A Conditionally Fluorescent Peptide Reporter of Secondary Structure Modulation

Proteins containing intrinsic disorder often form secondary structure upon interaction with a binding partner. Modulating such structures presents an approach for manipulating the resultant functional outcomes. Translational repressor protein 4E-BP1 is an example of an intrinsically disordered protein that forms an α-helix upon binding to its protein ligand, eIF4E. Current biophysical methods for analyzing binding-induced structural changes are low-throughput, require large amounts of sample, or are extremely sensitive to signal interference by the ligand itself. Herein, we describe the discovery and development of a conditionally fluorescent 4E-BP1 peptide that reports structural changes of its helix in high-throughput format. This reporter peptide is based on conditional quenching of fluorescein by thioamides. In this case, fluorescence signal increases as the peptide becomes more ordered. Conversely, destabilization of the α-helix results in decreased fluorescence signal. The low concentration and low volume of peptide required make this approach amenable for high-throughput screening to discover ligands that alter peptide secondary structure.     

Leaf-to-Whole Plant Spread Bioassay for Pepper and Ralstonia solanacearum Interaction Determines Inheritance of Resistance to Bacterial Wilt for Further Breeding

Bacterial wilt (BW) disease from Ralstonia solanacearum is a serious disease and causes severe yield losses in chili peppers worldwide. Resistant cultivar breeding is the most effective in controlling BW. Thus, a simple and reliable evaluation method is required to assess disease severity and to investigate the inheritance of resistance for further breeding programs. Here, we developed a reliable leaf-to-whole plant spread bioassay for evaluating BW disease and then, using this, determined the inheritance of resistance to R. solanacearum in peppers. Capsicum annuum ‘MC4′ displayed a completely resistant response with fewer disease symptoms, a low level of bacterial cell growth, and significant up-regulations of defense genes in infected leaves compared to those in susceptible ‘Subicho’. We also observed the spreading of wilt symptoms from the leaves to the whole susceptible plant, which denotes the normal BW wilt symptoms, similar to the drenching method. Through this, we optimized the evaluation method of the resistance to BW. Additionally, we performed genetic analysis for resistance inheritance. The parents, F₁ and 90 F₂ progenies, were evaluated, and the two major complementary genes involved in the BW resistance trait were confirmed. These could provide an accurate evaluation to improve resistant pepper breeding efficiency against BW.     

Assessing political second screening behavior and personality traits: The roles of economic development,freedom of expression and monochromatic vs. polychromatic cultures

This study focuses on an emerging media multitasking phenomenon called second screening or dual screening. Employing two-wave panel-data from 19 countries, we test whether the Big Five personality traits help explain the use of an additional screen or device while watching political content on TV to discuss the program with others or to look up for additional information. Results show that extraversion positively predicts political second screening. In contrast, agreeableness and openness to new experience are negatively related to political second screening. Moreover, multilevel analysis is performed to test whether the between-country variation is related to economic, political and cultural indicators.     

A novel GPU-accelerated strategy for contingency screening of static security analysis

Graphics processing unit (GPU) has been applied successfully in many computation and memory intensive realms due to its superior performances in float-pointing calculation, memory bandwidth and power consumption, and has great potential in power system applications. Contingency screening is a major time consuming part of contingency analysis. In the absence of relevant existing research, this paper is the first of its kind to propose a novel GPU-accelerated algorithm for direct current (DC) contingency screening. Adapting actively unique characteristics of GPU software and hardware, the proposed GPU algorithm is optimized from four aspects: data transmission, parallel task allocation, memory access, and CUDA (Compute Unified Device Architecture) stream. Case studies on a 3012-bus system and 8503-bus system have shown that the GPU-accelerated algorithm, in compared with its counterpart CPU implementation, can achieve about 20 and 50 times speedup respectively. This highly promising performance has demonstrated that carefully designed performance tuning in conjunction with GPU programing architecture is imperative for a GPU-accelerated algorithm. The presented performance tuning strategies can be applicable to other GPU applications in power systems.     

Nucleus and cytoplast contour detector from a cervical smear image

In this paper, a nucleus and cytoplast contour detector (NCC detector) is presented to automatically detect the cytoplast and nucleus contours of a cell in a cervical smear image. The NCC detector uses the adaptable threshold decision (ATD) method to separate the cell from the cervical smear image, and then uses the maximal gray-level-gradient-difference (MGLGD) method, proposed in this paper, to extract the nucleus from the cell. The experimental results show that the NCC detector is superior to two existing methods, the gradient vector flow-active contour model (GVF-ACM) and the edge enhancement nucleus and cytoplast contour (ENNCC) detector, in segmenting the cytoplast and nucleus of a cell.     

Development of new high-throughput screening method to compare and to detect efficient catalysts for adhesive materials

Organotin compounds particularly dibutyltin dilaurate, are commonly used as catalysts in coatings or adhesive materials to crosslink silyl modified polymers. However, environmental concerns should lead before 2020 to ban organotin compounds due to their high toxicity. Thus new catalysts must be developed. According to the different types of catalytic systems, i.e. acid, basic or metal containing catalysts, a large variety of candidates should be tested. Thus a high-throughput screening (HTS) method could be an interesting tool for the detection of new efficient catalysts to substitute organotin compounds. We report a global HTS method, compatible with organic amino or acid catalysts libraries, as well as with metal-containing libraries.     

