Search results for: angiotensin converting enzyme
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 1154

Search results for: angiotensin converting enzyme

734 Biohydrogen Production from Starch Residues

Authors: Francielo Vendruscolo

Abstract:

This review summarizes the potential of starch agroindustrial residues as substrate for biohydrogen production. Types of potential starch agroindustrial residues, recent developments and bio-processing conditions for biohydrogen production will be discussed. Biohydrogen is a clean energy source with great potential to be an alternative fuel, because it releases energy explosively in heat engines or generates electricity in fuel cells producing water as only by-product. Anaerobic hydrogen fermentation or dark fermentation seems to be more favorable, since hydrogen is yielded at high rates and various organic waste enriched with carbohydrates as substrate result in low cost for hydrogen production. Abundant biomass from various industries could be source for biohydrogen production where combination of waste treatment and energy production would be an advantage. Carbohydrate-rich nitrogen-deficient solid wastes such as starch residues can be used for hydrogen production by using suitable bioprocess technologies. Alternatively, converting biomass into gaseous fuels, such as biohydrogen is possibly the most efficient way to use these agroindustrial residues.

Keywords: biofuel, dark fermentation, starch residues, food waste

Procedia PDF Downloads 371
733 Significance of Molecular Autophagic Pathway in Gaucher Disease Pathology

Authors: Ozlem Oral, Emre Taskin, Aysel Yuce, Serap Dokmeci, Devrim Gozuacik

Abstract:

Autophagy is an evolutionary conserved lysosome-dependent catabolic pathway, responsible for the degradation of long-lived proteins, abnormal aggregates and damaged organelles which cannot be degraded by the ubiquitin-proteasome system. Lysosomes degrade the substrates through the activity of lysosomal hydrolases and lysosomal membrane-bound proteins. Mutations in the coding region of these proteins cause malfunctional lysosomes, which contributes to the pathogenesis of lysosomal storage diseases. Gaucher disease is a lysosomal storage disease resulting from the mutation of a lysosomal membrane-associated glycoprotein called glucocerebrosidase and its cofactor saposin C. The disease leads to intracellular accumulation of glucosylceramide and other glycolipids. Because of the essential role of lysosomes in autophagic degradation, Gaucher disease may directly be linked to this pathway. In this study, we investigated the expression of autophagy and/or lysosome-related genes and proteins in fibroblast cells isolated from patients with different mutations. We carried out confocal microscopy analysis and examined autophagic flux by utilizing the differential pH sensitivities of RFP and GFP in mRFP-GFP-LC3 probe. We also evaluated lysosomal pH by active lysosome staining and lysosomal enzyme activity. Beside lysosomes, we also performed proteasomal activity and cell death analysis in patient samples. Our data showed significant attenuation in the expression of key autophagy-related genes and accumulation of their proteins in mutant cells. We found decreased the ability of autophagosomes to fuse with lysosomes, associated with elevated lysosomal pH and reduced lysosomal enzyme activity. Proteasomal degradation and cell death analysis showed reduced proteolytic activity of the proteasome, which consequently leads to increased susceptibility to cell death. Our data indicate that the major degradation pathways are affected by multifunctional lysosomes in mutant patient cells and may underlie in the mechanism of clinical severity of Gaucher patients. (This project is supported by TUBITAK-3501-National Young Researchers Career Development Program, Project No: 112T130).

Keywords: autophagy, Gaucher's disease, glucocerebrosidase, mutant fibroblasts

Procedia PDF Downloads 304
732 Exo-III Assisted Amplification Strategy through Target Recycling of Hg²⁺ Detection in Water: A GNP Based Label-Free Colorimetry Employing T-Rich Hairpin-Loop Metallobase

Authors: Abdul Ghaffar Memon, Xiao Hong Zhou, Yunpeng Xing, Ruoyu Wang, Miao He

Abstract:

Due to deleterious environmental and health effects of the Hg²⁺ ions, various online, detection methods apart from the traditional analytical tools have been developed by researchers. Biosensors especially, label, label-free, colorimetric and optical sensors have advanced with sensitive detection. However, there remains a gap of ultrasensitive quantification as noise interact significantly especially in the AuNP based label-free colorimetry. This study reported an amplification strategy using Exo-III enzyme for target recycling of Hg²⁺ ions in a T-rich hairpin loop metallobase label-free colorimetric nanosensor with an improved sensitivity using unmodified gold nanoparticles (uGNPs) as an indicator. The two T-rich metallobase hairpin loop structures as 5’- CTT TCA TAC ATA GAA AAT GTA TGT TTG -3 (HgS1), and 5’- GGC TTT GAG CGC TAA GAA A TA GCG CTC TTT G -3’ (HgS2) were tested in the study. The thermodynamic properties of HgS1 and HgS2 were calculated using online tools (http://biophysics.idtdna.com/cgi-bin/meltCalculator.cgi). The lab scale synthesized uGNPs were utilized in the analysis. The DNA sequence had T-rich bases on both tails end, which in the presence of Hg²⁺ forms a T-Hg²⁺-T mismatch, promoting the formation of dsDNA. Later, the Exo-III incubation enable the enzyme to cleave stepwise mononucleotides from the 3’ end until the structure become single-stranded. These ssDNA fragments then adsorb on the surface of AuNPs in their presence and protect AuNPs from the induced salt aggregation. The visible change in color from blue (aggregation stage in the absence of Hg²⁺) and pink (dispersion state in the presence of Hg²⁺ and adsorption of ssDNA fragments) can be observed and analyzed through UV spectrometry. An ultrasensitive quantitative nanosensor employing Exo-III assisted target recycling of mercury ions through label-free colorimetry with nanomolar detection using uGNPs have been achieved and is further under the optimization to achieve picomolar range by avoiding the influence of the environmental matrix. The proposed strategy will supplement in the direction of uGNP based ultrasensitive, rapid, onsite, label-free colorimetric detection.

