Search results for: designed-laboratory activity

Authors: M. F. R. Zuthi, H. H. Ngo, W. S. Guo, S. S. Chen, N. C. Nguyen, L. J. Deng, T. D. C Tran

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Sustaining a desired rate of oxygen transfer for microbial activity is a matter of major concern for Biological Wastewater Treatment (MBR). The study reported in the paper was aimed at assessing the effects of microbial products on the Specific Oxygen Uptake Rate (SOUR) in a Conventional Membrane Bioreactor (CMBR) and that in a Sponge Submerged MBR (SSMBR). The production and progressive accumulation of Soluble Microbial Products (SMP) and Bound-Extracellular Polymeric Substances (BEPS) were found affecting the SOUR of the microorganisms which varied at different stages of operation of the MBR systems depending on the variable concentrations of the SMP/bEPS. The effect of bEPS on the SOUR was stronger in the SSMBR compared to that of the SMP, while relative high concentrations of SMP had adverse effects on the SOUR of the CMBR system. Of the different mathematical correlations analyzed in the study, logarithmic mathematical correlations could be established between SOUR and bEPS in SSMBR, and similar correlations could also be found between SOUR and SMP concentrations in the CMBR.

Keywords: microbial products, microbial activity, specific oxygen uptake rate, membrane bioreactor

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Authors: Pauline Nyssen, Ange Mouithys-Mickalad, Maryse Hoebeke

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Inflammation is a complex physiological phenomenon involving chemical and enzymatic mechanisms. Polymorphonuclear neutrophil leukocytes (PMNs) play an important role by producing reactive oxygen species (ROS) and releasing myeloperoxidase (MPO), a pro-oxidant enzyme. Released both in the phagolysosome and the extracellular medium, MPO produces during its peroxidase and halogenation cycles oxidant species, including hypochlorous acid, involved in the destruction of pathogen agents, like bacteria or viruses. Inflammatory pathologies, like rheumatoid arthritis, atherosclerosis induce an excessive stimulation of the PMNs and, therefore, an uncontrolled release of ROS and MPO in the extracellular medium, causing severe damages to the surrounding tissues and biomolecules such as proteins, lipids, and DNA. The treatment of chronic inflammatory pathologies remains a challenge. For many years, MPO has been used as a target for the development of effective treatments. Numerous studies have been focused on the design of new drugs presenting more efficient MPO inhibitory properties. However, some designed inhibitors can be toxic. An alternative consists of assessing the potential inhibitory action of clinically-known molecules, having antioxidant activity. Propofol, 2,6-diisopropyl phenol, which is used as an intravenous anesthetic agent, meets these requirements. Besides its anesthetic action employed to induce a sedative state during surgery or in intensive care units, propofol and its injectable form Diprivan indeed present antioxidant properties and act as ROS and free radical scavengers. A study has also evidenced the ability of propofol to inhibit the formation of the neutrophil extracellular traps fibers, which are important to trap pathogen microorganisms during the inflammation process. The aim of this study was to investigate the potential inhibitory action mechanism of propofol and Diprivan on MPO activity. To go into the anti-inflammatory action of propofol in-depth, two of its oxidative derivatives, 2,6-diisopropyl-1,4-p-benzoquinone (PPFQ) and 3,5,3’,5’-tetra isopropyl-(4,4’)-diphenoquinone (PPFDQ), were studied regarding their inhibitory action. Specific immunological extraction followed by enzyme detection (SIEFED) and molecular modeling have evidenced the low anti-catalytic action of propofol. Stopped-flow absorption spectroscopy and direct MPO activity analysis have proved that propofol acts as a reversible MPO inhibitor by interacting as a reductive substrate in the peroxidase cycle and promoting the accumulation of redox compound II. Overall, Diprivan exhibited a weaker inhibitory action than the active molecule propofol. In contrast, PPFQ seemed to bind and obstruct the enzyme active site, preventing the trigger of the MPO oxidant cycles. PPFQ induced a better chlorination cycle inhibition at basic and neutral pH in comparison to propofol. PPFDQ did not show any MPO inhibition activity. The three interest molecules have also demonstrated their inhibition ability on an important step of the inflammation pathway, the PMNs superoxide anions production, thanks to EPR spectroscopy and chemiluminescence. In conclusion, propofol presents an interesting immunomodulatory activity by acting as a reductive substrate in the peroxidase cycle of MPO, slowing down its activity, whereas PPFQ acts more as an anti-catalytic substrate. Although PPFDQ has no impact on MPO, it can act on the inflammation process by inhibiting the superoxide anions production by PMNs.

