Purification, characterisation and inhibition studies of protease from Coriandrum sativum

Protease from coriander leaf (Coriandrum sativum) was evaluated for its ability to detect selected heavy metals using Bradford-protease-casein assay system. Considering the highly polluted environment with heavy metals contributed by industrial wastages and its implications on public health, this present study was dedicated to provide a rapid and sensitive assay for the detection of heavy metals in the environmental samples. The basis of the protein assay using casein as a substrate relies upon the inability of the Bradford reagent to stain polypeptide with less than molecular weight of 2 kDa. Casein that has been stained by the Bradford reagent gives a dark blue color. However, the degradation product is not stained by the reagent and the solution remains brown in color. In the presence of heavy metals that inhibit protease activity, casein would remain undigested and the color would remain blue even after incubation. Optimization studies were carried out for this protease prior to heavy metals inhibition studies. The optimization studies include enzyme concentration, substrate concentration, pH, temperature and time of incubation. The optimum concentration of protease, substrate, temperature and incubation time for protease were 0.45 mg/ml protease, 0.43 mg/ml casein, 35oC and 20 min respectively after a period of heavy metals incubation. This enzyme was then purified through anion exchanger using DEAE- Cellulose column and gel filtration using Agilent ZORBAX column. The molecular weight detected was around 55 kDa. Protease activity obtained from coriander was found to be optimum at pH around 8 to 9.5. For this bioassay, two heavy metals showed inhibition towards enzyme activity at a concentration of 1 mg/l. The inhibition shown by the heavy metals on protease activity were around 40% for mercury and 70% for zinc. The IC50 values of mercury and zinc were 3.22 mg/l and 0.73 mg/l respectively. The limits of detection (LOD) for mercury and zinc were 0.24 mg/l and 0.23 mg/l respectively. The limits of quantitation (LOQ) for mercury and zinc were 0.80 mg/l and 0.76 mg/l respectively. This bioassay using coriander protease was found not to be sensitive towards pesticides and xenobiotics. The advantage of the protease bioassay compared to other bioassay relies on its rapidity, simplicity, economical value, stability in severe conditions such as pH and temperature as well as relatively interference free from detergents, solvents and pesticides

Intelligent Reward based Data Offloading in Next Generation Vehicular Networks

A massive increase in the number of mobile devices and data hungry vehicular network applications creates a great challenge for Mobile Network Operators (MNOs) to handle huge data in cellular infrastructure. However, due to fluctuating wireless channels and high mobility of vehicular users, it is even more challenging for MNOs to deal with vehicular users within a licensed cellular spectrum. Data offloading in vehicular environment plays a significant role in offloading the vehicle s data traffic from congested cellular network s licensed spectrum to the free unlicensed WiFi spectrum with the help of Road Side Units (RSUs). In this paper, an Intelligent Reward based Data Offloading in Next Generation Vehicular Networks (IR-DON) architecture is proposed for dynamic optimization of data traffic and selection of intelligent RSU. Within IR-DON architecture, an Intelligent Access Network Discovery and Selection Function (I-ANDSF) module with Q-Learning, a reinforcement learning algorithm is designed. I-ANDSF is modeled under Software-Defined Network (SDN) controller to solve the dynamic optimization problem by performing an efficient offloading. This increases the overall system throughput by choosing an optimal and intelligent RSU in the network selection process. Simulation results have shown the accurate network traffic classification, optimal network selection, guaranteed QoS, reduced delay and higher throughput achieved by the I-ANDSF module

Near real-time biomonitoring of copper from an industrial complex effluent discharge site using a plant protease inhibitive assay

In this work, a temporal monitoring work for heavy metals from an effluent discharge point in the Juru Industrial Estate was carried out using the protease extracted from garlic (Allium sativum) as the principal bioassay system. Casein-Coomassie-dye binding assay method has utilized this purpose. The periodic sampling results for one day of a location in the Juru Industrial Estate showed temporal variation of copper concentration coinciding with garlic protease inhibition with the highest concentrations of copper occurring between 12.00 and 16.00 hours of between 3 and 3.5 mg/L copper. The crude proteases extracted from Allium sativum successfully detect temporal variation of copper form this location. In conclusion, this assay method has the potential to be a rapid, sensitive, and economic inhibitive assay for the large-scale biomonitoring works for the heavy metal copper from this area

Preliminary screening of plant proteases as a potential source for the development of an inhibitive assay for heavy metals

Heavy metals pollution has become a great threat to the world. Since instrumental methods are expensive and need skilled technician, a simple and fast method is needed to determine the presence of heavy metals in the environment. In this work, a preliminary study was carried out on the applicability of various local plants as a source of protease for the future development of the inhibitive enzyme assay for heavy-metals. The crude proteases preparation was assayed using casein as a substrate in conjunction with the Coomassie dye-binding assay. The crude protease from the kesinai plant was found to be the most potent plant protease. The crude enzyme exhibited broad temperature and pH ranges for activity and will be developed in the future as a potential inhibitive assay for heavy metals

Anti-HMG-CoA reductase, antioxidant, and anti-inflammatory activities of amaranthus viridis leaf extract as a potential treatment for hypercholesterolemia