Keywords: colorimetric, Exo-III, gold nanoparticles, Hg²⁺ detection, label-free, signal amplification

Procedia PDF Downloads 289
731 Clostridium thermocellum DBT-IOC-C19, A Potential CBP Isolate for Ethanol Production

Authors: Nisha Singh, Munish Puri, Collin Barrow, Deepak Tuli, Anshu S. Mathur

Abstract:

The biological conversion of lignocellulosic biomass to ethanol is a promising strategy to solve the present global crisis of exhausting fossil fuels. The existing bioethanol production technologies have cost constraints due to the involvement of mandate pretreatment and extensive enzyme production steps. A unique process configuration known as consolidated bioprocessing (CBP) is believed to be a potential cost-effective process due to its efficient integration of enzyme production, saccharification, and fermentation into one step. Due to several favorable reasons like single step conversion, no need of adding exogenous enzymes and facilitated product recovery, CBP has gained the attention of researchers worldwide. However, there are several technical and economic barriers which need to be overcome for making consolidated bioprocessing a commercially viable process. Finding a natural candidate CBP organism is critically important and thermophilic anaerobes are preferred microorganisms. The thermophilic anaerobes that can represent CBP mainly belong to genus Clostridium, Caldicellulosiruptor, Thermoanaerobacter, Thermoanaero bacterium, and Geobacillus etc. Amongst them, Clostridium thermocellum has received increased attention as a high utility CBP candidate due to its highest growth rate on crystalline cellulose, the presence of highly efficient cellulosome system and ability to produce ethanol directly from cellulose. Recently with the availability of genetic and molecular tools aiding the metabolic engineering of Clostridium thermocellum have further facilitated the viability of commercial CBP process. With this view, we have specifically screened cellulolytic and xylanolytic thermophilic anaerobic ethanol producing bacteria, from unexplored hot spring/s in India. One of the isolates is a potential CBP organism identified as a new strain of Clostridium thermocellum. This strain has shown superior avicel and xylan degradation under unoptimized conditions compared to reported wild type strains of Clostridium thermocellum and produced more than 50 mM ethanol in 72 hours from 1 % avicel at 60°C. Besides, this strain shows good ethanol tolerance and growth on both hexose and pentose sugars. Hence, with further optimization this new strain could be developed as a potential CBP microbe.

Keywords: Clostridium thermocellum, consolidated bioprocessing, ethanol, thermophilic anaerobes

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730 Prevalence and Mechanisms of Antibiotic Resistance in Escherichia coli Isolated from Mastitic Dairy Cattle in Canada

Authors: Satwik Majumder, Dongyun Jung, Jennifer Ronholm, Saji George

Abstract:

Bovine mastitis is the most common infectious disease in dairy cattle, with major economic implications for the dairy industry worldwide. Continuous monitoring for the emergence of antimicrobial resistance (AMR) among bacterial isolates from dairy farms is vital not only for animal husbandry but also for public health. In this study, the prevalence of AMR in 113 Escherichia coli isolates from cases of bovine clinical mastitis in Canada was investigated. Kirby-Bauer disk diffusion test with 18 antibiotics and microdilution method with three heavy metals (copper, zinc, and silver) was performed to determine the antibiotic and heavy-metal susceptibility. Resistant strains were assessed for efflux and ß-lactamase activities besides assessing biofilm formation and hemolysis. Whole-genome sequences for each of the isolates were examined to detect the presence of genes corresponding to the observed AMR and virulence factors. Phenotypic analysis revealed that 32 isolates were resistant to one or more antibiotics, and 107 showed resistance against at least one heavy metal. Quinolones and silver were the most efficient against the tested isolates. Among the AMR isolates, AcrAB-TolC efflux activity and ß-lactamase enzyme activities were detected in 13 and 14 isolates, respectively. All isolates produced biofilm but with different capacities, and 33 isolates showed α-hemolysin activity. A positive correlation (Pearson r = +0.89) between efflux pump activity and quantity of biofilm was observed. Genes associated with aggregation, adhesion, cyclic di-GMP, quorum sensing were detected in the AMR isolates, corroborating phenotype observations. This investigation showed the prevalence of AMR in E. coli isolates from bovine clinical mastitis. The results also suggest the inadequacy of antimicrobials with a single mode of action to curtail AMR bacteria with multiple mechanisms of resistance and virulence factors. Therefore, it calls for combinatorial therapy for the effective management of AMR infections in dairy farms and combats its potential transmission to the food supply chain through milk and dairy products.

Keywords: antimicrobial resistance, E. coli, bovine mastitis, antibiotics, heavy-metals, efflux pump, ß-lactamase enzyme, biofilm, whole-genome sequencing

Procedia PDF Downloads 181
729 Caffeic Acid in Cosmetic Formulations: An Innovative Assessment

Authors: Caroline M. Spagnol, Vera L. B. Isaac, Marcos A. Corrêa, Hérida R. N. Salgado

Abstract:

Phenolic compounds are abundant in the Brazilian plant kingdom and they are part of a large and complex group of organic substances. Cinnamic acids are part of this group of organic compounds, and caffeic acid (CA) is one of its representatives. Antioxidants are compounds which act as free radical scavengers and, in other cases, such as metal chelators, both in the initiation stage and the propagation of oxidative process. The tyrosinase, polyphenol oxidase, is an enzyme that acts at various stages of melanin biosynthesis within the melanocytes and is considered a key molecule in this process. Some phenolic compounds exhibit inhibitory effects on melanogenesis by inhibiting the tyrosinase enzymatic activity and therefore has been the subject of studies. However, few studies have reported the effectiveness of these products and their safety. Objectives: To assess the inhibitory activity of tyrosinase, the antioxidant activity of CA and its cytotoxic potential. The method to evaluate the inhibitory activity of tyrosinase aims to assess the reduction transformation of L-dopa into dopaquinone reactions catalyzed by the enzyme. For evaluating the antioxidant activity was used the analytical methodology of DPPH radical inhibition. The cytotoxicity evaluation was carried out using the MTT method (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide), a colorimetric assay which determines the amount of insoluble violet crystals formed by the reduction of MTT in the mitochondria of living cells. Based on the results obtained during the study, CA has low activity as a depigmenting agent. However, it is a more potent antioxidant than ascorbic acid (AA), since a lower amount of CA is sufficient to inhibit 50% of DPPH radical. The results are promising since CA concentration that promoted 50% toxicity in HepG2 cells (IC50=781.8 μg/mL) is approximately 330 to 400 times greater than the concentration required to inhibit 50% of DPPH (IC50 DPPH= 2.39 μg/mL) and ABTS (IC50 ABTS= 1.96 μg/mL) radicals scavenging activity, respectively. The maximum concentration of caffeic acid tested (1140 mg /mL) did not reach 50% of cell death in HaCat cells. Thus, it was concluded that the caffeic acid does not cause toxicity in HepG2 and HaCat cells in the concentrations required to promote antioxidant activity in vitro, and it can be applied in topical products.

