Universal mitochondrial 16s rRNA biomarker for mini-barcode to identify fish species in Malaysian fish products

Mislabelling in fish products is a highly significant emerging issue in world fish trade in terms of health and economic concerns. DNA barcoding is an efficient sequencing-based tool for detecting fish species substitution but due to DNA degradation, it is in many cases difficult to amplify PCR products of the full-length barcode marker (~650 bp), especially in severely processed products. In the present study, a pair of universal primers targeting a 198 bp sequence of the mitochondrial 16s rRNA gene was designed for identification of fish species in the processed fish products commonly consumed in Malaysia. The specificity of the universal primers was tested by both in-silico studies using bioinformatics software and through cross-reaction assessment by practical PCR experiments against the DNA from 38 fish species and 22 other non-target species (animals and plants) and found to be specific for all the tested fish species. To eliminate the possibility of any false-negative detection, eukaryotic endogenous control was used during specificity evaluation. The developed primer set was validated with various heat-treated (boiled, autoclaved and microwaved) fish samples and was found to show high stability under all processing conditions. The newly developed marker successfully identified 92% of the tested commercial fish products with 96–100% sequence similarities. This study reveals a considerable degree of species mislabelling (20.8%); 5 out of 24 fish products were found to be mislabelled. The new marker developed in this work is a reliable tool to identify fish species even in highly processed products and might be useful in detecting fish species substitution thus protecting consumers’ health and economic interests.     

Quantitative duplex real-time polymerase chain reaction assay with TaqMan probe detects and quantifies Crocodylus porosus in food chain and traditional medicines

Consumption and exploitation of crocodiles have been rampant for their exotic, nutritive and medicinal attributes. These depredations are alarming and although they have continued to be monitored by wildlife and conservation agencies, unlawful trading of crocodiles shows an increasing trend worldwide. Recently, conventional polymerase chain reaction (PCR) and PCR-restriction fragment length polymorphism (RFLP) assays for crocodile have been documented but they are only suitable for identification and cannot quantify adulterations. We described here a quantitative duplex real-time PCR assay with probes to quantify contributions from Crocodylus porosus materials simultaneously. A very short amplicon size of 127bp was used because longer targets could have been broken down in samples, bringing considerable uncertainty in molecular analysis. We have validated a TaqMan probe-based duplex real-time PCR (qPCR) assay for the detection of 0.004 ng DNA in pure state and 0.1% target meat in model chicken meatball. False negative detection was eliminated through an endogenous control (141-bp site of eukaryotic 18S rRNA). Analysis of 12 model chicken meatballs adulterated with C. porosus reflected 96.3?120.2% target recovery at 0.1?10% adulterations. A validation test of 21 commercial food and traditional medicine (TM) crocodile-based products showed 100% effectiveness. Short amplicon sizes, alternative complementary target, exceptional stability and superior sensitivity suggested the assay could be used for the identification and quantitative determination of C. porosus in any food or TM samples even under degraded conditions.     

Linseed polyurethane/tetraethoxyorthosilane/fumed silica hybrid nanocomposite coatings: Physico-mechanical and potentiodynamic polarization measurements studies

The work presents the physico-mechanical properties and potentiodynamic polarization measurements studies of linseed oil based polyurethane/tetraethoxyorthosilane [LPU/TEOS] hybrid and polyurethane/tetraethoxyorthosilane/fumed silica NC [LPU/TEOS/FS] hybrid nanocomposite coatings. The best coating performance was obtained by the inclusion of 2 wt% FS in 2-LPU/TEOS hybrid. 2-LPU/TEOS/FS produced glossy, transparent, flexibility retentive, scratch-resistant and impact resistant coatings at ambient temperature relative to LPU/TEOS coatings. 2-LPU/TEOS/FS showed good scratch hardness (5.5 kg), impact resistance (250 lb/in.), flexibility (1/8 in.) as investigated by standard methods with corrosion rate obtained as 3.567 × 10⁻⁴ mm/year and 4.05 × 10⁻⁴ mm/year and inhibition efficiency as 99.816% and 99.710% in 3.5% NaOH and 3.5% HCl, respectively.     