Keywords: Diprivan, inhibitor, myeloperoxidase, propofol, spectroscopy

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Authors: Sarekha Woranuch, Rangrong Yoksan

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Chitosan is a derivative of chitin, which is a second most naturally abundant polysaccharide found in crab shells, shrimp shells, and squid pens. The applications of chitosan in pharmaceutical, cosmetics, food and packaging industries have been reported owing to its general recognition as safe, excellent biodegradability and biocompatibility, as well as ability to form films, membranes, gels, beads, fibers and particles. Nevertheless, chitosan is an amino polysaccharide consisting of strong inter- and intramolecular hydrogen bonds which limit its solubility in neutral pH water resulting in restricted utilization. Chemical modification is an alternative way to impede hydrogen bond formation. The objective of the present research is to improve water solubility and antioxidant activity of chitosan by grafting with ferulic acid. Ferulic acid was grafted onto chitosan at the C-2 position via a carbodiimide-mediated coupling reaction. Different mole ratios of chitosan to ferulic acid (i.e. 1.0:0.0, 1.0:0.5, 1.0:1.0, 1.0:1.5, 1.0:2.0, and 1.0:2.5) and various reaction temperatures (i.e. 40, 60, and 80 °C) were used. The reaction was performed at different times (i.e. 1.5, 3.0, 4.5, and 6.0 h). The obtained ferulic acid-grafted chitosan was characterized by FTIR and 1H NMR technique. The influences of ferulic acid on crystallinity, solubility and radical scavenging activity of chitosan were also investigated. Ferulic acid grafted chitosan was successfully synthesized as confirmed from (i) the appearance of FTIR absorption band at 1517 cm-1 belonging to C=C aromatic ring of ferulic acid and the increased C–H stretching band intensity and (ii) the appearance of proton signals at δ = 6.31-7.67 ppm ascribing to methine protons of ferulic acid. The condition in which the reaction temperature of 60°C, reaction time of 3 h and the mole ratio of chitosan to ferulic acid of 1:1 gave the highest ferulic acid substitution degree, i.e. 0.37. The resulting ferulic acid grafted chitosan was soluble in water (1.3 mg/mL) due to its reduced crystallinity as compared with chitosan and also exhibited 90% greater radical scavenging activity than chitosan. The result suggested the utilization of ferulic acid grafted chitosan as an antioxidant material.

Keywords: antioxidant property, chitosan, ferulic acid, grafting

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Authors: Eden G. Mariquit, Winarto Kurniawan, Masahiro Miyauchi, Hirofumi Hinode

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This research studied the effect of adding surfactant to the titanium dioxide (TiO2) sol-gel solution that was used to immobilize TiO2 on glass substrates by dip coating technique using TiO2 sol-gel solution mixed with different types of surfactants. After dipping into the TiO2 sol, the films were calcined and produced pure anatase crystal phase. The thickness of the thin film was varied by repeating the dip and calcine cycle. The prepared films were characterized using FE-SEM, TG-DTA, and XRD, and its photocatalytic performances were tested on degradation of an organic dye, methylene blue. Aside from its phocatalytic performance, the photo-induced hydrophilicity of thin TiO2 films surface was also studied. Characterization results showed that the addition of surfactant gave rise to characteristic patterns on the surface of the TiO2 thin film which also affects the photocatalytic activity. The addition of CTAB to the TiO2 dipping solution had a negative effect because the calcination temperature was not high enough to burn all the surfactants off. As for the surface wettability, the addition of surfactant also affected the induced surface hydrophilicity of the TiO2 films when irradiated under UV light.

Keywords: photocatalysis, surface hydrophilicity, TiO2 thin films, surfactant

Procedia PDF Downloads 418

Authors: Aka S. Koffi, N’Goran Yao, Philippe Bastide, Denis Bruneau, Diby Kadjo

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Cocoa beans (Theobroma cocoa L.) are the main components for chocolate manufacturing. The beans must be correctly fermented at first. Traditional process to perform the first fermentation (lactic fermentation) often consists in confining cacao beans using banana leaves or a fermentation basket, both of them leading to a poor product thermal insulation and to an inability to mix the product. Box fermenter reduces this loss by using a wood with large thickness (e>3cm), but mixing to homogenize the product is still hard to perform. Automatic fermenters are not rentable for most of producers. Heat (T>45°C) and acidity produced during the fermentation by microbiology activity of yeasts and bacteria are enabling the emergence of potential flavor and taste of future chocolate. In this study, a cylindro-rotative fermenter (FCR-V1) has been built and coconut fibers were used in its structure to confine heat. An axis of rotation (360°) has been integrated to facilitate the turning and homogenization of beans in the fermenter. This axis permits to put fermenter in a vertical position during the anaerobic alcoholic phase of fermentation, and horizontally during acetic phase to take advantage of the mid height filling. For circulation of air flow during turning in acetic phase, two woven rattan with grid have been made, one for the top and second for the bottom of the fermenter. In order to reduce air flow during acetic phase, two airtight covers are put on each grid cover. The efficiency of the turning by this kind of rotation, coupled with homogenization of the temperature, caused by the horizontal position in the acetic phase of the fermenter, contribute to having a good proportion of well-fermented beans (83.23%). In addition, beans’pH values ranged between 4.5 and 5.5. These values are ideal for enzymatic activity in the production of the aromatic compounds inside beans. The regularity of mass loss during all fermentation makes it possible to predict the drying surface corresponding to the amount being fermented.

