengue high-throughput/high-content assay (HT/HCA) using an innovative computer vision approach to screen a kinase-focused library for anti-dengue compounds. Using this dengue HT/HCA, we identified a group of compounds with a 4-(1-aminoethyl)-N-methylthiazol-2-amine as a common core structure that inhibits dengue viral infection in a human liver-derived cell line (Huh-7.5 cells). Compounds CND1201, CND1203 and CND1243 exhibited strong antiviral activities against all four dengue serotypes. Plaque reduction and time-of-addition assays suggests that these compounds interfere with the late stage of viral infection cycle. These findings demonstrate that our image-based dengue HT/HCA is a reliable tool that can be used to screen various chemical libraries for potential dengue antiviral candidates. © 2013 Cruz et al.

Chaotic diffusion caused by close encounters with several massive asteroids: II. the regions of (10) Hygiea, (2) Pallas, and (31) Euphrosyne

Context. Close encounters with (1) Ceres and (4) Vesta, the two most massive bodies in the main belt, are known to be a mechanism of dynamical mobility able to significantly alter proper elements of minor bodies, and they are the main source of dynamical mobility for medium-sized and large asteroids (D > 20 km, approximately). Recently, it has been shown that drift rates caused by close encounters with massive asteroids may change significantly on timescales of 30 Myr when different models (i.e., different numbers of massive asteroids) are considered. Aims. So far, not much attention has been given to the case of diffusion caused by the other most massive bodies in the main belt: (2) Pallas, (10) Hygiea, and (31) Euphrosyne, the third, fourth, and one of the most massive highly inclined asteroids in the main belt, respectively. Since (2) Pallas is a highly inclined object, relative velocities at encounter with other asteroids tend to be high and changes in proper elements are therefore relatively small. It was thus believed that the scattering effect caused by highly inclined objects in general should be small. Can diffusion by close encounters with these asteroids be a significant mechanism of long-term dynamical mobility? Methods. By performing simulations with symplectic integrators, we studied the problem of scattering caused by close encounters with (2) Pallas, (10) Hygiea, and (31) Euphrosyne when only the massive asteroids (and the eight planets) are considered, and the other massive main belt asteroids and non-gravitational forces are also accounted for. Results. By finding relatively small values of drift rates for (2) Pallas, we confirm that orbital scattering by this highly inclined object is indeed a minor effect. Unexpectedly, however, we obtained values of drift rates for changes in proper semi-major axis a caused by (10) Hygiea and (31) Euphrosyne larger than what was previously found for scattering by (4) Vesta. These high rates may have repercussions on the orbital evolution and age estimate of their respective families. © 2013 ESO.

Resting heart rate: Its correlations and potential for screening metabolic dysfunctions in adolescents

Background: In pediatric populations, the use of resting heart rate as a health index remains unclear, mainly in epidemiological settings. The aims of this study were to analyze the impact of resting heart rate on screening dyslipidemia and high blood glucose and also to identify its significance in pediatric populations.Methods: The sample was composed of 971 randomly selected adolescents aged 11 to 17 years (410 boys and 561 girls). Resting heart rate was measured with oscillometric devices using two types of cuffs according to the arm circumference. Biochemical parameters triglycerides, total cholesterol, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol and glucose were measured. Body fatness, sleep, smoking, alcohol consumption and cardiorespiratory fitness were analyzed.Results: Resting heart rate was positively related to higher sleep quality (β = 0.005, p = 0.039) and negatively related to cardiorespiratory fitness (β = -0.207, p = 0.001). The receiver operating characteristic curve indicated significant potential for resting heart rate in the screening of adolescents at increased values of fasting glucose (area under curve = 0.611 ± 0.039 [0.534 - 0.688]) and triglycerides (area under curve = 0.618 ± 0.044 [0.531 - 0.705]).Conclusion: High resting heart rate constitutes a significant and independent risk related to dyslipidemia and high blood glucose in pediatric populations. Sleep and cardiorespiratory fitness are two important determinants of the resting heart rate. © 2013 Fernandes et al.; licensee BioMed Central Ltd.

Note on linearized new massive gravity in arbitrary dimensions

By means of a triple master action we deduce here a linearized version of the new massive gravity (NMG) in arbitrary dimensions. The theory contains a 4th-order and a 2nd-order term in derivatives. The 4th-order term is invariant under a generalized Weyl symmetry. The action is formulated in terms of a traceless ημνΩμνρ=0 mixed symmetry tensor Ωμνρ=-Ωμρν and corresponds to the massive Fierz-Pauli action with the replacement e μν=∂ρΩμνρ. The linearized 3D and 4D NMG theories are recovered via the invertible maps Ωμνρ=Ïμνρβhβμ and Ωμνρ=ÏμνργδT [γδ]μ respectively. The properties h μν=hνμ and T[[γδ]μ]= 0 follow from the traceless restriction. The equations of motion of the linearized NMG theory can be written as zero curvature conditions ∂νTρμ-∂ρT νμ=0 in arbitrary dimensions. © 2013 American Physical Society.