Keywords: caffeic acid, antioxidant, cytotoxicity, cosmetic

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728 Observation on Microbiological Profile of Type2 Diabetic Foot Ulcer and Its Antimicrobial Sensitivity Pattern in a Tertiary Care Hospital in Eastern India

Authors: Pampita Chakraborty, Sukumar Mukherjee

Abstract:

Diabetes Mellitus (DM) is commonly encountered metabolic disorder in clinical practice. An estimated 25 percent of DM patients develop foot problems. Foot ulceration and infection are one of the major causes of morbidity, hospitalization or even amputation. Objective: To isolate and identify bacterial pathogens in Diabetic Foot Ulcer (DFU) and to observe its antimicrobial sensitivity pattern. Methodology: A prospective study was conducted for a period of 9 months at the Department of Microbiology, GD Hospital & Diabetes Institute, Kolkata. 75 DFU patients were recruited in the study. Specimens for microbiological studies obtained from ulcer base were examined as gram stained smear and was cultured aerobically on Nutrient agar, Blood agar and MacConkey agar plates. Antimicrobial sensitivity test was performed by disc diffusion techniques according to CLSI guidelines. Result: In this study out of 75cases, 73% (55/75) were male and 27% (20/75) were females with mean (SD) age of 51.11(±10) years. Out of 75 pus cultures, 63(84%) showed growth of microorganism making total of 81 bacterial isolates with 71.42% of monomicrobial infection and 28.57% of polymicrobial infection. Out of 81 isolates 53(65.43%) were gram negative and 21(25.92%) were gram positive. E.coli was relatively common isolate 21(26%) followed by Staphylococcus aureus 15(18.5%), Klebsiella pneumonia 14(17.28%), Pseudomonas aeruginosa 12 (14.81%), Proteus spp. 3 (3.70%), and Enterococcus faecalis 6 (7.40%). 75% of Gram-negative microorganism were extended Beta-lactamase enzyme (ESBL) producer and around 20 % of Klebsiella and Proteus spp. were carbapenemase enzyme producer. Among Gram positive, around 50% of S.aureus was MRSA, sensitive only to Vancomycin, Teicoplanin & Linezolid. Conclusion: More prevalence of monomicrobial gram-negative bacteria than gram-positive bacteria in DFU was observed. This study emphasizes that Beta-Lactam group of antibiotics should not be the empirical treatment of choice for Gram-negative isolates; instead alternatives like Carbapenems, Amikacin could be a better option. On the other hand, Vancomycin and Linezolid are preferred for most of the infection with gram-positive aerobes. Continuous surveillance of resistant bacteria is required for empiric therapy.

Keywords: antibiotic resistant, antimicrobial susceptibility, diabetic foot ulcer, surveillance

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727 The Effect of Mean Pressure on the Performance of a Low-Grade Heat-Driven Thermoacoustic Cooler

Authors: Irna Farikhah

Abstract:

Converting low-grade waste heat into useful energy such as sound energy which can then be used to generate acoustic power in a thermoacoustic engine has become an attracting issue for researchers. The generated power in thermoacoustic engine can be used for driving a thermoacoustic cooler when they are installed in a tube. This cooler system can be called as a heat-driven thermoacoustic cooler. In this study, low heating temperature of the engine is discussed. In addition, having high efficiency of the whole cooler is also essential. To design a thermoacoustic cooler having high efficiency with using low-grade waste heat for the engine, the effect of mean pressure is investigated. By increasing the mean pressure, the heating temperature to generate acoustic power can be decreased from 557 °C to 300 °C. Moreover, the efficiency of the engine and cooler regenerators attain 67% and 47% of the upper limit values, respectively and 49% of the acoustical work generated by the engine regenerator is utilized in the cooler regenerator. As a result, the efficiency of the whole cooler becomes 15% of the upper limit value.

Keywords: cooler, mean pressure, performance, thermoacoustic

Procedia PDF Downloads 234
726 Investigation of Dry-Blanching and Freezing Methods of Fruits

Authors: Epameinondas Xanthakis, Erik Kaunisto, Alain Le-Bail, Lilia Ahrné

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Fruits and vegetables are characterized as perishable food matrices due to their short shelf life as several deterioration mechanisms are being involved. Prior to the common preservation methods like freezing or canning, fruits and vegetables are being blanched in order to inactivate deteriorative enzymes. Both conventional blanching pretreatments and conventional freezing methods hide drawbacks behind their beneficial impacts on the preservation of those matrices. Conventional blanching methods may require longer processing times, leaching of minerals and nutrients due to the contact with the warm water which in turn leads to effluent production with large BOD. An important issue of freezing technologies is the size of the formed ice crystals which is also critical for the final quality of the frozen food as it can cause irreversible damage to the cellular structure and subsequently to degrade the texture and the colour of the product. Herein, the developed microwave blanching methodology and the results regarding quality aspects and enzyme inactivation will be presented. Moreover, heat transfer phenomena, mass balance, temperature distribution, and enzyme inactivation (such as Pectin Methyl Esterase and Ascorbic Acid Oxidase) of our microwave blanching approach will be evaluated based on measurements and computer modelling. The present work is part of the COLDμWAVE project which aims to the development of an innovative environmentally sustainable process for blanching and freezing of fruits and vegetables with improved textural and nutritional quality. In this context, COLDµWAVE will develop tailored equipment for MW blanching of vegetables that has very high energy efficiency and no water consumption. Furthermore, the next steps of this project regarding the development of innovative pathways in MW assisted freezing to improve the quality of frozen vegetables, by exploring in depth previous results acquired by the authors, will be presented. The application of MW assisted freezing process on fruits and vegetables it is expected to lead to improved quality characteristics compared to the conventional freezing. Acknowledgments: COLDμWAVE has received funding from the European Union’s Horizon 2020 research and innovation programme under the Marie Sklodowska-Curie grand agreement No 660067.

Keywords: blanching, freezing, fruits, microwave blanching, microwave

Procedia PDF Downloads 239
725 Synthesis of Green Fuel Additive from Waste Bio-Glycerol

Authors: Ala’a H. Al-Muhtaseb, Farrukh Jamil, Lamya Al-Haj, Mohab Al-Hinai

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Bio-glycerol is considered as high boiling polar triol and immiscible with fossil fuel fractions due to which it is transformed into its respective ketals and acetals which help to improve the quality of diesel emitting less amount of aldehydes and carbon monoxide. Solketal visual appearance is transparent and it is odorless organic liquid used as fuel additive for diesel to improve its cold flow properties. Condensation of bio-glycerol with bio-acetone in presence of beta zeolite has been done for synthesizing solketal. It was observed that glycerol conversion and selectivity of solketal was largely effected by temperature, as it increases from 40 ºC to 60 ºC the conversion of glycerol rises from 80.04 % to 94.26 % and selectivity of solketal from 80.0 % to 94.21 % but further increase in temperature to 100 ºC glycerol conversion reduced to 93.06 % and solketal selectivity to 92.08 %. At the optimum conditions, the bio-glycerol conversion and solketal yield were about 94.26% and 94.21wt% respectively. This process offers an attractive route for converting bio-glycerol, the main by-product of biodiesel to solketal with bio-acetone; a value-added green product with potential industrial applications as a valuable green fuel additive or combustion promoter for gasoline/diesel engines.

