Search results for: enzymatic polymerization
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 654

Search results for: enzymatic polymerization

294 Correlation between Diabetic Cataract, HBA1C and Gurakhu, a Clinical Study in Chhattisgarh State

Authors: A. Bhattacharya, Sanjay Gupta, S. H. Bodakhe

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HbA1c is form of the haemoglobin that is used to measure the average plasma glucose concentration over prolonged periods of time. It is formed in a non-enzymatic glycation pathway by hemoglobin's exposure to plasma glucose. In diabetes mellitus, higher amounts of glycated hemoglobin, indicating poorer control of blood glucose levels, have been associated with cardiovascular disease, nephropathy, and retinopathy. Guraku’s basic components are nicotine and jaggery, jaggery is made up of sugarcane so can have a diabetogenic potential which is exacerbated in presence of nicotine. This work had done with the aim to find correlation between Diabetic cataract, HbA1c and Guraku. Subjects were enrolled according to the inclusion and exclusion criteria. In this study total 75 subjects were included. In the study it was found that people consuming Guraku had a high level of HbA1c thus are more prone to the development of diabetic cataract. Male subjects are the more than female subjects. Most of the subjects belong to the lower socioeconomical class and not very educated. It could be concluded that this type of study could be useful in indentifying number of subjects suffering from diabetic cataract whose condition get worse by use of nicotine product like Guraku and preventive measure to be taken in prevention of this type of diabetic complication.

Keywords: diabetic cataract, HbA1c, Guraku, diabetogenic potential

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293 Microfabrication and Non-Invasive Imaging of Porous Osteogenic Structures Using Laser-Assisted Technologies

Authors: Irina Alexandra Paun, Mona Mihailescu, Marian Zamfirescu, Catalin Romeo Luculescu, Adriana Maria Acasandrei, Cosmin Catalin Mustaciosu, Roxana Cristina Popescu, Maria Dinescu

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A major concern in bone tissue engineering is to develop complex 3D architectures that mimic the natural cells environment, facilitate the cells growth in a defined manner and allow the flow transport of nutrients and metabolic waste. In particular, porous structures of controlled pore size and positioning are indispensable for growing human-like bone structures. Another concern is to monitor both the structures and the seeded cells with high spatial resolution and without interfering with the cells natural environment. The present approach relies on laser-based technologies employed for fabricating porous biomimetic structures that support the growth of osteoblast-like cells and for their non-invasive 3D imaging. Specifically, the porous structures were built by two photon polymerization –direct writing (2PP_DW) of the commercially available photoresists IL-L780, using the Photonic Professional 3D lithography system. The structures consist of vertical tubes with micrometer-sized heights and diameters, in a honeycomb-like spatial arrangement. These were fabricated by irradiating the IP-L780 photoresist with focused laser pulses with wavelength centered at 780 nm, 120 fs pulse duration and 80 MHz repetition rate. The samples were precisely scanned in 3D by piezo stages. The coarse positioning was done by XY motorized stages. The scanning path was programmed through a writing language (GWL) script developed by Nanoscribe. Following laser irradiation, the unexposed regions of the photoresist were washed out by immersing the samples in the Propylene Glycol Monomethyl Ether Acetate (PGMEA). The porous structures were seeded with osteoblast like MG-63 cells and their osteogenic potential was tested in vitro. The cell-seeded structures were analyzed in 3D using the digital holographic microscopy technique (DHM). DHM is a marker free and high spatial resolution imaging tool, where the hologram acquisition is performed non-invasively i.e. without interfering with the cells natural environment. Following hologram recording, a digital algorithm provided a 3D image of the sample, as well as information about its refractive index, which is correlated with the intracellular content. The axial resolution of the images went down to the nanoscale, while the temporal scales ranged from milliseconds up to hours. The hologram did not involve sample scanning and the whole image was available in one frame recorded going over 200μm field of view. The digital holograms processing provided 3D quantitative information on the porous structures and allowed a quantitative analysis of the cellular response in respect to the porous architectures. The cellular shape and dimensions were found to be influenced by the underlying micro relief. Furthermore, the intracellular content gave evidence on the beneficial role of the porous structures in promoting osteoblast differentiation. In all, the proposed laser-based protocol emerges as a promising tool for the fabrication and non-invasive imaging of porous constructs for bone tissue engineering. Acknowledgments: This work was supported by a grant of the Romanian Authority for Scientific Research and Innovation, CNCS-UEFISCDI, project PN-II-RU-TE-2014-4-2534 (contract 97 from 01/10/2015) and by UEFISCDI PN-II-PT-PCCA no. 6/2012. A part of this work was performed in the CETAL laser facility, supported by the National Program PN 16 47 - LAPLAS IV.

Keywords: biomimetic, holography, laser, osteoblast, two photon polymerization

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292 Insights on Nitric Oxide Interaction with Phytohormones in Rice Root System Response to Metal Stress

Authors: Piacentini Diego, Della Rovere Federica, Fattorini Laura, Lanni Francesca, Cittadini Martina, Altamura Maria Maddalena, Falasca Giuseppina