Chiral symmetry, massive gluons and confinement

It is quite difficult to obtain non-trivial chiral symmetry breaking solutions for the quark gap equation in the presence of dynamically generated gluon masses. An effective confining propagator has recently been proposed by Cornwall in order to solve this problem. We study phenomenological consequences of this approach, showing its compatibility with the experimental data. We argue that this confining propagator should be restricted to a small region of momenta, leading to effective four-fermion interactions at low energy. © 2013 American Institute of Physics.

Correspondence between the self-dual model and the topologically massive electrodynamics: A new view

Following the study of the Topologically Massive Theories under the Hamilton-Jacobi, we now analyze the constraint structure of the Self-Dual model as well as its correspondence with the Topologically Massive Electrodynamics. © 2013 American Institute of Physics.

Development and validation of a D-loop mtDNA SNP assay for the screening of specimens in forensic casework

Mitochondrial DNA (mtDNA) analysis is usually a last resort in routine forensic DNA casework. However, it has become a powerful tool for the analysis of highly degraded samples or samples containing too little or no nuclear DNA, such as old bones and hair shafts. The gold standard methodology still constitutes the direct sequencing of polymerase chain reaction (PCR) products or cloned amplicons from the HVS-1 and HVS-2 (hypervariable segment) control region segments. Identifications using mtDNA are time consuming, expensive and can be very complex, depending on the amount and nature of the material being tested. The main goal of this work is to develop a less labour-intensive and less expensive screening method for mtDNA analysis, in order to aid in the exclusion of non-matching samples and as a presumptive test prior to final confirmatory DNA sequencing. We have selected 14 highly discriminatory single nucleotide polymorphisms (SNPs) based on simulations performed by Salas and Amigo (2010) [1] to be typed using SNaPShotTM (Applied Biosystems, Foster City, CA, USA). The assay was validated by typing more than 100 HVS-1/HVS-2 sequenced samples. No differences were observed between the SNP typing and DNA sequencing when results were compared, with the exception of allelic dropouts observed in a few haplotypes. Haplotype diversity simulations were performed using 172 mtDNA sequences representative of the Brazilian population and a score of 0.9794 was obtained when the 14 SNPs were used, showing that the theoretical prediction approach for the selection of highly discriminatory SNPs suggested by Salas and Amigo (2010) [1] was confirmed in the population studied. As the main goal of the work is to develop a screening assay to skip the sequencing of all samples in a particular case, a pair-wise comparison of the sequences was done using the selected SNPs. When both HVS-1/HVS-2 SNPs were used for simulations, at least two differences were observed in 93.2% of the comparisons performed. The assay was validated with casework samples. Results show that the method is straightforward and can be used for exclusionary purposes, saving time and laboratory resources. The assay confirms the theoretic prediction suggested by Salas and Amigo (2010) [1]. All forensic advantages, such as high sensitivity and power of discrimination, as also the disadvantages, such as the occurrence of allele dropouts, are discussed throughout the article. © 2013 Elsevier B.V.

Screening of genomic libraries

Microsatellites, or simple sequence repeats (SSRs), have proven to be an important molecular marker in plant genetics and breeding research. The main strategies to obtain these markers can be through genomic DNA and from expressed sequence tags (ESTs) from mRNA/cDNA libraries. Genetic studies using microsatellite markers have increased rapidly because they can be highly polymorphic, codominant markers and they show heterozygous conserved sequences. Here, we describe a methodology to obtain microsatellite using the enrichment library of DNA genomic sequences. This method is highly efficient to development microsatellite markers especially in plants that do not have available ESTs or genome databases. This methodology has been used to enrich SSR marker libraries in Citrus spp., an important tool to genotype germplasm, to select zygotic hybrids, and to saturate genetic maps in breeding programs. © Springer Science+Business Media, LLC 2013.