Keywords: bio-acetone, bio-glycerol, acetylation, solketal

Procedia PDF Downloads 241
724 Study on Microbial Pretreatment for Enhancing Enzymatic Hydrolysis of Corncob

Authors: Kessara Seneesrisakul, Erdogan Gulari, Sumaeth Chavadej

Abstract:

The complex structure of lignocellulose leads to great difficulties in converting it to fermentable sugars for the ethanol production. The major hydrolysis impediments are the crystallinity of cellulose and the lignin content. To improve the efficiency of enzymatic hydrolysis, microbial pretreatment of corncob was investigated using two bacterial strains of Bacillus subtilis A 002 and Cellulomonas sp. TISTR 784 (expected to break open the crystalline part of cellulose) and lignin-degrading fungus, Phanerochaete sordida SK7 (expected to remove lignin from lignocellulose). The microbial pretreatment was carried out with each strain under its optimum conditions. The pretreated corncob samples were further hydrolyzed to produce reducing glucose with low amounts of commercial cellulase (25 U•g-1 corncob) from Aspergillus niger. The corncob samples were determined for composition change by X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), and scanning electron microscope (SEM). According to the results, the microbial pretreatment with fungus, P. sordida SK7 was the most effective for enhancing enzymatic hydrolysis, approximately, 40% improvement.

Keywords: corncob, enzymatic hydrolysis, glucose, microbial pretreatment

Procedia PDF Downloads 556
723 Synthesis of Oxygenated Fuel Additive from Bio-Glycerol

Authors: Farrukh Jamil, Ala'a H. Al-Muhtaseb, Lamya Al-Haj, Mohab A. Al-Hinai

Abstract:

Glycerol is considered as high boiling polar triol and immiscible with fossil fuel fractions due to which it is transformed into its respective ketals and acetals which help to improve the quality of diesel emitting less amount of aldehydes and carbon monoxide. Solketal visual appearance is transparent, and it is odorless organic liquid used as a fuel additive for diesel to improve its cold flow properties. Condensation of bio-glycerol with bio-acetone in presence of beta zeolite has been done for synthesizing solketal. It was observed that glycerol conversion and selectivity of solketal was largely effected by temperature, as it increases from 40 ºC to 60 ºC the conversion of glycerol rises from 80.04 % to 94.26 % and selectivity of solketal from 80.0 % to 94.21 % but further increase in temperature to 100 ºC glycerol conversion reduced to 93.06 % and solketal selectivity to 92.08 %. At the optimum conditions, the bio-glycerol conversion and solketal yield were about 94.26% and 94.21wt% respectively. This process offers an attractive route for converting bio-glycerol, the main by-product of biodiesel to solketal with bio-acetone; a value-added green product with potential industrial applications as a valuable green fuel additive or combustion promoter for gasoline/diesel engines.

Keywords: bio-glycerol, catalyst, green additive, biomass

Procedia PDF Downloads 219
722 An Algorithm to Find Fractional Edge Domination Number and Upper Fractional Edge Domination Number of an Intuitionistic Fuzzy Graph

Authors: Karunambigai Mevani Govindasamy, Sathishkumar Ayyappan

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In this paper, we formulate the algorithm to find out the dominating function parameters of Intuitionistic Fuzzy Graphs(IFG). The methodology we adopted here is converting any physical problem into an IFG, and that has been transformed into Intuitionistic Fuzzy Matrix. Using Linear Program Solver software (LiPS), we found the defined parameters for the given IFG. We obtained these parameters for a path and cycle IFG. This study can be extended to other varieties of IFG. In particular, we obtain the definition of edge dominating function, minimal edge dominating function, fractional edge domination number (γ_if^') and upper fractional edge domination number (Γ_if^') of an intuitionistic fuzzy graph. Also, we formulated an algorithm which is appropriate to work on LiPS to find fractional edge domination number and upper fractional edge domination number of an IFG.

Keywords: fractional edge domination number, intuitionistic fuzzy cycle, intuitionistic fuzzy graph, intuitionistic fuzzy path

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721 Mechanical Prosthesis Controlled by Brain-Computer Interface

Authors: Tianyu Cao, KIRA (Ruizhi Zhao)

Abstract:

The purpose of our research is to study the possibility of people with physical disabilities manipulating mechanical prostheses through brain-computer interface (BCI) technology. The brain-machine interface (BCI) of the neural prosthesis records signals from neurons and uses mathematical modeling to decode them, converting desired movements into body movements. In order to improve the patient's neural control, the prosthesis is given a natural feeling. It records data from sensitive areas from the body to the prosthetic limb and encodes signals in the form of electrical stimulation to the brain. In our research, the brain-computer interface (BCI) is a bridge connecting patients’ cognition and the real world, allowing information to interact with each other. The efficient work between the two is achieved through external devices. The flow of information is controlled by BCI’s ability to record neuronal signals and decode signals, which are converted into device control. In this way, we could encode information and then send it to the brain through electrical stimulation, which has significant medical application.

Keywords: biomedical engineering, brain-computer interface, prosthesis, neural control

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720 Oxidovanadium(IV) and Dioxidovanadium(V) Complexes: Efficient Catalyst for Peroxidase Mimetic Activity and Oxidation

Authors: Mannar R. Maurya, Bithika Sarkar, Fernando Avecilla

Abstract:

Peroxidase activity is possibly successfully used for different industrial processes in medicine, chemical industry, food processing and agriculture. However, they bear some intrinsic drawback associated with denaturation by proteases, their special storage requisite and cost factor also. Now a day’s artificial enzyme mimics are becoming a research interest because of their significant applications over conventional organic enzymes for ease of their preparation, low price and good stability in activity and overcome the drawbacks of natural enzymes e.g serine proteases. At present, a large number of artificial enzymes have been synthesized by assimilating a catalytic center into a variety of schiff base complexes, ligand-anchoring, supramolecular complexes, hematin, porphyrin, nanoparticles to mimic natural enzymes. Although in recent years a several number of vanadium complexes have been reported by a continuing increase in interest in bioinorganic chemistry. To our best of knowledge, the investigation of artificial enzyme mimics of vanadium complexes is very less explored. Recently, our group has reported synthetic vanadium schiff base complexes capable of mimicking peroxidases. Herein, we have synthesized monoidovanadium(IV) and dioxidovanadium(V) complexes of pyrazoleone derivateis ( extensively studied on account of their broad range of pharmacological appication). All these complexes are characterized by various spectroscopic techniques like FT-IR, UV-Visible, NMR (1H, 13C and 51V), Elemental analysis, thermal studies and single crystal analysis. The peroxidase mimic activity has been studied towards oxidation of pyrogallol to purpurogallin with hydrogen peroxide at pH 7 followed by measuring kinetic parameters. The Michaelis-Menten behavior shows an excellent catalytic activity over its natural counterparts, e.g. V-HPO and HRP. The obtained kinetic parameters (Vmax, Kcat) were also compared with peroxidase and haloperoxidase enzymes making it a promising mimic of peroxidase catalyst. Also, the catalytic activity has been studied towards the oxidation of 1-phenylethanol in presence of H2O2 as an oxidant. Various parameters such as amount of catalyst and oxidant, reaction time, reaction temperature and solvent have been taken into consideration to get maximum oxidative products of 1-phenylethanol.