Effects of deletion of different PP2C protein phosphatase genes on stress responses in Saccharomyces cerevisiae

A key mechanism of signal transduction in eukaryotes is reversible protein phosphorylation, mediated through protein kinases and protein phosphatases (PPases). Modulation of signal transduction by this means regulates many biological processes. Saccharomyces cerevisiae has 40 PPases, including seven protein phosphatase 2C (PP2C PPase) genes (PTC1–PTC7). However, their precise functions remain poorly understood. To elucidate their cellular functions and to identify those that are redundant, we constructed 127 strains with deletions of all possible combinations of the seven PP2C PPase genes. All 127 disruptants were viable under nutrient‐rich conditions, demonstrating that none of the combinations induced synthetic lethality under these conditions. However, several combinations exhibited novel phenotypes, e.g. the Δptc5Δptc7 double disruptant and the Δptc2Δptc3Δptc5Δptc7 quadruple disruptant exhibited low (13°C) and high (37°C) temperature‐sensitive growth, respectively. Interestingly, the septuple disruptant Δptc1Δptc2Δptc3Δptc4Δptc5Δptc6Δptc7 showed an essentially normal growth phenotype at 37°C. The Δptc2Δptc3Δptc5Δptc7 quadruple disruptant was sensitive to LiCl (0.4 m ). Two double disruptants, Δptc1Δptc2 and Δptc1Δptc4, displayed slow growth and Δptc1Δptc2Δptc4 could not grow on medium containing 1.5 m NaCl. The Δptc1Δptc6 double disruptant showed increased sensitivity to caffeine, congo red and calcofluor white compared to each single deletion. Our observations indicate that S. cerevisiae PP2C PPases have a shared and important role in responses to environmental stresses. These disruptants also provide a means for exploring the molecular mechanisms of redundant PTC gene functions under defined conditions. Copyright © 2014 John Wiley & Sons, Ltd.     

Effect of magnesium content on bioactivity of near invert phosphate-based glasses

A series of phosphate glasses 40P₂O₅-(40−x)CaO-xMgO-(20−y)Na₂O-yTiO₂ (where 0 ≤ x ≤ 24 and y = 0 or 1) with varying MgO contents were investigated for their in vitro calcium phosphate (CaP) formation. Thermal analysis of these glass compositions was conducted and a significant decrease in glass transition temperature from 448°C to 430°C was seen with reducing MgO content from 24 to 8 mol%. Degradation studies were performed in phosphate buffered saline (PBS) at 37°C, where the 8 mol% MgO glass showed the highest mass loss of around 3.4% after 28 days of immersion. Cation release studies were conducted via ion chromatography, using ultrapure water at 37°C as the degradation medium. The highest release of Ca²⁺ and Na⁺ ions was observed with the 8 mol% MgO glass. In vitro CaP formation studies were conducted using glass discs immersed in simulated body fluid (SBF) at 37°C for up to 28 days. The amorphous phase and chemical composition of deposited CaP layers on the glass discs were confirmed via X-ray diffraction (XRD) and energy-dispersive X-ray spectroscopy (EDX) analysis, respectively. CaP layers with Ca/P ratio 0.8-1.1 were found to be deposited on the lower MgO content (8 to 2 mol%) glass surface after 28 days of SBF study.     

Structural,morphological, optical and electrical properties of spray deposited zinc doped copper oxide thin films

Nanostructured spray deposited zinc (Zn) doped copper oxide (CuO) thin films were characterized by employing X-ray diffraction (XRD), field emission scanning electron microscopy (FESEM), energy dispersive X-ray (EDX), atomic force microscopy (AFM) and ultraviolet–visible–near infrared (UV–Vis–NIR) spectroscopy. XRD patterns of CuO and Zn doped CuO thin films indicated monoclinic structure with the preferred orientation along \(\left( {\bar 111} \right)\) plane. Maximum value of crystallite size is found about 28.24 nm for 5 at% Zn doped CuO thin film. In FESEM images, nanoparticles were observed around the nucleation center. EDX analysis confirms the presence of all component elements in CuO and Zn doped CuO thin films. Analysis by AFM of CuO and Zn doped CuO thin films figured out decrease of surface roughness due to Zn doping. UV–Vis–NIR spectroscopy showed that CuO and Zn doped CuO thin films are highly transparent in the NIR region. Optical band gap of CuO thin films decreased with substrate temperature and that of Zn doped CuO thin films increased with Zn concentration. Refractive index of CuO and Zn doped CuO thin films raised with photon wavelength and became constant in the NIR region. 5 at% Zn doped CuO thin film showed the highest optical conductivity and the lowest electrical resistivity at room temperature.     