Massive spin-2 particle from a rank-2 tensor

Here we obtain all possible second-order theories for a rank-2 tensor which describe a massive spin-2 particle. We start with a general second-order Lagrangian with ten real parameters. The absence of lower-spin modes and the existence of two local field redefinitions leads us to only one free parameter. The solutions are split into three one-parameter classes according to the local symmetries of the massless limit. In the class which contains the usual massive Fierz-Pauli theory, the subset of spin-1 massless symmetries is maximal. In another class where the subset of spin-0 symmetries is maximal, the massless theory is invariant under Weyl transformations and the mass term does not need to fit into the form of the Fierz-Pauli mass term. In the remaining third class neither the spin-1 nor the spin-0 symmetry is maximal and we have a new family of spin-2 massive theories. © 2013 American Physical Society.

Note on the nonuniqueness of the massive Fierz-Pauli theory and spectator fields

It is possible to show that there are three independent families of models describing a massive spin-2 particle via a rank-2 tensor. One of them contains the massive Fierz-Pauli model, the only case described by a symmetric tensor. The three families have different local symmetries in the massless limit and can not be interconnected by any local field redefinition. We show here, however, that they can be related with the help of a decoupled and nondynamic (spectator) field. The spectator field may be either an antisymmetric tensor B μν=-Bνμ, a vector Aμ or a scalar field φ, corresponding to each of the three families. The addition of the extra field allows us to formulate master actions which interpolate between the symmetric Fierz-Pauli theory and the other models. We argue that massive gravity models based on the Fierz-Pauli theory are not expected to be equivalent to possible local self-interacting theories built up on top of the two new families of massive spin-2 models. The approach used here may be useful to investigate dual (nonsymmetric) formulations of higher-spin particles. © 2013 American Physical Society.

Cost-effective composite electrode for the fast voltammetric screening and determination of riboflavin (B2) and pyridoxine (B6) in pharmaceuticals

Different solid composites made by mechanical dispersions of graphite particles into heated paraffin (from 65 to 80% graphite, in mass) were prepared and assessed in order to optimize their use in electrochemical and electroanalytical procedures for bioanalysis. Besides these, composites were also evaluated by thermoanalytical techniques aiming to study their conservation and long-term stability (over eight months without special care), among others. Best results were found at 80% m/m graphite in paraffin. Such electrode combines low-cost, stability, sensitivity, ease of maintenance and clearance, besides the possibilities of manufacture in many different forms and shapes (with or without modifications) and applicability in a wide range of pH. Electrochemical studies by different voltammetric techniques involving vitamins from complex B (riboflavin and pyridoxine) leaded to a better understanding about their electrooxidative processes onto carbon-composite electrodes, specially regarding reversibility and pH-dependence. Data were also acquired and optimized with analytical purposes, being square-wave voltammetry in pH 4.2 chosen by its many advantages. Good linearity between peak responses as function of concentration were reached from 5 to 43 μmol L-1 for riboflavin (peak at -0.257 V) and up to 8.5 × 10-4 mol L -1 for pyridoxine (peak at +1.04 V), best studied conditions; limits of detection (at an S/N of 3) for both analites showed to be circa 1.0 mol L-1. Different commercial samples were analyzed for riboflavin (EMS® complex B syrup) and pyridoxine (Citoneurin 5000 Merck® ampoules) providing 96.6% and 98.7% recoveries, respectively.

In vivo analgesic activity, toxicity and phytochemical screening of the hydroalcoholic extract from the leaves of Psidium cattleianum Sabine

Ethnopharmacological relevance: Psidium cattleianum Sabine is extensively used in Brazilian traditional medicine to treat several diseases including painful disorders. Aim of the study to investigate the toxicity and the possible analgesic activities of the hydroalcoholic extract from the leaves of Psidium cattleianum Sabine (ELPCS), to support its use in folk medicine. To screen the major phytochemical constituents of this extract and evaluate their antioxidant activity. Materials and methods: ELPCS was assessed for its antioxidant activity using the DPPH model. Its analgesic activity was examined using mouse models of acetic acid-induced writhing and hot plate paw licking models. The major phytochemical constituents of the extract were screened; their toxicity on LLC-MK2 mammalian cells was evaluated. Results: ELPCS exhibited significant peripheral analgesic activity at doses of 60, 80, 100, 200 and 400 mg/kg in mice, but it did not display central analgesic activity and not was toxic to LLC-MK2 cell (LD 50>400 μg/mL). The extract exhibited free radical scavenging activity as evidenced by IC 50 values (15.9 μg/mL) obtained by the DPPH method. Phytochemical screening detected flavonoids, saponins, cardiac glycosides, anthraquinones, and tannins. Conclusions: The results of the experimental studies proved the analgesic activity of ELPCS and supported the traditional use of this plant. © 2013 Elsevier B.V. All rights reserved.