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Plants have evolved sophisticated mechanisms to cope with environmental cues. Changes in intracellular content and distribution of phytohormones, such as the auxin indole-3-acetic acid (IAA), have been involved in morphogenic adaptation to environmental stresses. In addition to phytohormones, plants can rely on a plethora of small signal molecules able to promptly sense and transduce the stress signals, resulting in morpho/physiological responses thanks also to their capacity to modulate the levels/distribution/reception of most hormones. Among these signaling molecules, nitrogen monoxide (nitric oxide – NO) is a critical component in several plant acclimation strategies to both biotic and abiotic stresses. Depending on its levels, NO increases plant adaptation by enhancing the enzymatic or non-enzymatic antioxidant systems or by acting as a direct scavenger of reactive oxygen/nitrogen (ROS/RNS) species produced during the stress. In addition, exogenous applications of NO-specific donor compounds showed the involvement of the signal molecule in auxin metabolism, transport, and signaling, under both physiological and stress conditions. However, the complex mechanisms underlying NO action in interacting with phytohormones, such as auxins, during metal stress responses are still poorly understood and need to be better investigated. Emphasis must be placed on the response of the root system since it is the first plant organ system to be exposed to metal soil pollution. The monocot Oryza sativa L. (rice) has been chosen given its importance as a stable food for some 4 billion people worldwide. In addition, increasing evidence has shown that rice is often grown in contaminated paddy soils with high levels of heavy metal cadmium (Cd) and metalloid arsenic (As). The facility through which these metals are taken up by rice roots and transported to the aerial organs up to the edible caryopses makes rice one of the most relevant sources of these pollutants for humans. This study aimed to evaluate if NO has a mitigatory activity in the roots of rice seedlings against Cd or As toxicity and to understand if this activity requires interactions with auxin. Our results show that exogenous treatments with the NO-donor SNP alleviate the stress induced by Cd, but not by As, in in-vitro-grown rice seedlings through increased intracellular root NO levels. The damages induced by the pollutants include root growth inhibition, root histological alterations and ROS (H2O2, O2●ˉ), and RNS (ONOOˉ) production. Also, SNP treatments mitigate both the root increase in root IAA levels and the IAA alteration in distribution monitored by the OsDR5::GUS system due to the toxic metal exposure. Notably, the SNP-induced mitigation of the IAA homeostasis altered by the pollutants does not involve changes in the expression of OsYUCCA1 and ASA2 IAA-biosynthetic genes. Taken together, the results highlight a mitigating role of NO in the rice root system, which is pollutant-specific, and involves the interaction of the signal molecule with both IAA and brassinosteroids at different (i.e., transport, levels, distribution) and multiple levels (i.e., transcriptional/post-translational levels). The research is supported by Progetti Ateneo Sapienza University of Rome, grant number: RG120172B773D1FF

Keywords: arsenic, auxin, cadmium, nitric oxide, rice, root system

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291 Assessment of the Performance of Fly Ash Based Geo-Polymer Concrete under Sulphate and Acid Attack

Authors: Talakokula Visalakshi

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Concrete is the most commonly used construction material across the globe, its usage is second only to water. It is prepared using ordinary Portland cement whose production contributes to 5-8% of total carbon emission in the world. On the other hand the fly ash by product from the power plants is produced in huge quantities is termed as waste and disposed in landfills. In order to address the above issues mentioned, it is essential that other forms of binding material must be developed in place of cement to make concrete. The geo polymer concrete is one such alternative developed by Davidovits in 1980’s. Geopolymer do not form calcium-silicate hydrates for matrix formation and strength but undergo polycondensation of silica and alumina precursors to attain structural strength. Its setting mechanism depends upon polymerization rather than hydration. As a result it is able to achieve its strength in 3-5 days whereas concrete requires about a month to do the same. The objective of this research is to assess the performance of geopolymer concrete under sulphate and acid attack. The assessment is done based on the experiments conducted on geopolymer concrete. The expected outcomes include that if geopolymer concrete is more durable than normal concrete, then it could be a competitive replacement option of concrete and can lead to significant reduction of carbon foot print and have a positive impact on the environment. Fly ash based geopolymer concrete offers an opportunity to completely remove the cement content from concrete thereby making the concrete a greener and future construction material.

Keywords: fly ash, geo polymer, geopolymer concrete, construction material

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290 Assessment of Cassava Varieties in Ecuador for the Production of Lactic Acid From Starch by-Products

Authors: Pedro Maldonado-Alvarado

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An important cassava quality production was detected in Ecuador. However, in this country, few products with low adding-value are produced from the tuber and none from cassava by-products. To our best knowledge, lactic acid was produced from Ecuadorian cassava bagasse starch in a biotechnological way. The objective of this contribution was to study the influence of the fermentation variables (pH and agitation) on the lactic acid production of Ecuadorian cassava varieties from bagasse starch. Enzymatic hydrolysis of cassava bagasse starch for INIAP 650 and INIAP 651 varieties spread in Ecuador was performed using α-amylase and amyloglucosidase. Then, glucose was fermented by Lactobacillus leichmannii strains in different conditions of agitation (0 and 150 rpm) and pH (4.5, 5.0, and 5.5). Significant differences in ash, fibre, protein, lipids, and amylose were found in cassava bagasse starch of INIAP 650 and INIAP 651 with 1.4 and 1.3%, 4.3 and 6%, 1.2 and 2.1%, 1.9 and 1.5%, and 24.3 and 26.5%, respectively. The determination of lactic acid was performed by potentiometric and FTIR analysis. Conversions of cassava bagasse to reduced sugars were 71.7 and 85.1% for INIAP 650 and INIAP 651, respectively. The best lactic acid concentrations were 27.6 and 33.5 g/L, obtained at agitation 150 rpm and pH 5.5 for INIAP 650 and INIAP 651. Qualitative analysis conducted by FTIR spectrophotometry confirmed the presence of lactic acid in the reacted products. This investigation could contribute to the valorisation of residues from promising cassava varieties in Ecuador and hence to increase the development of this country.

Keywords: bagasse starch, cassava, Ecuador, fermentation, lactic acid

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289 Nanoindentation and Physical Properties of Polyvinyl Chloride/Styrene Co-Maleic Anhydride Blend Reinforced by Organo-Bentonite

Authors: D. E. Abulyazied, S. M. Mokhtar, A. M. Motawie

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Polymer blends represent an important class of materials in engineering applications. The incorporation of clay nanofiller may provide new opportunities for this type of materials to enhance their applications. This article reports on the effects of clay on the structure and properties of polymer blends nanocomposites, based on Polyvinyl chloride PVC and styrene co-maleic anhydride SMA blend. Modification of the Egyptian Bentonite EB was carried out using organo-modifier namely; octadecylamine ODA. Before the modification, the cation exchange capacity CEC of the EB was measured. The octadecylamine bentonite ODA-B was characterized using Fourier transform infrared Spectroscopy FTIR, X-Ray Diffraction XRD, and Transition Electron Microscope TEM. A blend of Polyvinyl chloride PVC and styrene co-maleic anhydride SMA (50:50) was prepared in Tetra Hydro Furan (THF). Then nanocomposites of PVC/SMA/ODA-B were prepared by solution intercalation polymerization from 0.50% up to 5% by weight of ODA-B. The nanocomposites are characterized by XRD, TEM. Thermal, nanoindentation, swelling and electrical properties of the nanocomposites were measured. The morphology of the nanocomposites showed that ODA-B achieved good dispersion in the PVC/SMA matrix. Incorporation of 0.5 %, 1%, 3% and 5% by weight nanoclay into the PVC/SMA blends results in an improvement in nanohardness of 16%, 76%, 92%, and 68% respectively. The elastic modulus increased from 4.59 GPa for unreinforced PVC/SMA blend to 6.30 GPa (37% increase) with the introduction of 3% by weight nanoclay. The cross-link density of the nanocomposites increases with increasing the content of ODA-B.