Keywords: oxovanadium(IV)/dioxidovanadium(V) complexes, NMR spectroscopy, Crystal structure, peroxidase mimic activity towards oxidation of pyrogallol, Oxidation of 1-phenylethanol

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719 On the Other Side of Shining Mercury: In Silico Prediction of Cold Stabilizing Mutations in Serine Endopeptidase from Bacillus lentus

Authors: Debamitra Chakravorty, Pratap K. Parida

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Cold-adapted proteases enhance wash performance in low-temperature laundry resulting in a reduction in energy consumption and wear of textiles and are also used in the dehairing process in leather industries. Unfortunately, the possible drawbacks of using cold-adapted proteases are their instability at higher temperatures. Therefore, proteases with broad temperature stability are required. Unfortunately, wild-type cold-adapted proteases exhibit instability at higher temperatures and thus have low shelf lives. Therefore, attempts to engineer cold-adapted proteases by protein engineering were made previously by directed evolution and random mutagenesis. The lacuna is the time, capital, and labour involved to obtain these variants are very demanding and challenging. Therefore, rational engineering for cold stability without compromising an enzyme's optimum pH and temperature for activity is the current requirement. In this work, mutations were rationally designed with the aid of high throughput computational methodology of network analysis, evolutionary conservation scores, and molecular dynamics simulations for Savinase from Bacillus lentus with the intention of rendering the mutants cold stable without affecting their temperature and pH optimum for activity. Further, an attempt was made to incorporate a mutation in the most stable mutant rationally obtained by this method to introduce oxidative stability in the mutant. Such enzymes are desired in detergents with bleaching agents. In silico analysis by performing 300 ns molecular dynamics simulations at 5 different temperatures revealed that these three mutants were found to be better in cold stability compared to the wild type Savinase from Bacillus lentus. Conclusively, this work shows that cold adaptation without losing optimum temperature and pH stability and additionally stability from oxidative damage can be rationally designed by in silico enzyme engineering. The key findings of this work were first, the in silico data of H5 (cold stable savinase) used as a control in this work, corroborated with its reported wet lab temperature stability data. Secondly, three cold stable mutants of Savinase from Bacillus lentus were rationally identified. Lastly, a mutation which will stabilize savinase against oxidative damage was additionally identified.

Keywords: cold stability, molecular dynamics simulations, protein engineering, rational design

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718 Fast and Efficient Algorithms for Evaluating Uniform and Nonuniform Lagrange and Newton Curves

Authors: Taweechai Nuntawisuttiwong, Natasha Dejdumrong

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Newton-Lagrange Interpolations are widely used in numerical analysis. However, it requires a quadratic computational time for their constructions. In computer aided geometric design (CAGD), there are some polynomial curves: Wang-Ball, DP and Dejdumrong curves, which have linear time complexity algorithms. Thus, the computational time for Newton-Lagrange Interpolations can be reduced by applying the algorithms of Wang-Ball, DP and Dejdumrong curves. In order to use Wang-Ball, DP and Dejdumrong algorithms, first, it is necessary to convert Newton-Lagrange polynomials into Wang-Ball, DP or Dejdumrong polynomials. In this work, the algorithms for converting from both uniform and non-uniform Newton-Lagrange polynomials into Wang-Ball, DP and Dejdumrong polynomials are investigated. Thus, the computational time for representing Newton-Lagrange polynomials can be reduced into linear complexity. In addition, the other utilizations of using CAGD curves to modify the Newton-Lagrange curves can be taken.

Keywords: Lagrange interpolation, linear complexity, monomial matrix, Newton interpolation

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717 Preparation and Visible Light Photoactivity of N-Doped ZnO/ZnS Photocatalysts

Authors: Nuray Güy, Mahmut Özacar

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Semiconductor nanoparticles such as TiO₂ and ZnO as photocatalysts are very efficient catalysts for wastewater treatment by the chemical utilization of light energy, which is capable of converting the toxic and nonbiodegradable organic compounds into carbon dioxide and mineral acids. ZnO semiconductor has a wide bandgap energy of 3.37 eV and a relatively large exciton binding Energy (60 meV), thus can absorb only UV light with the wavelength equal to or less than 385 nm. It exhibits low efficiency under visible light illumination due to its wide band gap energy. In order to improve photocatalytic activity of ZnO under visible light, band gap of ZnO may be narrowed by doping such as N, C, S nonmetal ions and coupled two separate semiconductors possessing different energy levels for their corresponding conduction and valence bands. ZnS has a wider band gap (Eg=3.7 eV) than ZnO and generates electron–hole pairs by photoexcitation rapidly. In the present work, N doped ZnO/ZnS nano photocatalysts with visible-light response were synthesized by microwave-hydrothermal method using thiourea as N source. The prepared photocatalysts were characterized by X-ray diffraction (XRD), field emission scanning electron microscopy (FESEM) and UV–visible (UV–vis). The photocatalytic activities samples and undoped ZnO have been studied for the degradation of dye, and have also been compared with together.

Keywords: photocatalyst, synthesis, visible light, ZnO/ZnS

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716 Anti Corruption Conventions in Nigeria: Legal and Administrative Challenges

Authors: Mohammed Albakariyu Kabir

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There is a trend in development discourse to understand and explain the level of corruption in Nigeria, its anti-corruption crusade and why it is failing, as well as its level of compliance with International standards of United Nations Convention against Corruption (UNCAC) & African Union Convention on Converting and Preventing Corruption) to which Nigeria is a signatory. This paper discusses the legal and Constitutional provisions relating to corrupt practices and safeguards in Nigeria, as well as the obstacles to the implementation of these Conventions.The paper highlights the challenges posed to the Anti-Corruption crusade by analysing the loopholes that exist both in administrative structure and in scope of the relevant laws. The paper argues that Nigerian Constitution did not make adequate provisions for the implementation of the conventions, hence a proposal which will ensure adequate provision for implementing the conventions to better the lives of Nigerians. The paper concludes that there is the need to build institutional parameters, adequate constitutional and structural safeguards, as well as to synergise strategies, collaborations and alliances to facilitate the timely domestication and implementation of the conventions.