Keywords: cocoa fermentation, fermenter, microbial activity, temperature, turning

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Authors: Zhaojun Zheng, Yuanfa Liu, Jiaxin Li, Jinwei Li, Yong-jiang Xu, Chen Cao

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Black bean is an excellent protein source for preparing hydrolysates, which attract much attention due to their biological activity. The objective of this study was to characterize the physicochemical and antioxidant properties of black bean protein, hydrolyzed by ficin, bromelain or alcalase until 300 min of hydrolysis. Results showed that bromelain and alcalase hydrolysates possessed a higher degree of hydrolysis (DH) than that of ficin, thereby presenting different ultraviolet absorption, fluorescence intensity, and circular dichroism. Moreover, all hydrolysates possessed the capacity to scavenge DPPH radical with the lowest IC₅₀ of 21.11 µg/mL, as well as to chelate ferrous ion (Fe²⁺) with the IC₅₀ values ranging from 6.82 to 30.68 µg/mL. Intriguingly, the oxidation of linoleic acid, sunflower oil, and sunflower oil-in-water emulsion was remarkedly retarded by the three selected protein hydrolysates, especially by bromelain-treated protein hydrolysate, which might attribute to their high hydrophobicity and emulsifying properties. These findings can provide strong support for black bean protein hydrolysates to be employed in food products acting as natural antioxidant alternatives.

Keywords: antioxidant activity, black bean protein hydrolysate, emulsion physicochemical properties, sunflower oil

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Authors: Ramón Aparicio-García, Gustavo Juárez Gracia, Jesús Álvarez Cedillo

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A research line of the computer science that involve the study of the Human-Computer Interaction (HCI), which search to recognize and interpret the user intent by the storage and the subsequent analysis of the electrical signals of the brain, for using them in the control of electronic devices. On the other hand, the affective computing research applies the human emotions in the HCI process helping to reduce the user frustration. This paper shows the results obtained during the hardware and software development of a Brain Computer Interface (BCI) capable of recognizing the human emotions through the association of the brain electrical activity patterns. The hardware involves the sensing stage and analogical-digital conversion. The interface software involves algorithms for pre-processing of the signal in time and frequency analysis and the classification of patterns associated with the electrical brain activity. The methods used for the analysis and classification of the signal have been tested separately, by using a database that is accessible to the public, besides to a comparison among classifiers in order to know the best performing.

Keywords: affective computing, interface, brain, intelligent interaction

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Authors: Rosina U. Ndukwe, Diane Crone, Nick Cavill

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Active transport (i.e. walking to school, cycle to work schemes etc.) is an effective approach with multiple social and environmental benefits for transforming urban environments into active urban environments. Although walking and cycling often remain on the margins of urban planning and infrastructure, there are new approaches emerging, along with policy intervention relevant for the creation of sustainable urban active environments conductive to active travel, increasing physical activity levels of involved communities and supporting social inclusion through more active participation. SPAcE - Supporting Policy and Action for Active Environments is a 3 year Erasmus+ project that aims to integrate active transport programmes into public policy across the EU. SPAcE focuses on cities/towns with recorded low physical activity levels to support the development of active environments in 5 sites: Latvia [Tukums], Italy [Palermo], Romania [Brasov], Spain [Castilla-La Mancha] and Greece [Trikala]. The first part of the project involved a review of good practice including case studies from across the EU and project partner countries. This has resulted in the first output from the project, an evidence of good practice summary with case study examples. In the second part of the project, working groups across the 5 sites have carried out co-production to develop Urban Active Environments (UActivE) Action Plans aimed at influencing policy and practice for increasing physical activity primarily through the use of cycling and walking. Action plans are based on international evidence and guidance for healthy urban planning. Remaining project partners include Universities (Gloucestershire, Oxford, Zurich, Thessaly) and Fit for Life programme (National physical activity promotion program, Finland) who provide support and advice incorporating current evidence, healthy urban planning and mentoring. Cooperation and co-production with public health professionals, local government officers, education authorities and transport agencies has been a key approach of the project. The third stage of the project has involved training partners in the WHO HEAT tool to support the implementation of the Action Plans. Project results show how multi-agency, transnational collaboration can produce real-life Action Plans in five EU countries, based on published evidence, real-life experience, consultation and collaborative working with other organisations across the EU. Learning from the processes adopted within this project will demonstrate how public health, local government and transport agencies across the EU, can work together to create healthy environments that have the aim of facilitating active behaviour, even in times of constrained public budgets. The SPAcE project has captured both the challenges and solutions for increasing population physical activity levels, health and wellness in urban spaces and translating evidence into policy and practice ensuring innovation at policy level. Funding acknowledgment: SPAcE (www.activeenvironments.eu) is co-funded by the Sport action of the ERASMUS+ programme.