Keywords: PVC, SMA, nanocomposites, nanoindentation, organo-bentonite

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288 Rechargable N-Halamine Nanoparticles for Antibacterial and Antifouling Applications

Authors: Michal Natan, Ori Gutman, Shlomo Margel, Ehud Banin

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Biofilm formation is a serious problem in medical and industrial settings due to the increased resistance of these communities to killing compared to free-living bacteria. This has prompted the search for agents that can inhibit both bacterial growth and biofilm formation. In this study, N-halamine rechargeable nanoparticles (NPs) were synthesized by co-polymerization of the monomer methacryl amide and the cross-linker monomer N,N-methylene bisacryl amide, and were subsequently loaded with Cl+, using bleach. The chlorinated NPs exhibited remarkable stability to organic reagents. The antibacterial mechanism of the P(MAA-MBAA)-Cl NPs involved generation of reactive oxygen species (ROS) only upon exposure to organic media, but not upon incubation in water, suggesting a specific activation. Moreover, a unique interaction of the P(MAA-MBAA)-Cl NPs with Staphylococcus aureus bacteria but not with human cells was discovered, whereby these microorganisms were all specifically targeted and marked for destruction. Finally, in collaboration with Netafim Ltd. irrigation drippers containing the P(MAA-MBAA)-Cl were incubated in the field and were shown to prevent fouling on them for 5 months as opposed to the control drippers that exhibited substantial fouling. Further, the NPs offer recharging to the surface, thus providing long-lasting protection that does not exist in the products available today. Taken together, the results demonstrate the great potential of implementing the charged NPs in devices and surfaces to prevent bacterial growth.

Keywords: bacteria, biofilm, fouling, nanoparticles

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287 The Cellular Internalization Mechanisms of Cationic Niosomes/DNA Complex in HeLa Cells

Authors: Orapan Paecharoenchai, Tanasait Ngawhirunpat, Theerasak Rojanarata, Auayporn Apirakaramwong, Praneet Opanasopit

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Cationic niosomes formulated with Span20, cholesterol and novel synthesized spermine-cationic lipids (2-hydrocarbon tail and 4- hydrocarbon tail) in a molar ratio of 2.5:2.5:1 can mediate high gene transfection in vitro. However, the uptake mechanisms of these systems are not well clarified. In the present study, effect of endocytic inhibitors on the transfection efficiency of niosomes/DNA complexes was determined on a human cervical carcinoma cell line (HeLa cells) using the inhibitors of macropinocytosis (wortmannin), clathrin- and caveolae-mediated endocytosis (methyl-β-cyclodextrin), clathrin-mediated endocytosis (chlorpromazine), caveolae-mediated endocytosis (genistein and filipin), cytosolic transfer (ammonium chloride) and microtubules polymerization (nocodazole). The transfection of niosomes with 2-hydrocarbon tail lipid was blocked by nocodazole, genistein, ammonium chloride and filipin, respectively, whereas, the transfection of niosomes with 4-hydrocarbon tail lipid was blocked by nocodazole, genistein, ammonium chloride, methyl-β-cyclodextrin and filipin, respectively. It can be concluded that these niosomes/DNA complexes were internalized predominantly by endocytosis via clathrin and caveolae-independent pathway.

Keywords: cellular internalization, cationic niosomes, gene carriers, spermine-cationic lipids

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286 Production and Evaluation of Mango Pulp by Using Ohmic Heating Process

Authors: Sobhy M. Mohsen, Mohamed M. El-Nikeety, Tarek G. Mohamed, Michael Murkovic

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The present work aimed to study the use of ohmic heating in the processing of mango pulp comparing to conventional method. Mango pulp was processed by using ohmic heating under the studied suitable conditions. Physical, chemical and microbiological properties of mango pulp were studied. The results showed that processing of mango pulp by using either ohmic heating or conventional method caused a decrease in the contents of TSS, total carbohydrates, total acidity, total sugars (reducing and non-reducing sugar) and an increase in phenol content, ascorbic acid and carotenoids compared to the conventional process. The increase in electric conductivity of mango pulp during ohmic heating was due to the addition of some electrolytes (salts) to increase the ions and enhance the process. The results also indicate that mango pulp processed by ohmic heating contained more phenols, carbohydrates and vitamin C and less HMF compared to that produced by conventional one. Total pectin and its fractions had slightly reduced by ohmic heating compared to conventional method. Enzymatic activities showed a reduction in poly phenoloxidase (PPO) and polygalacturonase (PG) activity in mango pulp processed by conventional method. However, ohmic heating completely inhibited PPO and PG activities.

Keywords: ohmic heating, mango pulp, phenolic, sarotenoids

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285 Future Applications of 4D Printing in Dentistry

Authors: Hosamuddin Hamza

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The major concept of 4D printing is self-folding under thermal and humidity changes. This concept relies on understanding how the microstructures of 3D-printed models can undergo spontaneous shape transformation under thermal and moisture changes. The transformation mechanism could be achieved by mixing, in a controllable pattern, a number of materials within the printed model, each with known strain/shrinkage properties. 4D printing has a strong potential to be applied in dentistry as the technology could produce dynamic and adaptable materials to be used as functional objects in the oral environment under the continuously changing thermal and humidity conditions. The motion criteria could override the undesired dimensional changes, thermal instability, polymerization shrinkage and microleakage. 4D printing could produce restorative materials being self-adjusted spontaneously without further intervention from the dentist or patient; that is, the materials could be capable of fixing its failed portions, compensating for some lost tooth structure, while avoiding microleakage or overhangs at the margins. In prosthetic dentistry, 4D printing could provide an option to manage the influence of bone and soft tissue imbalance during mastication (and at rest) with high predictability of the type/direction of forces. It can also produce materials with better fitting and retention characteristics than conventional or 3D-printed materials. Nevertheless, it is important to highlight that 4D-printed objects, having dynamic properties, could provide some cushion as they undergo self-folding compensating for any thermal changes or mechanical forces such as traumatic forces.