Keywords: anti-corruption, corruption, convention, domestication, poverty, state parties

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715 Advanced Digital Manufacturing: Case Study

Authors: Abdelrahman Abdelazim

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Most industries are looking for technologies that are easy to use, efficient and fast to accomplish. To implement these, factories tend to use advanced systems that could alter complicity to simplicity and rudimentary to advancement. Cloud Manufacturing is a new movement that aims to mirror and integrate cloud computing into manufacturing. Amongst cloud manufacturing various advantages are decreasing the human involvements and increasing the dependency on automated machines, which in turns decreases human errors and increases efficiency. A reliable and extraordinary performance processes with minimum errors are highly desired factors of today’s manufacturers. At the glance it seems to be the best alternative, however, the implementation of a cloud system can be very challenging. This work investigates cloud manufacturing in details, it outlines its advantages and disadvantages by converting a local factory in Kuwait to a cloud-ready system. Initially the flow of the factory’s manufacturing process has been analyzed identifying the bottlenecks and illustrating how cloud manufacturing can eliminate them. Following this an automation process has been analyzed and implemented. A comparison between the process before and after the adaptation has been carried out showing the effects on the cost, the output and the efficiency of the process.

Keywords: cloud manufacturing, automation, Kuwait industrial sector, advanced digital manufacturing

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714 Quick Sequential Search Algorithm Used to Decode High-Frequency Matrices

Authors: Mohammed M. Siddeq, Mohammed H. Rasheed, Omar M. Salih, Marcos A. Rodrigues

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This research proposes a data encoding and decoding method based on the Matrix Minimization algorithm. This algorithm is applied to high-frequency coefficients for compression/encoding. The algorithm starts by converting every three coefficients to a single value; this is accomplished based on three different keys. The decoding/decompression uses a search method called QSS (Quick Sequential Search) Decoding Algorithm presented in this research based on the sequential search to recover the exact coefficients. In the next step, the decoded data are saved in an auxiliary array. The basic idea behind the auxiliary array is to save all possible decoded coefficients; this is because another algorithm, such as conventional sequential search, could retrieve encoded/compressed data independently from the proposed algorithm. The experimental results showed that our proposed decoding algorithm retrieves original data faster than conventional sequential search algorithms.

Keywords: matrix minimization algorithm, decoding sequential search algorithm, image compression, DCT, DWT

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713 Gene Cloning and Expression of Azoreductases from Azo-Degraders Lysinibacillus macrolides and Bacillus coagulans Isolated from Egyptian Industrial Wastewater

Authors: Omaima A. Sharaf, Wafaa M. Abd El-Rahim, Hassan Moawad, Michael J. Sadowsky

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Textile industry is one of the important industries in the worldwide. It is known that the eco-friendly industrial and agricultural activities are significant for socio-economic stability of all countries. The absence of appropriate industrial waste water treatments is essential barrier for sustainable development in food and agricultural sectors especially in developing country like Egypt. Thus, the development of enzymatic bioremediation technology for textile dye removal will enhance the collaboration between scientists who develop the technology and industry where this technology will be implemented towards the safe disposal of the textile dye wastes. Highly efficient microorganisms are of most importance in developing and using highly effective biological treatment processes. Bacterial degradation of azo dyes is generally initiated by an enzymatic step that involves cleavage of azo linkages, usually with the aid of an azoreductase as electron donor. Thus, expanding the spectrum of microorganisms with high enzymatic activities as azoreductases and discovering novel azo-dye degrading enzymes, with enhanced stability and superior catalytic properties, are necessary for many environmental and industrial applications. Consequently, the use of molecular tools has become increasingly integrated into the understanding of enzyme properties and characterization. Researchers have utilized a gene cloning and expression methods as a tool to produce recombinant protein for decolorizing dyes more efficiently. Thus, presumptive evidence for the presence of genes encoding azoreductases in the genomes of selected local, and most potent azo-degrading strains were obtained by using specific oligonucleotides primers. These potent strains have been isolated from textile industrial wastewater in Egypt and identified using 16S rRNA sequence analysis as 'Lysinibacillus macrolidesB8, Brevibacillus parabrevisB11, Bacillus coagulansB7, and B. cereusB5'. PCR products of two full length genes designated as (AZO1;621bp and AZO2;534bp) were detected. BLASTx results indicated that AZO1 gene was corresponding to predicted azoreductase from of Bacillus sp. ABP14, complete genome, multispecies azoreductase [Bacillus], It was submitted to the gene bank by an accession no., BankIt2085371 AZO1 MG923210 (621bp; 207 amino acids). AZO1 was generated from the DNA of our identified strains Lysinibacillus macrolidesB8. On the other hand, AZO2 gene was corresponding to a predicted azoreductase from Bacillus cereus strain S2-8. Gene bank accession no. was BankIt2085839 AZO2 MG932081 (534bp;178 amino acids) and it was amplified from our Bacillus coagulansB7. Both genes were successfully cloned into pCR2.1TOPO (Invitrogen) and in pET28b+ vectors, then they transformed into E. coli DH5α and BL21(DE3) cells for heterologous expression studies. Our recombinant azoreductases (AZO1&AZO2) exhibited potential enzyme activity and efficiently decolorized an azo dye (Direct violet). They exhibited pH stability between 6 and 8 with optimum temperature up to 60°C and 37 °C after induction by 1mM and 1.5mM IPTG, for both AZO1 &AZO2, respectively. These results suggested that further optimization and purification of these recombinant proteins by using different heterologous expression systems will give great potential for the sustainable utilization of these recombinant enzymes in several industrial applications especially in wastewater treatments.

Keywords: azoreductases, decolorization, enzyme activity, gene cloning and expression

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712 Simplifying the Migration of Architectures in Embedded Applications Introducing a Pattern Language to Support the Workforce

Authors: Farha Lakhani, Michael J. Pont

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There are two main architectures used to develop software for modern embedded systems: these can be labelled as “event-triggered” (ET) and “time-triggered” (TT). The research presented in this paper is concerned with the issues involved in migration between these two architectures. Although TT architectures are widely used in safety-critical applications they are less familiar to developers of mainstream embedded systems. The research presented in this paper began from the premise that–for a broad class of systems that have been implemented using an ET architecture–migration to a TT architecture would improve reliability. It may be tempting to assume that conversion between ET and TT designs will simply involve converting all event-handling software routines into periodic activities. However, the required changes to the software architecture are, in many cases rather more profound. The main contribution of the work presented in this paper is to identify ways in which the significant effort involved in migrating between existing ET architectures and “equivalent” (and effective) TT architectures could be reduced. The research described in this paper has taken an innovative step in this regard by introducing the use of ‘Design patterns’ for this purpose for the first time.