Structure,thermal properties,dissolution behaviour and biomedical applications of phosphate glasses and fibres: a review

For the last few decades, there has been a growing interest in using glasses for biomedical applications. Bioactive glasses are a group of surface reactive glasses which can initiate a range of biological responses by releasing ions into the local environment. Silicate, borate and phosphate glasses are known to show good bioactive characteristics and could be potentially used as favourable templates for bone-tissue formation. Phosphate glasses are unique group of materials that offer great potential for hard and soft tissue engineering over other types of bioactive glasses due to their fully resorbable characteristics, with some formulations possessing chemical composition similar to the mineral phase of natural bone. Moreover, these phosphate glasses can be prepared as fibres which could be used for soft tissue engineering and as fibrous reinforcement for resorbable polymers such as poly-(lactic acid) for fracture fixation applications. This review details some of the properties of phosphate glasses, such as thermal, viscosity/temperature, dissolution and biocompatibility of and how different factors can effectively alter these properties. The effect of the addition of different modifier oxides on the structure in terms of chain length is included. This review also reports on the manufacturing process, mechanical properties and biomedical application of phosphate glass fibres. A brief comparison between three different types of bioactive glasses has also been presented in this review. The main aim of this review is to present the factors affecting the properties of phosphate glasses and glass fibres and how these may be exploited in the design of a biomaterial.     

Effect of structural nonlinearity on probabilistic risk assessment of offshore wind turbine including inelastic soil medium

This research intends to evaluate the influence of structural nonlinearity on the seismic risk of an offshore wind turbine with respect to linear analysis. The structural nonlinearity is presented into the structure, as materially nonlinear, by calibrating plastic hinge at the end of the elastic beam-element of the structure. To guarantee ideal circumstance for seismic analysis, this study includes inelastic soil stratum by using an equivalent linear approach. Monte Carlo simulation is performed by means of seismic vulnerability of the structural system. The research presents a study of a large amount of simulation over the nonlinear and linear structures considering the random character of basic variables of soil under selective earthquakes. The earthquakes having different source-to-site (STS) distances ranging from 7 to 145?km have consistency with the soil parameters. This rigorous implementation is done to accomplish site-specific dynamic analysis. Illustrative results obtained from nonlinear and linear dynamic analysis are compared. The overall finding shows that the nonlinear structure produces highest estimated uncertainty compared to the linear structure. Another termination can be addressed that the earthquake with less STS distance (7.29?km) causes the highest level of destruction to the structure.     

cFEM: a cluster based feature extraction method for network intrusion detection

International Journal of Information Security - The recent trend in network intrusion detection leverages key features of machine learning (ML) algorithms to detect network traffic anomalies....     

A Low Duty Cycle MAC Protocol for Directional Wireless Sensor Networks

Directional communication in wireless sensor networks minimizes interference and thereby increases reliability and throughput of the network. Hence, directional wireless sensor networks (DWSNs) are fastly attracting the interests of researchers and industry experts around the globe. However, in DWSNs the conventional medium access control protocols face some new challenges including the synchronization among the nodes, directional hidden terminal and deafness problems, etc. For taking the advantages of spatial reusability and increased coverage from directional communications, a low duty cycle directional Medium Access control protocol for mobility based DWSNs, termed as DCD-MAC, is developed in this paper. To reduce energy consumption due to idle listening, duty cycling is extensively used in WSNs. In DCD-MAC, each pair of parent and child sensor nodes performs synchronization with each other before data communication. The nodes in the network schedule their time of data transmissions in such a way that the number of collisions occurred during transmissions from multiple nodes is minimized. The sensor nodes are kept active only when the nodes need to communicate with each other. The DCD-MAC exploits localized information of mobile nodes in a distributed manner and thus it gives weighted fair access of transmission slots to the nodes. As a final point, we have studied the performance of our proposed protocol through extensive simulations in NS-3 and the results show that the DCD-MAC gives better reliability, throughput, end-to-end delay, network lifetime and overhead comparing to the related directional MAC protocols.