Keywords: functional material, self-folding material, 3D printing, 4D printing

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284 Evaluation of Hollocelulase Production for Lignocellulosic Biomass Degradation by Penicillium polonicum

Authors: H. M. Takematsu, B. R. De Camargo, E. F. Noronha

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The use of hydrolyzing enzymes for degradation of lignocellulosic biomass is of great concern for the production of second generation ethanol. Although many hollocelulases have already been described in the literature, much more has to be discovered. Therefore, the aim of this study to evaluate hollocelulase production of P. polonicum grown in liquid media containing sugarcane bagasse as the carbon source. From a collection of twenty fungi isolated from Cerrado biome soil, P. polonicum was molecular identified by sequencing of ITS4, βtubulin and Calmodulin genes, and has been chosen to be further investigated regarding its potential production of hydrolyzing enzymes. Spore suspension (1x10-6 ml-1) solution was inoculated in sterilized minimal liquid medium containing 0,5%(w/v) of non-pretreated sugarcane bagasse as the carbon source, and incubated in shaker incubator at 28°C and 120 rpm. The supernatant obtained, was subjected to enzymatic assays to analyze xylanase, mannanase, pectinase and endoglucanase activities. Xylanase activity showed better results (67,36 UI/mg). Xylanases bands were indicated by zymogram and SDS-PAGE, and one of them was partially purified and characterized. It showed maximum activity at 50 °C, was thermostable for 72h at 40°C, and pH5.0 was the optimum observed. This study presents P. polonicum as an interesting source of hollocelulases, especially xylanase, for lignocellulose bio-conversion processes with commercial use.

Keywords: sugarcane bagasse, Cerrado biome , hollocelulase, lignocellulosic biomass

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283 Revolutionizing RNA Extraction: A Unified, Sustainable, and Rapid Protocol for High-Quality Isolation from Diverse Tissues

Authors: Ying Qi Chan, Chunyu Li, Xu Rou Yoyo Ma, Yaya Li, Saber Khederzadeh

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In the ever-evolving landscape of genome extraction protocols, the existing methodologies grapple with issues ranging from sub-optimal yields and compromised quality to time-intensive procedures and reliance on hazardous reagents, often necessitating substantial tissue quantities. This predicament is particularly challenging for scientists in developing countries, where resources are limited. Our investigation presents a protocol for the efficient extraction of high-yield RNA from various tissues such as muscle, insect, and plant samples. Noteworthy for its advantages, our protocol stands out as the safest, swiftest (completed in just 38 minutes), most cost-effective (coming in at a mere US$0.017), and highly efficient method in comparison to existing protocols. Notably, our method avoids the use of hazardous or toxic chemicals such as chloroform and phenol and enzymatic agents like RNase and Proteinase K. Our RNA extraction protocol has demonstrated clear advantages over other methods, including commercial kits, in terms of yield. This nucleic acid extraction protocol is more environmentally and research-friendly, suitable for a range of tissues, even in tiny volumes, hence facilitating various genetic diagnosis and researches across the globe.

Keywords: RNA extraction, rapid protocol, universal method, diverse tissues

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282 The Effect of Extrusion Processing on Solubility and Molecular Weight of Water-Soluble Arabinoxylan

Authors: Abdulmannan Fadel

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Arabinoxylan is a non-starch polysaccharide (NSP), which is one of the most important polysaccharides contained within cereal grains. Wheat endosperm pentosan and rice bran contain a significant amount of arabinoxylan (7% in rice bran and 10-12% in wheat endosperm pentosan). Several methods have been used for arabinoxylan extraction with varying degrees of success e.g. enzymatic and alkaline treatment. Yet, the use of extrusion alone as a pre-treatment to increase the yield and reduce the molecular weight in wheat endosperm pentosan and rice bran has not been investigated. The samples (wheat pentosan and rice bran) were extruded using a Twin-screw extruder at a range of screw speeds (80 and 160 rpm) and barrel temperatures range (80 to 140°C) with a throughput of 30 Kg hr-1 and moisture content of 25%. Arabinoxylans were extracted with water and the extraction yield and molecular weight was determined using size exclusion high-pressure liquid chromatography system. It was found that increasing screw speed from 80 rpm to 160 rpm, did not effect the extraction yield (p < 0.05) of arabinoxylan from either the wheat endosperm pentosan or the rice bran. However, the molecular weight of the extracted arabinoxylans from pentosan was found to decrease with increasing screw speed in wheat endosperm pentosan. These low molecular weight arabinoxylans have been suggested as immunomodulators.

Keywords: arabinoxylans, extrusion, wheat endosperm pentosan, rice bran

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281 Analysis of Histamine Content in Selected Food Products from the Serbian Market

Authors: Brizita Djordjevic, Bojana Vidovic, Milica Zrnic, Uros Cakar, Ivan Stankovic, Davor Korcok, Sladjana Sobajic

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Histamine is a biogenic amine, which is formed by enzymatic decarboxylation from the amino acid histidine. It can be found in foods such as fish and fish products, meat and fermented meat products, cheese, wine and beer. The presence of histamine in these foods can indicate microbiological spoilage or poor manufacturing processes. The consumption of food containing large amounts of histamine can have toxicological consequences. In 62 food products (31 canned fish products, 19 wines and 12 cheeses) from the market of Serbia the content of histamine was determined using enzyme-linked immunosorbent assay (ELISA) test kit according to the manufacturer's instructions (Immunolab GmbH, Kassel, Germany). The detection limits of this assay were 20 µg/kg for fish and cheese and 4 µg/L for wine. The concentration of histamine varied between 0.16-207 mg/kg in canned fish products, 0.03-1.47 mg/kg in cheeses and 0.01- 0.18 mg/L in wines. In all analyzed canned fish products the results obtained for the histamine were below the limits set by European and national legislation, so they can be considered acceptable and safe for the health consumers. The levels of histamine in analyzed cheeses and wines were very low and did not pose safety concerns.

