Simultaneous fractionation of multiple classes of polyphenols from honeybush tea using solid-phase extraction

The structural diversity of polyphenols and the inherent limitations of current extraction techniques pose a challenge to extract polyphenols using a simple and green method. Hence, in this study, a method was developed to simultaneously fractionate multiple classes of polyphenols by only varying ethanol-water solutions. Honeybush tea, which is rich in polyphenols, was selected as a model for this study. Solvent extraction followed by solid-phase extraction (SPE) was developed to obtain a polyphenol-rich fraction from six honeybush samples. Based on a gradient elution programme (10%, 30%, 50%, 70% and 90% (v/v) ethanol-water solution) of SPE, the Strata X cartridge showed a better recovery of most targeted polyphenols under 0.9 mL of the drying volume and 1 mL min⁻¹ of the dispensing speed. The elution programme for fractionating most polyphenols was as follows: single elution with 50% ethanol, followed by twice elution with 70% ethanol. The antioxidant capacity was used to analyse the differences among the polyphenol-rich fractions from six honeybush samples. Principal component analysis (PCA) revealed that unfermented C. genistoides (GG) has the greatest antioxidant capacity among the honeybush species studied. Additionally, mangiferin, isomangiferin and vicenin-2 were the main contributors to the antioxidant capacity in six honeybush fractions according to the correlation study.     

Numerically optimal design for the system with coupled complex transport phenomena-application to the Submerged Entry<Emphasis Type="Italic">Nozzle</Emphasis> in continuous slab caster

This study presents a simple and robust algorithm for the optimal design of the system with coupled complex transport phenomena: the transport phenomena comprise fluid flow, heat and mass transfer. The (1+1)-Evolution Strategy method is adopted as the optimization method. In order to analyze the transport phenomena in the complex geometry generated during the optimization procedure, thefinite volume method with a boundary fitted curvilinear coordinate system is used. To confirm the validity of the present method, the optimal design for the inner shape of the simplified two-dimensionalSubmerged Entry Nozzle in the continuous slab caster is conducted. It is shown that the resulting design of the nozzle is consistent with the purpose and constraints of the design.     

Bacteria-Responsive Self-Assembly of Antimicrobial Peptide Nanonets for Trap-and-Kill of Antibiotic-Resistant Strains

Bacterial trapping using nanonets is a ubiquitous immune defense mechanism against infectious microbes. These nanonets can entrap microbial cells, effectively arresting their dissemination and rendering them more vulnerable to locally secreted microbicides. Inspired by this evolutionarily conserved anti-infective strategy, a series of 15 to 16 residue-long synthetic β-hairpin peptides is herein constructed with the ability to self-assemble into nanonets in response to the presence of bacteria, enabling spatiotemporal control over microbial killing. Using amyloid-specific K114 assay and confocal microscopy, the membrane components lipoteichoic acid and lipopolysaccharide are shown to play a major role in determining the amyloid-nucleating capacity as triggered by Gram-positive and Gram-negative bacteria respectively. These nanonets displayed both trapping and killing functionalities, hence offering a direct improvement from the trap-only biomimetics in literature. By substituting a single turn residue of the non-amyloidogenic BTT1 peptide, the nanonet-forming BTT1-3A analog is produced with comparable antimicrobial potency. With the same sequence manipulation approach, BTT2-4A analog modified from BTT2 peptide showed improved antimicrobial potency against colistin-resistant clinical isolates. The peptide nanonets also demonstrated robust stability against proteolytic degradation, and promising in vivo efficacy and biosafety profile. Overall, these bacteria-responsive peptide nanonets are promising clinical anti-infective alternatives for circumventing antibiotic resistance.     

Expression and Characterization of Monomeric Recombinant Isocitrate Dehydrogenases from Corynebacterium glutamicum and Azotobacter vinelandii for NADPH Regeneration

The enzymatic transformation of various chemicals, especially using NADPH-dependent hydroxylase, into more soluble and/or high value-added products has steadily garnered increasing attention. However, the industrial application of these NADPH-dependent hydroxylases has been limited due to the high cost of the cofactor NADPH. As an alternative, enzymatic NADPH-regeneration systems have been developed and are frequently used in various fields. Here, we expressed and compared two recombinant isocitrate dehydrogenases (IDHs) from Corynebacterium glutamicum and Azotobacter vinelandii in Escherichia coli. Both enzymes were hyper-expressed in the soluble fraction of E. coli and were single-step purified to apparent homogeneity with yields of more than 850 mg/L. These enzymes also functioned well when paired with NADPH consumption systems. Specifically, NADPH was regenerated from NADP⁺ when an NADPH-consuming cytochrome P450 BM3 from Bacillus megaterium was incorporated. Therefore, both enzymes could be used as alternatives to the commonly used regeneration system for NADPH. These enzymes also have promising potential as genetic fusion partners with NADPH-dependent enzymes due to the monomeric nature of their quaternary structure, thereby resulting in self-sufficient biocatalysts via NADPH regeneration in a single polypeptide with NADPH-dependent activity.     

Evaluation of copy-move forgery detection: datasets and evaluation metrics

Creating copy-move forgery became even easier using a wide range of software and platforms. Many algorithms have been proposed to solve the problem, but each one of those algorithms has its own drawbacks. Researchers face many challenges in developing copy-move detection algorithms, and in this paper, we focus on two challenges. The first is the benchmark dataset, and the second involves evaluation metrics. In this paper, we investigate the available copy-move datasets and their advantages and disadvantages. In addition, we discuss the different metrics that have been used by researchers to evaluate the copy-move forgery detection (CMFD) algorithms. On that basis, we suggest the standard specifications of the appropriate copy-move dataset and the metrics that should be used to evaluate the detection algorithms. The findings of this paper will help researchers evaluate their algorithms effectively and fairly essential for developing reliable algorithms.     