Keywords: action plans, active transport, SPAcE, UActivE urban active environments, walking and cycling

Procedia PDF Downloads 264

Authors: Sibghat Mansoor Rana, Muhammad Islam, Hamid Saeed, Hummera Rafique, Muhammad Majid, Muhammad Tahir Aqeel, Fariha Imtiaz, Zaman Ashraf

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The 1,3,4-oxadiazole derivatives Ox-6a-f have been synthesized by incorporating flur- biprofen moiety with the aim to explore the potential of target molecules to decrease the oxidative stress. The title compounds Ox-6a-f were prepared by simple reactions in which a flurbiprofen –COOH group was esterified with methanol in an acid-catalyzed medium, which was then reacted with hydrazine to afford the corresponding hydrazide. The acid hydrazide was then cyclized into 1,3,4-oxadiazole-2-thiol by reacting with CS2 in the presence of KOH. The title compounds Ox-6a-f were synthesized by the reaction of an –SH group with various alkyl/aryl chlorides, which involves an S-alkylation reaction. The structures of the synthesized Ox-6a-f derivatives were ascer- tained by spectroscopic data. The in silico molecular docking was performed against target proteins cyclooxygenase-2 COX-2 (PDBID 5KIR) and cyclooxygenase-1 COX-1 (PDBID 6Y3C) to determine the binding affinity of the synthesized compounds with these structures. It has been inferred that most of the synthesized compounds bind well with an active binding site of 5KIR compared to 6Y3C, and especially compound Ox-6f showed excellent binding affinity (7.70 kcal/mol) among all synthesized compounds Ox-6a-f. The molecular dynamic (MD) simulation has also been performed to check the stability of docking complexes of ligands with COX-2 by determining their root mean square deviation and root mean square fluctuation. Little fluctuation was observed in case of Ox-6f, which forms the most stable complex with COX-2. The comprehensive antioxidant potential of the synthesized compounds has been evaluated by determining their free radical scavenging activity, including DPPH, OH, nitric oxide (NO), and iron chelation assay. The derivative Ox-6f showed promising results with 80.23% radical scavenging potential at a dose of 100 μg/mL while ascorbic acid exhibited 87.72% inhibition at the same dose. The anti-inflammatory activity of the final products has also been performed, and inflammatory markers were assayed, such as a thiobarbituric acid-reducing substance, nitric oxide, interleukin-6 (IL-6), and COX-2. The derivatives Ox-6d and Ox-6f displayed higher anti-inflammatory activity, exhibiting 70.56% and 74.16% activity, respectively. The results were compared with standard ibuprofen, which showed 84.31% activity at the same dose, 200 μg/mL. The anti-inflammatory potential has been performed by following the carrageen-induced hind paw edema model, and results showed that derivative Ox-6f exhibited 79.83% reduction in edema volume compared to standard ibuprofen, which reduced 84.31% edema volume. As dry lab and wet lab results confirm each other, it has been deduced that derivative Ox-6f may serve as the lead structure to design potent compounds to address oxidative stress.

Keywords: synthetic chemistry, pharmaceutical chemistry, oxadiazole derivatives, anti-inflammatory, anti-cancer compounds

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Authors: R. Majidzadeh Heravi, M. Sankian, H. Kermanshahi, M. R. Nassiri, A. Heravi Moussavi, S. A. Lari, A. R. Varasteh

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The use of probiotics engineered to express specific enzymes has been the subject of considerable attention in poultry industry because of increased nutrient availability and reduced cost of enzyme supplementation. Phytase enzyme is commonly added to poultry feed to improve digestibility and availability of phosphorus from plant sources. To construct a probiotic with potential of phytate degradation, phytase gene (appA) from E. coli was cloned and transformed into two probiotic bacteria Lactobacillus salivarius and Lactococcus lactis. L. salivarous showed plasmid instability, unable to express the gene. The expression of appA gene in L. lactis was analyzed by detecting specific RNA and zymography assay. Phytase enzyme was isolated from cellular extracts of recombinant L. lactis, showing a 46 kDa band upon the SDS-PAGE analysis. Zymogram also confirmed the phytase activity of the 46 kDa band corresponding to the enzyme. An enzyme activity of 4.9U/ml was obtained in cell extracts of L. lactis. The growth of native and recombinant L. lactis was similar in the presence of two concentrations of ox bile.