Keywords: cheese, enzyme-linked immunosorbent assay, histamine, fish products, wine

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280 Dynamic Fault Diagnosis for Semi-Batch Reactor Under Closed-Loop Control via Independent RBFNN

Authors: Abdelkarim M. Ertiame, D. W. Yu, D. L. Yu, J. B. Gomm

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In this paper, a new robust fault detection and isolation (FDI) scheme is developed to monitor a multivariable nonlinear chemical process called the Chylla-Haase polymerization reactor when it is under the cascade PI control. The scheme employs a radial basis function neural network (RBFNN) in an independent mode to model the process dynamics and using the weighted sum-squared prediction error as the residual. The recursive orthogonal Least Squares algorithm (ROLS) is employed to train the model to overcome the training difficulty of the independent mode of the network. Then, another RBFNN is used as a fault classifier to isolate faults from different features involved in the residual vector. The several actuator and sensor faults are simulated in a nonlinear simulation of the reactor in Simulink. The scheme is used to detect and isolate the faults on-line. The simulation results show the effectiveness of the scheme even the process is subjected to disturbances and uncertainties including significant changes in the monomer feed rate, fouling factor, impurity factor, ambient temperature and measurement noise. The simulation results are presented to illustrate the effectiveness and robustness of the proposed method.

Keywords: Robust fault detection, cascade control, independent RBF model, RBF neural networks, Chylla-Haase reactor, FDI under closed-loop control

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279 Biochemical and Molecular Analysis of Staphylococcus aureus Various Isolates from Different Places

Authors: Kiran Fatima, Kashif Ali

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Staphylococcus aureus is an opportunistic human as well as animal pathogen that causes a variety of diseases. A total of 70 staphylococci isolates were obtained from soil, water, yogurt, and clinical samples. The likely staphylococci clinical isolates were identified phenotypically by different biochemical tests. Molecular identification was done by PCR using species-specific 16S rRNA primer pairs, and finally, 50 isolates were found to be positive as Staphylococcus aureus, sciuri, xylous and cohnii. Screened isolates were further analyzed by several microbiological diagnostics tests, including gram staining, coagulase, capsule, hemolysis, fermentation of glucose, lactose, maltose, and sucrose tests enzymatic reactions. It was found that 78%, 81%, and 51% of isolates were positive for gelatin hydrolysis, protease, and lipase activities, respectively. Antibiogram analysis of isolated Staphylococcus aureus strains with respect to different antimicrobial agents revealed resistance patterns ranging from 57 to 96%. Our study also shows 70% of strains to be MRSA, 54.3% as VRSA, and 54.3% as both MRSA and VRSA. All the identified isolates were subjected to detection of mecA, nuc, and hlb genes, and 70%, 84%, and 40% were found to harbour mecA, nuc, and hlb genes, respectively. The current investigation is highly important and informative for the high-level multidrug-resistant Staphylococcus aureus infections inclusive also of methicillin and vancomycin.

Keywords: MRSA, VRSA, mecA, MSSA

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278 Sardine Oil as a Source of Lipid in the Diet of Giant Freshwater Prawn (Macrobrachium rosenbergii)

Authors: A. T. Ramachandra Naik, H. Shivananda Murthy, H. n. Anjanayappa

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The freshwater prawn, Macrobrachium rosenbergii is a more popular crustacean cultured widely in monoculture system in India. It has got high nutritional value in the human diet. Hence, understanding its enzymatic and body composition is important in order to judge its flesh quality. Fish oil specially derived from Indian oil sardine is a good source of highly unsaturated fatty acid and lipid source in fish/prawn diet. A 35% crude protein diet with graded levels of Sardine oil as a source of fat was incorporated at four levels viz, 2.07, 4.07, 6.07 and 8.07% maintaining a total lipid level of feed at 8.11, 10.24, 12.28 and 14.33% respectively. Diet without sardine oil (6.05% total lipid) was served as basal treatment. The giant freshwater prawn, Macrobrachium rosenbergii was used as test animal and the experiment was lost for 112 days. Significantly, higher gain in weight of prawn was recorded in the treatment with 6.07% sardine oil incorporation followed by higher specific growth rate, food conversion rate and protein efficiency ratio. The 8.07% sardine oil diet produced the highest RNA: DNA ratio in the prawn muscle. Digestive enzyme analyses in the digestive tract and mid-gut gland showed the greatest activity in prawns fed the 8.07% diet.

Keywords: digestive enzyme, fish diet, Macrobrachium rosenbergii, sardine oil

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277 Pre-Drying Effects on the Quality of Frying Oil

Authors: Hasan Yalcin, Tugba Dursun Capar

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Deep-fat frying causes desirable as well as undesirable changes in oil and potato, and changes the quality of the oil by hydrolysis, oxidation, and polymerization. The main objective of the present study was to investigate the pre-drying effects on the quality of both frying oil and potatoes. Prior to frying, potato slices (10 mm x10 mm x 30 mm) were air- dried at 60°C for 15, 30, 45, 60, 90, and 120 mins., respectively. Potato slices without the pre-drying treatment were considered as the control variable. Potato slices were fried in sunflower oil at 180°C for 5, 10, and 13 mins. The deep-frying experiments were repeated five times using the new potato slices in the same oil without oil replenishment. Samples of the fresh oil, together with those sampled at the end of successive frying operations (1th, 3th and 5th) were removed and analysed. Moisture content, colour and oil intake of the potato and colour, peroxide value (PV), free fatty acid (FFA), fatty acid composition and viscosity of the used oil were evaluated. The effect of frying time was also examined. Results show that pre-drying treatment had a significant effect on physicochemical properties and colour parameters of potato slices and frying oil. Pre-drying considerably decreased the oil absorption. The lowest oil absorption was found for the treatment that was pre-dried for 120, and fried for 5 min. The FFA levels decreased permanently for each pre-treatment throughout the frying period. All the pre-drying treatments had reached their maximum levels of FFA by the end of the frying procedures. The PV of the control and 60 min pre-dried sample decreased after the third frying. However, the PV of other samples increased constantly throughout the frying periods. Lastly, pre-drying did not affect the fatty acid composition of frying oil considerably when compared against previously unused oil.