Inflammation and oxidative stress are believed to contribute to the pathology of several chronic diseases including hypercholesterolemia (elevated levels of cholesterol in blood) and atherosclerosis. HMG-CoA reductase inhibitors of plant origin are needed as synthetic drugs, such as statins, which are known to cause adverse effects on the liver and muscles. Amaranthus viridis (A. viridis) has been used from ancient times for its supposedly medically beneficial properties. In the current study, different parts of A. viridis (leaf, stem, and seed) were evaluated for potential anti-HMG-CoA reductase, antioxidant, and anti-inflammatory activities. The putative HMG-CoA reductase inhibitory activity of A. viridis extracts at different concentrations was determined spectrophotometrically by NADPH oxidation, using HMG-CoA as substrate. A. viridis leaf extract revealed the highest HMG-CoA reductase inhibitory effect at about 71%, with noncompetitive inhibition in Lineweaver-Burk plot analysis. The leaf extract showed good inhibition of hydroperoxides, 2,2-diphenyl-1-picrylhydrazyl (DPPH), nitric oxide (NO), and ferric ion radicals in various concentrations. A. viridis leaf extract was proven to be an effective inhibitor of hyaluronidase, lipoxygenase, and xanthine oxidase enzymes. The experimental data suggest that A. viridis leaf extract is a source of potent antioxidant and anti-inflammatory agent and may modulate cholesterol metabolism by inhibition of HMG-CoA reductase

QoS-Aware Frequency-Based 4G+Relative Authentication Model for Next Generation LTE and Its Dependent Public Safety Networks

Increasing demands for high-speed broadband wireless communications with voice over long term evolution (LTE), video on demand, multimedia, and mission-critical applications for public safety motivate 4th-generation (4G) and 5G communication development. The flat IP-based LTE and LTE-Advanced technologies are the expected key drivers for 5G. However, LTE, with its elapsed security mechanism and open nature, leaves a huge loophole for intruders to jeopardize the entire communication network. The timeand bandwidth-consuming authentication procedure in LTE leads to service disruptions and makes it unfit for public safety applications. To cater the prevailing LTE security and service requirements, we propose the 4G plus relative authentication model (4G+RAM), which is composed of two dependent protocols: 1) Privacy-protected evolved packet system authentication and key agreement protocol for the initial authentication (PEPS-AKA) and 2) 4G plus frequency-based re-authentication protocol for the re-authentication of known and frequent users (4G+FRP). The 4G+RAM supports seamless communication with a minimum signaling load on core elements and conceals users' permanent identifiers to ensure user privacy. We simulate the proposed protocols for formal security verification with the widely accepted automated validation of Internet security protocols and applications tool. A comparative analysis of bandwidth consumption is also performed and proved that the proposed 4G+RAM outperforms the existing solutions

Biodegradation of hydrocarbon sludge by pseudomonas sp.strain UPM-KV

Ahydrocarbon-utilizing microorganism isolated locallywas characterized and investigated. This study involved standard biochemical tests and investigation of the bacterial growth based on the uptake of carbon and nitrogen source, temperature and optimum pH growth. The bacterium was found to be a Gram-negative rod, non-motile property with unique property to degrade hydrocarbon sludge. By using diesel as sole carbon source the bacterium was found to be an aerobe as further proven by the oxidase testing. The optimum conditions for the growth was found to be at 30 °C and pH 6.8 with optimum diesel concentration, 1% (v/v). The bacterium ideally used ammonium sulfate as source of nitrogen and was identified as Pseudomonas sp.strain UPM-KV. The ability of this bacterium to efficiently grow on hydrocarbon sludge makes the bacterium an important tool for bioremediation of this toxic sludge that contains high concentration of heavy metals

Observation of the Bile Canaliculi of Puntius javanicus Liver affected by Copper

Investigation on in vivo effects of copper (Cu) on the ultrastructure of P. javanicus liver was carried out using transmission electron microscopy (TEM). The addition of sublethal concentration of 5 mg/L of Cu caused abnormalities on the bile canaliculi (BC) including dilation and elongation compared to control and at lower concentrations of copper with a normal round shape form. Findings from this study support an alternative histological assessment of the effects of Cu concentration on P. javanicus liver

Identification of marine bacteria isolated from marine soil sediments and their ability to biosynthesise AgNPs extracellularly

Over the past few years, nanoparticles synthesis is one of the most active research in the nanotechnology field. The synthesis can be done chemically, physically and biologically. However, some researchers prefer to synthesise it biologically or also known as biosynthesis or ‘green synthesis’ because it is believed to be safer, environmentally friendly and cost-effective. In this work, we report the extracellular synthesis of 20 isolated marine bacteria from marine soil sediment which were identified and evaluated to synthesise silver nanoparticles (AgNPs). This was done by the addition of silver nitrate (AgNO3) solution with the cell-free supernatant of the isolated marine bacteria at room temperature. The marine bacteria were identified using 16S rRNA identification and neighbour-joining phylogenetic tree were constructed. Identification results showed that the isolated bacteria consist of 19 Serratia sp. and 1 Providencia sp. The biosynthesised AgNPs colloids were evaluated using morphological and optical analysis. AgNPs were observed for colour change and determined using ultraviolet-visible (UV-Vis) spectrophotometer. The existence of surface plasmon resonance peak at 400 to 450 nm is evidence of AgNPs formation