Keywords: Lactobacillus salivarus, Lactococcuslactis, recombinant, phytase, poultry

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Authors: Fatima Ammari, Meriem Chenouf

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Synthesis of gold nano particles has attracted much attention since the pioneering discovery of the high catalytic activity of supported gold nano particles in the reaction of CO oxidation at low temperature. In this research field, we used Na-montmorillonite for gold nanoparticles stabilization; different loading percentage 1, 2 and 5%. The gold nano particles were obtained using chemical reduction method using NaBH4 as reductant agent. The obtained gold nano particles Au-mont stabilized in Na-montmorillonite were used as catalysts for reduction of 4-nitrophenol to aminophenol with sodium borohydride at room temperature. The UV-Vis results confirm directly the gold nano particles formation. The XRD and N2 adsorption results showed the formation of gold nano particles in the pores of montmorillonite with an average size of 5 nm obtained on samples with 2%Au-mont. The gold particles size increased with the increase of gold loading percentage. The reduction reaction of 4-nitrophenol into 4-aminophenol with NaBH4 catalyzed by Au-Na-montmorillonite catalyst exhibits remarkably a high activity; the reaction was completed within 9 min for 1Au-mont and within 3 min for 2Au-mont.

Keywords: chemical reduction, gold, montmorillonite, nano particles, 4-nitrophenol

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Authors: Wagner Pereira, Alphonse Kelecom

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Mining can impact the environment, and the major impact of some mining activities is the radiological impact. In human populations, such impact is well studied and regulated. For biota, this assessment always had as focus the protection of human food chain. The protection of biota itself is a new approach, still developing. In order to contribute to this new approach, fish collecting was carried out in areas of naturally occurring radioactive materials (NORM), where a uranium mine is in decommissioning phase. The activity concentrations were analyzed, in Bq/kg wet weight, for Uranium (Unat), Th-232 and Ra-226 in the lambari fish Astyanax bimaculatus L. (omnivorous fish) and in the traíra fish Hoplias malabaricus Bloch, 1794 (carnivorous fish). Seven composite samples (that is: a sufficient number of individuals to reach at least 2 kg of fresh weight) were collected every six months between 2013 and 2015. The mean activity concentrations (AC) for uranium ranged from 1.12 (lambari) to 0.60 (lungfish). For Th, variations ranged from 0.30 to 0.05 (lambari and traíra, respectively). Finally, the Ra-226 means ranged between 0.08 and 0.03. No temporal trends of accumulation could be identified. Systematically, the AC values of radionuclides were higher in omnivorous fish when compared to the carnivore ones.

Keywords: biota dose, NORM, fish, environmental protection

Procedia PDF Downloads 258

Authors: A. H. Taghdisi, M. Mirmohammadi, S. Kamali

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Evening primrose (Oenothera biennis L.) is a biennial and herbaceous and one of the most important species of medicinal plants in the world. due to the production of unsaturated fatty acids such as linoleic acid, alpha-linolenic acid, etc. in its seeds and roots, and compounds such as kaempferol in its leaves, Evening primrose has important medicinal efficiency such as reducing premenstrual problems, acceleration of wound healing, inhibiting platelet aggregation, sedation of cardiovascular diseases, and treatment of viral infections. The sap of the plant is used to treat warts, and the plant itself is used as a charm against mental and spiritual diseases and poisonous animals. Its leaves have significant antibacterial activity against yellow staphylococci. It is also used in the treatment of poisoning, especially the toxication caused by the consumption of alcoholic beverages, in the treatment of arteriosclerosis and diseases caused by liver cell insufficiency. Low germination and production speed are the problems of evening primrose growth and propagation. In the present study, extracts were obtained from four components (flowers, stems, seeds, leaves) of the evening primrose plant using the Soxhlet apparatus. To measure the antibacterial properties against MDR bacteria, microbial methods, including dilution, cultivation on a plate containing nutrient agar culture medium, and disc diffusion in agar, were performed using Staphylococcus aureus and Escherichia coli bacteria on all four extracts. The maximum antibacterial activity related to the dilution method was obtained in all extracts. In the plate culture method, antibacterial activity was obtained for all extracts in the nutrient agar medium. The maximum diameter of the non-growth halo was obtained in the disc diffusion method in agar in the leaf extract. The statistical analysis of the microbial part was done by one-way ANOVA test (SPSS). By comparing the amount of omega-3 in extracts of Iranian and foreign oils available in the market and the extracts extracted from evening primrose plant samples with gas chromatography, it is shown that the stem extract had the most omega-3 (oleic acid) and compared to the extract of the mentioned oils, it had the highest amount of omega-3 overall. Also, the amount of omega-3 in the extract of Iranian oils was much higher than in the extract of foreign oils. It should be noted that the extract of foreign oils had a more complete composition of omega-3 than the extract of Iranian oils.