Keywords: air-drying, deep-fat frying, moisture content oil uptake, quality

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276 Laboratory Investigation of Fly Ash Based Geopolymer Stabilized Recycled Asphalt Pavement as a Base Material

Authors: Menglim Hoy, Suksun Horpibulsuk, Arul Arulrajah

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The results of laboratory investigation of recycled asphalt pavement (RAP) – fly ash (FA) based geopolymer as a base material is presented in this paper. An alkaline activator, the mixture of NaOH and Na₂SiO₃, is used to synthesis RAP-FA based geopolymer. RAP-FA with water (RAP-FA blend) prepared as a control material. The strength develops and the strength against wet-dry was determined by the unconfined compression strength (UCS) test, then the microstructural properties were examined by scanning electron microscopy (SEM) and X-ray Diffraction (XRD) analysis. The toxicity characteristic leaching procedure (TCLP) test is conducted to measure its leachability of heavy metal. The results show both the RAP-FA blend and geopolymer can be used as a base course as its UCS values meet the minimum strength requirement specified by the Department of Highway, Thailand. The durability test results show the UCS of these materials increases with increasing the number of wet-dry cycles, reaching its peak at six wet-dry cycles. The XRD and SEM analyses indicate strength development of the RAP-FA blend occurs due to chemical reaction between a high Calcium in RAP with a high Silica and Alumina in FA led to producing calcium aluminate hydrate formation. The strength development of the RAP-FA geopolymer occurred resulted from the polymerization reaction. The TCLP results demonstrate there is no environmental risk of these stabilized materials. Furthermore, FA based geopolymer can reduce the leachability of heavy metal in the RAP-FA blend.

Keywords: recycled asphalt pavement, geopolymer, heavy metal, microstructure

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275 Can We Meet the New Challenges of NonIsocyanates Polyurethanes (NIPU) towards NIPU Foams?

Authors: Adrien Cornille, Marine Blain, Bernard Boutevin, Sylvain Caillol

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Generally, linear polyurethanes (PUs) are obtained by the reaction between an oligomeric diol, a short diol as chain extender and a diisocyanate. However the use of diisocyanate should be avoided since they are generally very harmful for human health. Therefore the synthesis of NIPUs (non isocyanate PUs) from step growth polymerization of dicyclocarbonates and diamines should be favoured. This method is particularly interesting since no hazardous isocyanates are used. Thus, this reaction, extensively studied by Endo et al. is currently gaining a lot of attention as a substitution route for the synthesis of NIPUs, both from industrial and academic community. However, the reactivity of reaction between amine and cyclic carbonate is a major scientific issue, since cyclic carbonates are poorly reactive. Thus, our team developed several synthetic ways for the synthesis of various di-cyclic carbonates based on C5-, C6- and dithio- cyclic carbonates, from different biobased raw materials (glycerin isosorbide, vegetable oils…). These monomers were used to synthesize NIPUs with various mechanical and thermal properties for various applications. We studied the reactivity of reaction with various catalysts and find optimized conditions for room temperature reaction. We also studied the radical copolymerization of cyclic carbonate monomers in styrene-acrylate copolymers for coating applications. We also succeeded in the elaboration of biobased NIPU flexible foams. To the best of our knowledge, there is no report in literature on the preparation of non-isocyanate polyurethane foams.

Keywords: foam, nonisocyanate polyurethane, cyclic carbonate, blowing agent, scanning electron microscopy

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274 VII Phytochemistry UNIT-IV Glycoside

Authors: Magy Magdy Danial Riad

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Introduction: Glycosides: Enzymatic and hydrolysis reactions of glycosides, mechanism of action, SAR, therapeutic uses and toxicity of glycosides. Cardiac glycosides of digitalis, bufa and squill. Structure of salicin, hesperidin and rutin. Glycosides are certain molecules in which a sugar part is bound to some other part. Glycosides play numerous important roles in living organisms. Formally, a glycoside is any molecule in which a sugar group is bonded through its anomeric carbon to another group and form glycosidic bonds via an O-glycosidic bond or an S-glycosidic bond; glycosides involving the latter are also called thioglycosides. The purpose: the addition of sugar be bonded to a non-sugar for the molecule to qualify as a glycoside, The sugar group is then known as the glycone and the non-sugar group as the aglycone or genin part of the glycoside. The glycone can consist of a single sugar group (monosaccharide) or several sugar groups (oligosaccharide). The glycone and aglycone portions can be chemically separated by hydrolysis in the presence of acid. Methods: There are also numerous enzymes that can form and break glycosidic bonds. Results: The most important cleavage enzymes are the glycoside hydrolases, and the most important synthetic enzymes in nature are glycosyltransferases. Mutant enzymes termed glycosynthases have been developed that can form glycosidic bonds. Conclusions: There are a great many ways to chemically synthesize glycosidic bonds.

Keywords: glycosides, bufa, squill, thioglycosides

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273 Modifiable Poly Methacrylic Acid-Co-Acrylonitrile Microgels Fabricated with Cu and Co Nanoparticles for Simultaneous Catalytic Reduction of Multiple Compounds

Authors: Muhammad Ajmal, Muhammad Siddiq, Nurettin Sahiner

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We prepared poly(methacrylic acid-co-acrylonitrile) (p(MAc-co-AN)) microgels by inverse suspension polymerization, and converted the nitrile groups into amidoxime groups to obtain more hydrophilic amidoximated poly(methacrylic acid-co-acrylonitile) (amid-p(MAc-co-AN)) microgels. Amid-microgels were used as microreactors for in situ synthesis of copper and cobalt nanoparticles. Cu (II) and Co (II) ions were loaded into microgels from their aqueous metal salt solutions and then converted to corresponding metal nanoparticle (MNP) by treating the loaded metal ions with sodium borohydride (NaBH4). The characterization of the prepared microgels and microgel metal nanoparticle composites was carried out by SEM, TEM and TG analysis. The amounts of metal nanoparticles within microgels were estimated by AAS measurements by dissolving the MNP entrapped within microgels by concentrated HCl acid treatment. Catalytic performances of the prepared amid-p(MAc-co-AN)-M (M: Cu, Co) microgel composites were investigated by using them as catalyst for the degradation of cationic and anionic organic dyes such as eosin Y (EY), methylene blue (MB) and methyl Orange (MO), and for the reduction of nitro aromatic pollutants like 2-nitrophenol (2-NP) and 4-nitrophenol (4-NP) to their corresponding amino phenols. Here, we also report for the first time, the simultaneous degradation/reduction of MB, EY, and 4-NP by amid-p(MAc-co-AN)-Cu microgel composites. Different parameters affecting the reduction rates such as metal types, amount of catalysts, temperature and the amount of reducing agent were investigated.