Keywords: embedded applications, software architectures, reliability, pattern

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711 Efficiency Enhancement of Photovoltaic Panels Using an Optimised Air Cooled Heat Sink

Authors: Wisam K. Hussam, Ali Alfeeli, Gergory J. Sheard

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Solar panels that use photovoltaic (PV) cells are popular for converting solar radiation into electricity. One of the major problems impacting the performance of PV panels is the overheating caused by excessive solar radiation and high ambient temperatures, which degrades the efficiency of the PV panels remarkably. To overcome this issue, an aluminum heat sink was used to dissipate unwanted heat from PV cells. The dimensions of the heat sink were determined considering the optimal fin spacing that fulfils hot climatic conditions. In this study, the effects of cooling on the efficiency and power output of a PV panel were studied experimentally. Two PV modules were used: one without and one with a heat sink. The experiments ran for 11 hours from 6:00 a.m. to 5:30 p.m. where temperature readings in the rear and front of both PV modules were recorded at an interval of 15 minutes using sensors and an Arduino microprocessor. Results are recorded for both panels simultaneously for analysis, temperate comparison, and for power and efficiency calculations. A maximum increase in the solar to electrical conversion efficiency of 35% and almost 55% in the power output were achieved with the use of a heat sink, while temperatures at the front and back of the panel were reduced by 9% and 11%, respectively.

Keywords: photovoltaic cell, natural convection, heat sink, efficiency

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710 Generalized Correlation Coefficient in Genome-Wide Association Analysis of Cognitive Ability in Twins

Authors: Afsaneh Mohammadnejad, Marianne Nygaard, Jan Baumbach, Shuxia Li, Weilong Li, Jesper Lund, Jacob v. B. Hjelmborg, Lene Christensen, Qihua Tan

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Cognitive impairment in the elderly is a key issue affecting the quality of life. Despite a strong genetic background in cognition, only a limited number of single nucleotide polymorphisms (SNPs) have been found. These explain a small proportion of the genetic component of cognitive function, thus leaving a large proportion unaccounted for. We hypothesize that one reason for this missing heritability is the misspecified modeling in data analysis concerning phenotype distribution as well as the relationship between SNP dosage and the phenotype of interest. In an attempt to overcome these issues, we introduced a model-free method based on the generalized correlation coefficient (GCC) in a genome-wide association study (GWAS) of cognitive function in twin samples and compared its performance with two popular linear regression models. The GCC-based GWAS identified two genome-wide significant (P-value < 5e-8) SNPs; rs2904650 near ZDHHC2 on chromosome 8 and rs111256489 near CD6 on chromosome 11. The kinship model also detected two genome-wide significant SNPs, rs112169253 on chromosome 4 and rs17417920 on chromosome 7, whereas no genome-wide significant SNPs were found by the linear mixed model (LME). Compared to the linear models, more meaningful biological pathways like GABA receptor activation, ion channel transport, neuroactive ligand-receptor interaction, and the renin-angiotensin system were found to be enriched by SNPs from GCC. The GCC model outperformed the linear regression models by identifying more genome-wide significant genetic variants and more meaningful biological pathways related to cognitive function. Moreover, GCC-based GWAS was robust in handling genetically related twin samples, which is an important feature in handling genetic confounding in association studies.

Keywords: cognition, generalized correlation coefficient, GWAS, twins

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709 The Composition and Activity of Germinated Broccoli Seeds and Their Extract

Authors: Boris Nemzer, Tania Reyes-Izquierdo, Zbigniew Pietrzkowski

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Glucosinolate is a family of glucosides that can be found in a family of brassica vegetables. Upon the damage of the plant, glucosinolate breakdown by an internal enzyme myrosinase (thioglucosidase; EC 3.2.3.1) into isothiocyanates, such as sulforaphane. Sulforaphane is formed by glucoraphanin cleaving the sugar off by myrosinase and rearranged. Sulforaphane nitrile is formed in the same reaction as sulforaphane with the active of epithiospecifier protein (ESP). Most common food processing procedure would break the plant and mix the glucoraphanin and myrosinase together, and the formed sulforaphane would be further degraded. The purpose of this study is to understand the glucoraphanin/sulforaphane and the myrosinase activity of broccoli seeds germinated at a different time and technological processing conditions that keep the activity of the enzyme to form sulforaphane. Broccoli seeds were germinated in the house. Myrosinase activities were tested as the glucose content using glucose assay kit and measured UV-Vis spectrophotometer. Glucosinolates were measured by HPLC/DAD. Sulforaphane was measured using HPLC-DAD and GC/MS. The 6 hr germinated sprouts have a myrosinase activity 32.2 mg glucose/g, which is comparable with 12 and 24 hour germinated seeds and higher than dry seeds. The glucoraphanin content in 6 hour germinated sprouts is 13935 µg/g which is comparable to 24 hour germinated seeds and lower than the dry seeds. GC/MS results show that the amount of sulforaphane is higher than the amount of sulforaphane nitrile in seeds, 6 hour and 24 hour germinated seeds. The ratio of sulforaphane and sulforaphane nitrile is high in 6 hour germinated seeds, which indicates the inactivated ESP in the reaction. After evaluating the results, the short time germinated seeds can be used as the source of glucoraphanin and myrosinase supply to form potential higher sulforaphane content. Broccoli contains glucosinolates, glucoraphanin (4-methylsulfinylbutyl glucosinolate), which is an important metabolite with health-promoting effects. In the pilot clinical study, we observed the effects of a glucosinolates/glucoraphanin-rich extract from short time germinated broccoli seeds on blood adenosine triphosphate (ATP), reactive oxygen species (ROS) and lactate levels. A single dose of 50 mg of broccoli sprouts extract increased blood levels of ATP up to 61% (p=0.0092) during the first 2 hours after the ingestion. Interestingly, this effect was not associated with an increase in blood ROS or lactate. When compared to the placebo group, levels of lactate were reduced by 10% (p=0.006). These results indicate that broccoli germinated seed extract may positively affect the generation of ATP in humans. Due to the preliminary nature of this work and promising results, larger clinical trials are justified.