Keywords: antibacterial activity, MDR bacteria, evening primrose, omega-3

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Authors: R. P. Hewawasam, A. F. Dulhunty

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The ryanodine receptor (RyR) is an intracellular ion channel that releases Ca2+ from the sarcoplasmic reticulum and is essential for the excitation-contraction coupling and contraction in striated muscle. Human muscle specific glutathione transferase M2-2 (GSTM2-2) is a highly specific inhibitor of cardiac ryanodine receptor (RyR2) activity. Single channel-lipid bilayer studies and Ca2+ release assays performed using the C-terminal half of the GSTM2-2 and its mutants F157A and Y160A confirmed the ability of the C terminal domain of GSTM2-2 to specifically inhibit the cardiac ryanodine receptor activity. Objective of the present study is to determine the effect of C terminal domain of GSTM2-2 (GSTM2-2C) and the mutants, F157A and Y160A on the Ca2+ transients of neonatal ventricular cardiomyocytes. Primary cardiomyocytes were cultured from neonatal rats. They were treated with GSTM2-2C and the two mutants F157A and Y160A at 15µM and incubated for 2 hours. Then the cells were led with Fluo-4AM, fluorescent Ca2+ indicator, and the field stimulated (1 Hz, 3V and 2ms) cells were excited using the 488 nm argon laser. Contractility of the cells were measured and the Ca2+ transients in the stained cells were imaged using Leica SP5 confocal microscope. Peak amplitude of the Ca2+ transient, rise time and decay time from the peak were measured for each transient. In contrast to GSTM2C which significantly reduced the % shortening (42.8%) in the field stimulated cells, F157A and Y160A failed to reduce the % shortening.Analysis revealed that the average amplitude of the Ca2+ transient was significantly reduced (P<0.001) in cells treated with the wild type GSTM2-2C compared to that of untreated cells. Cells treated with the mutants F157A and Y160A didn’t change the Ca2+ transient significantly compared to the control. A significant increase in the rise time (P< 0.001) and a significant reduction in the decay time (P< 0.001) were observed in cardiomyocytes treated with GSTM2-2C compared to the control but not with F157A and Y160A. These results are consistent with the observation that GSTM2-2C reduced the Ca2+ release from the cardiac SR significantly whereas the mutants, F157A and Y160A didn’t show any effect compared to the control. GSTM2-2C has an isoform-specific effect on the cardiac ryanodine receptor activity and also it inhibits RyR2 channel activity only during diastole. Selective inhibition of RyR2 by GSTM2-2C has significant clinical potential in the treatment of cardiac arrhythmias and heart failure. Since GSTM2-2C-terminal construct has no GST enzyme activity, its introduction to the cardiomyocyte would not exert any unwanted side effects that may alter its enzymatic action. The present study further confirms that GSTM2-2C is capable of decreasing the Ca2+ release from the cardiac SR during diastole. These results raise the future possibility of using GSTM2-2C as a template for therapeutics that can depress RyR2 function when the channel is hyperactive in cardiac arrhythmias and heart failure.

Keywords: arrhythmia, cardiac muscle, cardiac ryanodine receptor, GSTM2-2

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Authors: Radia Imane Fertout

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In recent years, the reaction of dry reforming of methane (DRM) has attracted much attention due to its environmental and industrial importance. Various catalysts, including Ni-based catalysts, have been investigated for the DRM. Doping Ni/Al₂O₃ by lanthanum and alkaline earth element may strongly influence solid-state reaction and increases the stability of catalysts due to the lower density and high basicity of these oxides. The effect of SrO on the activity and stability of Ni/Al₂O₃-La₂O₃ in dry reforming of methane was investigated. These catalysts have been prepared with the impregnation method, calcined in air at 450 and 650°C, then characterized by BET surface area, X-ray diffraction (XRD), and scanning electron microscopy (SEM) techniques and tested in DRM. The results showed that the addition of strontium to Ni/Al2O₃-La₂O₃ decreased the specific surface area. XRD results revealed the presence of different phases of Al₂O₃, La(OH)₃, La₂O₂CO₃, and SrCO₃. The catalytic evaluation results showed that adding SrO increased the catalytic activity and stability, that explained by the strong basicity of strontium. SEM analysis after the reaction indicates the formation of carbon over the spent catalyst and that the addition of strontium stabilized the surface of the catalyst.

Keywords: dry reforming of methane, Ni/Al₂O₃-La₂O₃ catalyst, strontium, nickel

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Authors: Oluwaseun A. Oyetade, Roelof J. Kriek

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The development of effective catalysts for the oxygen evolution reaction (OER) is of great importance to combat energy-related concerns in the environment. Herein, we report a one-step solvothermal method employed for the fabrication of nickel selenide hybrids (NiSe-Ni₃Se₂) and a series of nickel selenide hybrid/multiwalled carbon nanotube composites (NiSe-Ni₃Se₂/MWCNT) as electrocatalysts for OER in alkaline media. The catalytic activities of these catalysts were investigated via several electrochemical characterization techniques, such as linear sweep voltammetry, chronoamperometric studies at constant potential, electrochemical surface area determination, and Tafel slope calculation, under alkaline conditions. Morphological observations demonstrated the agglomeration of non-uniform NiSe-Ni₃Se₂ microspheres around carbon nanotubes (CNTs), demonstrating the successful synthesis of NiSe-Ni₃Se₂/MWCNT nanocomposites. Among the tested electrocatalysts, the 20% NiSe-Ni₃Se₂/MWCNT nanocomposite demonstrated the highest activity, exhibiting an overpotential of 325 mV to achieve a current density of 10 mA.cm⁻² in 0.1 mol.dm⁻³ KOH solution. The NiSe-Ni₃Se₂/MWCNT nanocomposites showed improved activity toward OER compared to bare NiSe-Ni₃Se₂ hybrids and MWCNTs, exhibiting an overpotential of 528, 392 and 434 mV for 10%, 30% and 50% NiSe-Ni₃Se₂/MWCNT nanocomposites, respectively. These results compare favourably to the overpotential of noble catalysts, such as RuO₂ and IrO₂. Our results imply that the addition of MWCNTs increased the activity of NiSe-Ni₃Se₂ hybrids due to an increased number of catalytic sites, dispersion of NiSe-Ni₃Se₂ hybrid nanoparticles, and electronic conductivity of the nanocomposites. These nanocomposites also demonstrated better long-term stability compared to NiSe-Ni₃Se₂ hybrids and MWCNTs. Hence, NiSe-Ni₃Se₂/MWCNT nanocomposites possess the potential as effective electrocatalysts for OER in alkaline media.