Keywords: microgels, nanoparticles, catalyst, pollutants

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272 Effect of Thermal Pretreatment on Functional Properties of Chicken Protein Hydrolysate

Authors: Nutnicha Wongpadungkiat, Suwit Siriwatanayotin, Aluck Thipayarat, Punchira Vongsawasdi, Chotika Viriyarattanasak

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Chicken products are major export product of Thailand. With a dramatically increasing consumption of chicken product in the world, there are abundant wastes from chicken meat processing industry. Recently, much research in the development of value-added products from chicken meat industry has focused on the production of protein hydrolysate, utilized as food ingredients for human diet and animal feed. The present study aimed to determine the effect of thermal pre-treatment on functional properties of chicken protein hydrolysate. Chicken breasts were heated at 40, 60, 80 and 100ºC prior to hydrolysis by Alcalase at 60ºC, pH 8 for 4 hr. The hydrolysate was freeze-dried, and subsequently used for assessment of its functional properties molecular weight by gel electrophoresis (SDS-PAGE). The obtained results show that increasing the pre-treatment temperature increased oil holding capacity and emulsion stability while decreasing antioxidant activity and water holding capacity. The SDS-PAGE analysis showed the evidence of protein aggregation in the hydrolysate treated at the higher pre-treatment temperature. These results suggest the connection between molecular weight of the hydrolysate and its functional properties.

Keywords: chicken protein hydrolysate, enzymatic hydrolysis, thermal pretreatment, functional properties

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271 Fabrication of Electrospun Carbon Nanofibers-Reinforced Chitosan-Based Hydrogel for Environmental Applications

Authors: Badr M. Thamer

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The use of hydrogels as adsorbents for pollutants removal from wastewater is limited due to their high swelling properties and the difficulty in recovering them after the adsorption process. To overcome these problems, a new hydrogel nanocomposite based on chitosan-g-polyacrylic acid/oxidized electrospun carbon nanofibers (CT-g-PAA/O-ECNFs) was prepared by in-situ grafting polymerization process. The prepared hydrogel nanocomposite was used as a novel effective and highly reusable adsorbent for the removal of methylene blue (MB) from polluted water with low cost. The morphology and the structure of CT-g-PAA/O-ECNFs were investigated by numerous techniques. The effect of incorporating O-ECNFs on the swelling capability of the prepared hydrogel was explored in distillated water and MB solution at normal pH. The effect of parameters including the ratio of O-ECNFs, contact time, pH, initial concentration, and temperature on the adsorption process were explored. The adsorption isotherm and kinetic were studied by numerous non-linear models. The obtained results confirmed that the incorporation of O-ECNFs into the hydrogel network improved its ability towards MB dye removal with decreasing their swelling capacity. The adsorption process depends on the pH value of the dye solution. Additionally, the adsorption and kinetic results were fitted using the Freundlich isotherm model and pseudo second order model (PSO), respectively. Moreover, the new adsorbents can be recycled for at least five cycles keeping its adsorption capacity and can be easily recovered without loss in its initial weight.

Keywords: carbon nanofibers, hydrogels, nanocomposites, water treatment

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270 Genetic and Non-Genetic Factors Affecting the Response to Clopidogrel Therapy

Authors: Snezana Mugosa, Zoran Todorovic, Zoran Bukumiric, Ivan Radosavljevic, Natasa Djordjevic

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Introduction: Various studies have shown that the frequency of clopidogrel resistance ranges from 4-40%. The aim of this study was to provide in depth analysis of genetic and non-genetic factors that influence clopidogrel resistance in cardiology patients. Methods: We have conducted a prospective study in 200 hospitalized patients hospitalized at Cardiology Centre of the Clinical Centre of Montenegro. CYP2C19 genetic testing was conducted, and the PREDICT score was calculated in 102 out of 200 patients treated with clopidogrel in order to determine the influence of genetic and non-genetic factors on outcomes of interest. Adverse cardiovascular events and adverse reactions to clopidogrel were assessed during 12 months follow up period. Results: PREDICT score and CYP2C19 enzymatic activity were found to be statistically significant predictors of expressing lack of therapeutic efficacy of clopidogrel by multivariate logistic regression, without multicollinearity or interaction between the predictors (p = 0.002 and 0.009, respectively). Conclusions: Pharmacogenetics analyses that were done in the Montenegrin population of patients for the first time suggest that these analyses can predict patient response to the certain therapy. Stepwise approach could be used in assessing the clopidogrel resistance in cardiology patients, combining the PREDICT score, platelet aggregation test, and genetic testing for CYP2C19 polymorphism.

Keywords: clopidogrel, pharmacogenetics, pharmacotherapy, PREDICT score

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269 The Effect of Artesunate on Myeloperoxidase Activity of Human Polymorphonuclear Neutrophil

Authors: J. B. Minari, O. B. Oloyede, A. A. Odutuga

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Myeloperoxidase is the most abundant enzyme found in the polymorphonuclear neutrophil and is known to play a central role in the host defense system of the leukocyte. The enzyme has been reported to interact with some drugs to generate free radical which inhibits its activity. This study investigated the effects of artesunate on the activity of the enzyme and the subsequent effect on the host immune system. In investigating the effects of the drugs on myeloperoxidase, the influence of concentration, pH, partition ratio estimation and kinetics of inhibition were studied. This study showed that artesunate is concentration-dependent inhibitor of myeloperoxidase with an IC50 of 0.078mM. Partition ratio estimation showed that 60 enzymatic turnover cycles are required for complete inhibition of myeloperoxidase in the presence of artesunate. The influence of pH on the effect of artesunate on the enzyme showed least activity of myeloperoxidase at physiological pH. The kinetic inhibition studies showed that artesunate caused a competitive inhibition with an increase in the Km value from 0.12mM to 0.26mM and no effect on the Vmax value. The Ki value was estimated to be 2.5mM. The results obtained from this study show that artesunate is a potent inhibitor of myeloperoxidase and it is capable of inactivating the enzyme. It is considered that the inhibition of myeloperoxidase in the presence of artesunate as revealed in this study may partly explain the impairment of polymorphonuclear neutrophil and consequent reduction of the strength of the host defense system against secondary infections.