Keywords: broccoli glucosinolates, glucoraphanin, germinated seeds, myrosinase, adenosine triphosphate

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708 Solid Waste Management Challenges and Possible Solution in Kabul City

Authors: Ghulam Haider Haidaree, Nsenda Lukumwena

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Most developing nations face energy production and supply problems. This is also the case of Afghanistan whose generating capacity does not meet its energy demand. This is due in part to high security and risk caused by war which deters foreign investments and insufficient internal revenue. To address the issue above, this paper would like to suggest an alternative and affordable way to deal with the energy problem. That is by converting Solid Waste to energy. As a result, this approach tackles the municipal solid waste issue (potential cause of several diseases), contributes to the improvement of the quality of life, local economy, and so on. While addressing the solid waste problem in general, this paper samples specifically one municipality which is District-12, one of the 22 districts of Kabul city. Using geographic information system (GIS) technology, District-12 is divided into nine different zones whose municipal solid waste is respectively collected, processed, and converted into electricity and distributed to the closest area. It is important to mention that GIS has been used to estimate the amount of electricity to be distributed and to optimally position the production plant.

Keywords: energy problem, estimation of electricity, GIS zones, solid waste management system

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707 The Anti-Angiogenic Effect of Tectorigenin in a Mouse Model of Retinopathy of Prematurity

Authors: KuiDong Kang, Hye Bin Yim, Su Ah Kim

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Purpose: Tectorigenin is an isoflavone derived from the rhizome of Belamacanda chinensis. In this study, oxygen-induced retinopathy was used to characterize the anti-angiogenic properties of tectorigenin in mice. Methods: ICR neonatal mice were exposed to 75% oxygen from postnatal day P7 until P12 and returned to room air (21% oxygen) for five days (P12 to P17). Mice were subjected to daily intraperitoneal injection of tectorigenin (1 mg/kg, 10 mg/kg) and vehicle from P12 to P17. Retro-orbital injection of FITC-dextran was performed and retinal flat mounts were viewed by fluorescence microscopy. The Central avascular area was quantified from the digital images in a masked fashion using image analysis software (NIH ImageJ). Neovascular tufts were quantified by using SWIFT_NV and neovascular lumens were quantified from a histologic section in a masked fashion. Immunohistochemistry and Western blot analysis were also performed to demonstrate the anti-angiogenic activity of this compound in vivo. Results: In the retina of tectorigenin injected mouse (10mg/kg), the central non-perfusion area was significantly decreased compared to the vehicle injected group (1.76±0.5 mm2 vs 2.85±0.6 mm2, P<0.05). In vehicle-injected group, 33.45 ± 5.51% of the total retinal area was avascular, whereas the retinas of pups treated with high-dose (10 mg/kg) tectorigenin showed avascular retinal areas of 21.25 ±4.34% (P<0.05). High dose of tectorigenin also significantly reduced the number of vascular lumens in the histologic section. Tectorigenin (10 mg/kg) significantly reduced the expression of vascular endothelial growth factor (VEGF), matrix metalloproteinase-2 (MMP-2), MMP-9, and angiotensin II compared to the vehicle injected group. Tectorigenin did not affect CD31 abundance at any tested dose. Conclusions: Our results show that tectorigenin possesses powerful anti-angiogenic properties and can attenuate new vessel formation in the retina after systemic administration. These results imply that this compound can be considered as a candidate substance for therapeutic inhibition of retinal angiogenesis.

Keywords: tectorigenin, anti-angiogenic, retinopathy, Belamacanda chinensis

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706 Experimental Study of Solar Drying of Verbena in Three Types of Solar Dryers

Authors: Llham Lhoume, Rachid Tadili, Nora Arbaoui

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One of the most crucial ways to combat food insecurity is to minimize crop losses, food drying is one of the most organic, effective, low-cost and energy-efficient food preservation methods. In this regard, we undertake in this study an experimental evaluation and analysis of the thermal performance of different natural convection drying systems: a solar greenhouse dryer, an indirect solar dryer with a single compartment and a solar dryer with two compartments. These systems have been implemented at the Solar Energy and Environment Laboratory of Mohammed V University (Morocco). The objective of this work is to study the feasibility of converting a solar greenhouse into a solar dryer for use during the summer. On the other hand, to study the thermal performances of this greenhouse dryer by comparing it with other solar dryers. The experimental study showed that the drying of verbena leaves took 6 hours in the indirect dryer 1, 3 hours in the indirect dryer, 2 and 4 hours in the greenhouse dryer, but the amortization period of the solar greenhouse dryer is lower than the other two solar dryers. The results of this study provide key information on the implementation and performance of these systems for drying a food of great global interest.

Keywords: solar energy, drying, agriculture, biotechnologie

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705 New Recombinant Netrin-a Protein of Lucilia Sericata Larvae by Bac to Bac Expression Vector System in Sf9 Insect Cell

Authors: Hamzeh Alipour, Masoumeh Bagheri, Abbasali Raz, Javad Dadgar Pakdel, Kourosh Azizi, Aboozar Soltani, Mohammad Djaefar Moemenbellah-Fard

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Background: Maggot debridement therapy is an appropriate, effective, and controlled method using sterilized larvae of Luciliasericata (L.sericata) to treat wounds. Netrin-A is an enzyme in the Laminins family which secreted from salivary gland of L.sericata with a central role in neural regeneration and angiogenesis. This study aimed to production of new recombinant Netrin-A protein of Luciliasericata larvae by baculovirus expression vector system (BEVS) in SF9. Material and methods: In the first step, gene structure was subjected to the in silico studies, which were include determination of Antibacterial activity, Prion formation risk, homology modeling, Molecular docking analysis, and Optimization of recombinant protein. In the second step, the Netrin-A gene was cloned and amplified in pTG19 vector. After digestion with BamH1 and EcoR1 restriction enzymes, it was cloned in pFastBac HTA vector. It was then transformed into DH10Bac competent cells, and the recombinant Bacmid was subsequently transfected into insect Sf9 cells. The expressed recombinant Netrin-A was thus purified in the Ni-NTA agarose. This protein evaluation was done using SDS-PAGE and western blot, respectively. Finally, its concentration was calculated with the Bradford assay method. Results: The Bacmid vector structure with Netrin-A was successfully constructed and then expressed as Netrin-A protein in the Sf9 cell lane. The molecular weight of this protein was 52 kDa with 404 amino acids. In the in silico studies, fortunately, we predicted that recombinant LSNetrin-A have Antibacterial activity and without any prion formation risk.This molecule hasa high binding affinity to the Neogenin and a lower affinity to the DCC-specific receptors. Signal peptide located between amino acids 24 and 25. The concentration of Netrin-A recombinant protein was calculated to be 48.8 μg/ml. it was confirmed that the characterized gene in our previous study codes L. sericata Netrin-A enzyme. Conclusions: Successful generation of the recombinant Netrin-A, a secreted protein in L.sericata salivary glands, and because Luciliasericata larvae are used in larval therapy. Therefore, the findings of the present study could be useful to researchers in future studies on wound healing.

Keywords: blowfly, BEVS, gene, immature insect, recombinant protein, Sf9

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