Keywords: carbon nanotubes, electrocatalysts, nanocomposites, nickel selenide hybrids, oxygen evolution reaction

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Authors: Benedict Ofori, Kwabena Sarpong, Stephen Antwi

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Background: Medicinal plants are utilized all around the world and are becoming increasingly important economically. The WHO notes that ‘inappropriate use of traditional medicines or practices can have negative or dangerous effects and that future research is needed to ascertain the efficacy and safety of such practices and medicinal plants used by traditional medicine systems. The poor around the world have limited access to palliative care or pain relief. Pharmacologists have been focused on developing safe and effective anti-inflammatory drugs. Most of the issues related to their use have been linked to the fact that numerous traditional and herbal treatments are classified in different nations as meals or dietary supplements. As a result, there is no need for evidence of the quality, efficacy, or safety of these herbal formulations before they are marketed. The fact that access to drugs meant for pain relief is limited in low-income countries means advanced studies should be done on home drugs meant for inflammation to close the gap. Methods: The ethanolic extracts of the plant were screened for the presence of 10 phytochemicals. The Pierce BCA Protein Assay Kit was used for the determination of the protein concentration of the egg white. The rats were randomly selected and put in 6 groups. The egg white was sub-plantar injected into the right-hand paws of the rats to induce inflammation. The animals were treated with the three plant extracts obtained from the root bark, stem, and leaves of the plant. The control groups were treated with normal saline, while the standard groups were treated with standard drugs indomethacin and celecoxib. Plethysmometer was used to measure the change in paw volume of the animals over the course of the experiment. Results: The results of the phytochemical screening revealed the presence of reducing sugars and saponins. Alkaloids were present in only R.L.S (1:1:1), and phytosterols were found in R.L(1:1) and R.L.S (1:1:1). The estimated protein concentration was found to be 103.75 mg/ml. The control group had an observable increase in paw volume, which indicated that inflammation was induced during the 5 hours. The increase in paw volume for the control group peaked at the 1st hour and decreased gradually throughout the experiment, with minimal changes in the paw volumes. The 2nd and 3rd groups were treated with 20 mg/kg of indomethacin and celecoxib. The anti-inflammatory activities of indomethacin and celecoxib were calculated to be 21.4% and 4.28%, respectively. The remaining 3 groups were treated with 2 dose levels of 200mg/kg plant extracts. R.L.S, R.L, and S.R.L had anti-inflammatory activities of 22.3%, 8.2%, and 12.07%, respectively. Conclusions: Egg albumin-induced paw model in rats can be used to evaluate the anti-inflammatory activity of herbs that might have potential anti-inflammatory activity. Herbal medications have potential anti-inflammatory activities and can be used to manage various inflammatory conditions if their efficacy and side effects are well studied. The three extracts all possessed anti-inflammatory activity, with R.L.S having the highest anti-inflammatory activity.

Keywords: inflammation, capparis erythrocarpos, anti-inflammatory activity, herbal medicine, paw volume, egg albumin

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Authors: Paul Awolade, Parvesh Singh

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The global threat of pathogenic resistance to available therapeutic agents has become a menace to clinical practice, public health and man’s existence inconsequential. This has therefore led to an exigency in the development of new molecular scaffolds with profound activity profiles. In this vein, a versatile synthetic tool for accessing new molecules by incorporating two or more pharmacophores into a single entity with the unique ability to be recognized by multiple receptors hence leading to an improved bioactivity, known as molecular hybridization, has been explored with tremendous success. Accordingly, aware of the similarity in pharmacological activity spectrum of quinoline and 1,2,3-triazole pharmacophores such as; anti-Alzheimer, anticancer, anti-HIV, antimalarial and antimicrobial to mention but a few, the present study sets out to synthesize hybrids of quinoline and 1,2,3-triazole. The hybrids were accessed via click chemistry using copper catalysed azide-alkyne 1,3-dipolar cycloaddition reaction. All synthesized compounds were evaluated for their pharmaco-potential in an antimicrobial assay out of which the 3-OH derivative emerged as the most active with MIC value of 4 μg/mL against Cryptococcus neoformans; a value superior to standard Fluconazole and comparable to Amphotericin B. Structures of synthesized hybrids were elucidated using appropriate spectroscopic techniques (1H, 13C and 2D NMR, FT-IR and HRMS).