Motion analytics of zebrafish using fine motor kinematics and multi-view trajectory

Zebrafish is a useful animal model for studying human diseases such as muscle disorders. However, manual monitoring of fish motion is time-consuming and prone to subjective variations. In this paper, an automatic fish motion analytics framework is proposed. The proposed framework could be exploited to help validate zebrafish models of transgenic zebrafish that express human genes carrying mutations which lead to muscle disorders, thus affecting their ability to swim normally. To differentiate between wild-type (normal) and transgenic zebrafish, the proposed framework consists of two approaches to exploit discriminative spatial–temporal kinematic features which are extracted to represent zebrafish movements. First, the proposed approach studies precise quantitative measurements of motor movement abnormalities using a camera with the capability to record videos with high frames rates (up to 1,000 frames per second). This differs from previous works, which only tracked each fish as a single point over time. Second, the proposed approach studies multi-view spatial–temporal swimming trajectories. This differs from previous works which typically only considered single-view analysis of fish swimming trajectories. The proposed motion features are then incorporated into a supervised classification approach to identify abnormal fish movements. Experimental results have shown that the proposed approach is capable of differentiating between wild-type and transgenic zebrafish, thus helping to validate the zebrafish models.     

Comparative characterization and cytotoxicity study of TAT-peptide as potential vectors for siRNA and Dicer-substrate siRNA

Recently, a newly discovered Dicer-substrate siRNA (DsiRNA) demonstrates higher potency in gene silencing than siRNA but both suffer from rapid degradation, poor cellular uptake and chemical instability. Therefore, Tat-peptide was exploited to protect and facilitate their delivery into cells. In this study, Tat-peptide was complexed with siRNA or DsiRNA through simple complexation. The physicochemical properties (particle size, surface charge and morphology) of the complexes formed were then characterized. The ability of Tat-peptide to carry and protect siRNA or DsiRNA was determined by UV-Vis spectrophotometry and serum protection assay, respectively. Cytotoxicity effect of these complexes was assessed in V79 cell line. siRNA-Tat complexes had particle size ranged from 186?±?17.8 to 375?±?8.3?nm with surface charge ranged from ?9.3?±?1.0 to +13.5?±?1.0?mV, depending on the Tat-to-siRNA concentration ratio. As for DsiRNA-Tat complexes, the particle size was smaller than the ones complexed with siRNA, ranging from 176?±?8.6 to 458?±?14.7?nm. Their surface charge was in the range of +27.1?±?3.6 to +38.1?±?0.9?mV. Both oligonucleotide (ON) species bound strongly to Tat-peptide, forming stable complexes with loading efficiency of more than 86%. These complexes were relatively non cytotoxic as the cell viability of ～90% was achieved. In conclusion, Tat-peptide has a great potential as siRNA and DsiRNA vector due to the formation of stable complexes with desirable physical characteristics, low toxicity and able to carry high amount of siRNA or DsiRNA.     

Flexible and flame resistant poly(lactic acid)/organomontmorillonite nanocomposites

The PLA/OMMT nanocomposites were produced using a melt compounding technique with isopropylated triaryl phosphate ester flame retardant (FR; 10–30 parts per 100 resin). The flammability of the PLA/OMMT composites was evaluated with an Underwriter Laboratory (UL‐94) vertical burning test, and their char morphology was studied using scanning electron microscopy (SEM). The thermal properties of the PLA/OMMT were characterized with a thermogravimetric analyzer (TGA) and a differential scanning calorimeter (DSC). The thermal analyses showed that adding FR reduced the decomposition onset temperature (T_o) of PLA/OMMT. Both PLA/OMMT/FR20 and PLA/OMMT/FR30 showed excellent flame retardant abilities, earning a V‐0 rating during the UL‐94 vertical burning test. A compact, coherent and continuous protective char layer was formed in the PLA/OMMT/FR nanocomposites. Additionally, the DSC results indicated that the flexibility of the PLA/OMMT composites increased after adding FR due to the FR‐induced plasticization. The impact strength of PLA/OMMT was greatly increased by the addition of FR. Flexible PLA nanocomposites with high flame resistance were successfully produced. © 2014 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2015 , 132, 41253.     

Photoreduction of Shewanella oneidensis Extracellular Cytochromes by Organic Chromophores and Dye‐Sensitized TiO2

The transfer of photoenergized electrons from extracellular photosensitizers across a bacterial cell envelope to drive intracellular chemical transformations represents an attractive way to harness nature's catalytic machinery for solar‐assisted chemical synthesis. In Shewanella oneidensis MR‐1 (MR‐1), trans‐outer‐membrane electron transfer is performed by the extracellular cytochromes MtrC and OmcA acting together with the outer‐membrane‐spanning porin ? cytochrome complex (MtrAB). Here we demonstrate photoreduction of solutions of MtrC, OmcA, and the MtrCAB complex by soluble photosensitizers: namely, eosin Y, fluorescein, proflavine, flavin, and adenine dinucleotide, as well as by riboflavin and flavin mononucleotide, two compounds secreted by MR‐1. We show photoreduction of MtrC and OmcA adsorbed on Ru^II‐dye‐sensitized TiO₂ nanoparticles and that these protein‐coated particles perform photocatalytic reduction of solutions of MtrC, OmcA, and MtrCAB. These findings provide a framework for informed development of strategies for using the outer‐membrane‐associated cytochromes of MR‐1 for solar‐driven microbial synthesis in natural and engineered bacteria.