Keywords: myeloperoxidase, artesunate, inhibition, nuetrophill

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268 Evaluation of Chromium Fortified-Parboiled Rice Coated with Herbal Extracts: Resistant Starch, and Glycemic Index

Authors: Wisnu Adi Yulianto, Chatarina Lilis Suryani, Mamilisti Susiati, Hendy Indra Permana

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Parboiled rice was developed to produce rice that has low glycemic index, especially for diabetics. Yet, parboiled rice is not enough because diabetics also lack of chromium. The sign of chromium (Cr) deficiency in diabetics is impaired glucose tolerance. Cr fortification was done for increasing Cr content in rice. Naturally-occurring compounds that have been proven to improve insulin sensitivity include Cr and polyphenol found in cinnamon, pandan and bay leaf. This research aimed to evaluate content of resistant starch and glycemic index of Cr - fortified - parboiled rice (Cr-PR) coated with herbal extracts. Variety of unhulled rice and forticant used in the experiment were Ciherang and CrCl3, respectively. Three herbal extracts used were cinnamon, pandan and bay leaf. Each concentration of herbal extracts in the amount of 3%, 6%, and 9% were added in the coating substance to coat Cr-PR. Resistant starch (RS) content was determined by enzymatic process through glucooxydase method. Testing of the GI was conducted on 18 non-diabetic volunteers. RS content of Cr-PR coated with herbal extracts ranged between 8.27 – 8.84 % (dry weight). Cr-PR coated with all herbal extracts of 3% concentration had higher RS content than the ones with herbal extracts of 6% and 9% concentration (P <0.05). Value of the rice GI ranged 29 - 40. The lowest GI (29-30) was attained by the rice coated with enrichment of 6-9% cinnamon extract.

Keywords: coating, Cr-fortified-parboiled rice, glycemic index, herbal extracts, resistant starch

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267 Functional Cell Surface Display Using Ice Nucleation Protein from Erwina ananas on Escherischia coli

Authors: Mei Yuin Joanne Wee, Rosli Md. Illias

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Cell surface display is the expression of a protein with an anchoring motif on the surface of the cell. This approach offers advantages when used in bioconversion in terms of easier purification steps and more efficient enzymatic reaction. A surface display system using ice nucleation protein (InaA) from Erwina ananas as an anchoring motif has been constructed to display xylanase (xyl) on the surface of Escherischia coli. The InaA was truncated so that it is made up of the N- and C-terminal domain (INPANC-xyl) and it has successfully directed xylanase to the surface of the cell. A study was also done on xylanase fused to two other ice nucleation proteins, InaK (INPKNC-xyl) and InaZ (INPZNC-xyl) from Pseudomonas syringae KCTC 1832 and Pseudomonas syringae S203 respectively. Surface localization of the fusion protein was verified using SDS-PAGE and Western blot on the cell fractions and all anchoring motifs were successfully displayed on the outer membrane of E. coli. Upon comparison, whole-cell activity of INPANC-xyl was more than six and five times higher than INPKNC-xyl and INPZNC-xyl respectively. Furthermore, the expression of INPANC-xyl on the surface of E. coli did not inhibit the growth of the cell. This is the first report of surface display system using ice nucleation protein, InaA from E. ananas. From this study, this anchoring motif offers an attractive alternative to the current surface display systems.

Keywords: cell surface display, Escherischia coli, ice nucleation protein, xylanase

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266 Preparation and Characterisation of Electrospun Extracted β-Chitosan/Poly(Vinyl Alcohol) Blend Nanofibers for Tissue Engineering

Authors: E. Roshan Ara Begum, K. Bhavani, K. Subachitra, C. Kirthika, R. Shenbagarathai

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In recent years, there has been a growing concern for the production of chitosan blend nanofibrous scaffold for its favorable physicochemical properties which mimic the native extracellular matrix (ECM) both morphologically and chemically. Therefore, this study focused on production of β-chitosan(β-Cts) and Poly(vinyl alcohol)(PVA) blend nanofibrous scaffold by electrospinning. β-Cts was extracted from the squid pen waste of locally available squid variety Loligo duvauceli (Indian Squid). To the best of our knowledge, there are no reports on nanofibers preparation from the extracted β-Cts. Both the β-Cts and PVA polymers were mixed in two different proportions (30:70 and 40:60 respectively. The electrospun nanofibrous scaffolds were characterized by SEM, swelling property, in vitro enzymatic degradation, and hemo, biocompatibility properties. β-Cts/PVA nanofibers scaffolds had an average fiber diameter of 120 to 550nm.Among the two different β-Cts/PVA blends nanofibers the β-Cts/PVA (40:60) blend fibers demonstrated favourable tissue engineering properties. The β-Cts/PVA (40:60) blend nanofibers exhibited a swelling ratio of 36 ± 2.5% with mass loss percentage of 20 ± 2.71% after 4 weeks of degradation. It has exhibited good hemocompatible properties. HEK-293(Human Embryonic Kidney) cells lines were able to adhere and proliferate well in the β-Cts/PVA blends nanofibers. All these results indicated that electrospun β-Cts/PVA blends nanofibers are a suitable scaffold to be used for tissue engineering purposes.

Keywords: β-chitosan, electrospinning, nanofibers, poly(vinyl alcohol) (PVA)

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265 The Investigation of Enzymatic Activity in the Soils Under the Impact of Metallurgical Industrial Activity in Lori Marz, Armenia

Authors: T. H. Derdzyan, K. A. Ghazaryan, G. A. Gevorgyan

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Beta-glucosidase, chitinase, leucine-aminopeptidase, acid phosphomonoestearse and acetate-esterase enzyme activities in the soils under the impact of metallurgical industrial activity in Lori marz (district) were investigated. The results of the study showed that the activities of the investigated enzymes in the soils decreased with increasing distance from the Shamlugh copper mine, the Chochkan tailings storage facility and the ore transportation road. Statistical analysis revealed that the activities of the enzymes were positively correlated (significant) to each other according to the observation sites which indicated that enzyme activities were affected by the same anthropogenic factor. The investigations showed that the soils were polluted with heavy metals (Cu, Pb, As, Co, Ni, Zn) due to copper mining activity in this territory. The results of Pearson correlation analysis revealed a significant negative correlation between heavy metal pollution degree (Nemerow integrated pollution index) and soil enzyme activity. All of this indicated that copper mining activity in this territory causing the heavy metal pollution of the soils resulted in the inhabitation of the activities of the enzymes which are considered as biological catalysts to decompose organic materials and facilitate the cycling of nutrients.

Keywords: Armenia, metallurgical industrial activity, heavy metal pollutionl, soil enzyme activity

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