In silico and in vitro screening to identify structurally diverse non-azole CYP51 inhibitors as potent antifungal agent

The problem of resistance to azole class of antifungals is a serious cause of concern to the medical fraternity and thus there is an urgent need to identify non-azole scaffolds with high affinity for lanosterol 14α-demethylase (CYP51). In view of this we have attempted to identify novel non-azole CYP51 inhibitors through the application of pharmacophore based virtual screening and in vitro evaluation. A rigorously validated pharmacophore model comprising of 2 hydrogen bond acceptor and 2 hydrophobic features has been developed and used to mine NCI database. Out of 265 retrieved hits, NSC 1215 and 1520 have been chosen on the basis of Lipinski’s rule of five, fit and estimated values. Both the hits were docked into the active site of CYP51. In view of high fit value and CDocker score, NSC 1215 and 1520 have been subjected to in vitro microbiological assay. The result reveals that NSC 1215 and 1520 are active against Candida albicans, Candida parapsilosis, Candida tropicalis, and Aspergillus niger. In addition to this the absorption characteristics of both the hits have also been determined using the rat sac technique and permeation in order of NSC 1520 > NSC 1215 has been observed.     

Screening for lipid depositor of Indonesian microalgae isolated from seashore and peat-land

The screening for finding the lipid producer of indigenous Indonesian tropical microalgae that obtained from peat-land at Riau, Sumatra and seashore at Lombok has carried out. Preliminary screening was done by selection on faster growing algae from 44 (forty-four) isolates. 10 (ten) isolates were chosen for further analysis. Out of 10 selected isolates shows that five isolates have indication rapid growth and high content of lipid, furthermore those chose isolates were observed comprehensively, namely LIPI11-2-Al005, LIPI11-2-Al010, LIPI11-2-Al015, LIPI11-2-Al018, and LIPI11-2-Al019. The microalgae are belong to the eukaryotic microalgae and its seem to be closed to Chlorophytes that has cell nuclear with an envelope, light green chloroplast, thick cell wall, round and elongated cell features. Cultivation of the microalgae in laboratory condition (1 L) has shown that lipid content was about 20–35% base in cell dry weight. The highest lipid content was found in the isolate LIPI11-2-Al018 of 30.74% per dry weight cell. LIPI11-2-A1018 therefore was test for scale up cultivation reach the culture volume of 5 L in the cylindrical photobioreactor, and expected to be source of triglycerides and lipids for biodiesel ingredient.     

A screening sampling plan to detect Mycobacterium avium subspecies paratuberculosis-positive dairy herds

Mycobacterium avium ssp. paratuberculosis (MAP) is the etiological agent of paratuberculosis, a chronic contagious bacterial disease primarily affecting dairy cattle. Paratuberculosis represents a dual problem for the milk production chain: in addition to economic losses to affected herds, MAP may have zoonotic potential. Infected herds must be identified in order to implement programs designed to reduce the incidence of disease within and between herds and to prevent MAP from entering the food chain. The objective of this study was to evaluate the sensitivity and specificity of a screening sampling plan (SSP) to detect MAP-positive dairy herds by repetitive analysis of bulk tank milk (BTM) samples by ELISA and in-line milk filter (ILMF) samples by PCR. Samples from BTM and ILMF were collected twice from 569 dairy herds in southern Italy. Additionally, 12,016 individual milk samples were collected: 9,509 from 102 SSP-positive herds (SSP MAP-positive) and 2,507 from 21 randomly selected SSP-negative herds (SSP MAP-negative). There was a total of 126 SSP MAP-positive herds (i.e., 21.3% SSP MAP-positive herds; 95% confidence interval = 18.0–24.9); the within-herd apparent prevalence (AP) ranged between 0.00 and 22.73% (mean 6.07%). A significant difference in within-herd AP was shown between SSP MAP-positive herds and SSP MAP-negative herds. A highly significant association was shown between the median AP herd status (>5%) and positivity to at least one ILMF or BTM sample. The SSP detected a minimum of 56.25% of low AP herds (AP ≤2.0%) up to a maximum of 100% of herds with a within-herd AP ≥8.0%. Overall, the SSP detected 85.57% of herds in which at least one individual milk sample was positive by ELISA. The proposed SSP was an inexpensive and useful tool to detect MAP-positive herds with a higher risk of infection diffusion and milk contamination. Although the SSP cannot be used for MAP-free certification of herds, it could be useful to prioritize appropriate control measures aimed at reducing the prevalence of infection in dairy herds and milk contamination.     

The influence of spiral jet-milling on the physicochemical properties of carbamazepine form III crystals: Quality by design approach

The purpose of this study was to investigate the influence of spiral jet-milling process on the physicochemical characteristics of α polymorphic active pharmaceutical ingredient, using Carbamazepine form III as a model drug, and taking into consideration Quality by Design (QbD) approach to pharmaceutical development. A 2^(4-1) factorial screening design was implemented to identify the spiral jet-milling process variables that significantly affect the particle size distribution of milled samples. Diameter of injector nozzles, diameter of ring nozzles and air pressure were selected for further analysis using a 2^(3-1) factorial experimental design. Particle size distribution of additional samples was determined, while physicochemical properties were examined by differential scanning calorimetry (DSC), hot-stage polarized microscopy (HSPM), attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR), and compared to those of un-milled drug. The gathered results shown that applied experimental design approach is capable to predict material behavior and could help in better understanding of material behavior during jet-milling process. Created design space (DS) provides assurance of product quality, expressed as the powder particle sizes lower than 5 μm, as well as, in initial polymorph form existence after jet-milling through combination and interaction of input variables.