Keywords: bioisostere, click chemistry, molecular hybridization, quinoline, 1, 2, 3-triazole

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Authors: Tesleem Adewale Ibrahim

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Introduction: The prevalence of methicillin-resistant Staphylococcus aureus (MRSA) has been slowly rising in Nigeria for the past few decades. Therefore, novel classes of antibiotics are indispensable to combat the increased incidence of newly emerging multidrug-resistant bacteria like MRSA. Plants have been commonly used in popular medicine of most cultures for the treatment of disease. The in vitro antibacterial activity of some Nigerian common medicinal plants used in traditional medicine has been reported. The aim of this study was to investigate the antibacterial and anti-biofilm of these native plants (Entada abysinnica (leaves), Croton macrostachyus (leaves), Bridelia speciosa (seeds, bark), and Aframomum melegueta (leaves, seeds, and stem) collected in Southwestern Nigeria against a panel of seven biofilm-forming MRSA. Methods: Minimum inhibitory concentrations (MIC) and minimum bactericidal concentrations (MBC) of the plant extracts against MRSA were determined by the broth dilution method, and the anti-biofilm assay of the most potent plant extract was performed. Result: The results revealed that, of the four plants, water extracts of leaves of Entada abysinnica, leaves of Croton macrostachyus, seeds and bark Bridelia speciosa, and seeds of Aframomum melegueta exhibited significant antibacterial activity. Based on the MIC/MBC ratio, the extracts of these plants were determined to be bacteriostatic in nature. Anti-biofilm assay showed that the extract of seeds of Aframomum melegueta and leaves of Croton macrostachyus fairly inhibited the growth of MRSA in the preformed biofilm matrix. Conclusion: These four medicinal plant species may represent a source of alternative drugs derived from plant extracts based on folklore use and ethnobotanical knowledge from southwest Nigeria.

Keywords: extract, MRSA, antibacterial, biofilm, medicinal plants

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Authors: Pauline Bredy, Yves Schuurman, David Farrusseng

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To fulfill climate objectives, the production of synthetic e-fuels using CO₂ as a raw material appears as part of the solution. In particular, Power-to-Liquid (PtL) concept combines CO₂ with hydrogen supplied from water electrolysis, powered by renewable sources, which is currently gaining interest as it allows the production of sustainable fossil-free liquid fuels. The proposed process discussed here is an upgrading of the well-known Fischer-Tropsch synthesis. The concept deals with two cascade reactions in one pot, with first the conversion of CO₂ into CO via the reverse water gas shift (RWGS) reaction, which is then followed by the Fischer-Tropsch Synthesis (FTS). Instead of using a Fe-based catalyst, which can carry out both reactions, we have chosen the strategy to decouple the two functions (RWGS and FT) on two different catalysts within the same reactor. The FTS shall shift the equilibrium of the RWGS reaction (which alone would be limited to 15-20% of conversion at 250°C) by converting the CO into hydrocarbons. This strategy shall enable optimization of the catalyst pair and thus lower the temperature of the reaction thanks to the equilibrium shift to gain selectivity in the liquid fraction. The challenge lies in maximizing the activity of the RWGS catalyst but also in the ability of the FT catalyst to be highly selective. Methane production is the main concern as the energetic barrier of CH₄ formation is generally lower than that of the RWGS reaction, so the goal will be to minimize methane selectivity. Here we report the study of different combinations of copper-based RWGS catalysts with different cobalt-based FTS catalysts. We investigated their behaviors under mild process conditions by the use of high-throughput experimentation. Our results show that at 250°C and 20 bars, Cobalt catalysts mainly act as methanation catalysts. Indeed, CH₄ selectivity never drops under 80% despite the addition of various protomers (Nb, K, Pt, Cu) on the catalyst and its coupling with active RWGS catalysts. However, we show that the activity of the RWGS catalyst has an impact and can lead to longer hydrocarbons chains selectivities (C₂⁺) of about 10%. We studied the influence of the reduction temperature on the activity and selectivity of the tandem catalyst system. Similar selectivity and conversion were obtained at reduction temperatures between 250-400°C. This leads to the question of the active phase of the cobalt catalysts, which is currently investigated by magnetic measurements and DRIFTS. Thus, in coupling it with a more selective FT catalyst, better results are expected. This was achieved using a cobalt/iron FTS catalyst. The CH₄ selectivity dropped to 62% at 265°C, 20 bars, and a GHSV of 2500ml/h/gcat. We propose that the conditions used for the cobalt catalysts could have generated this methanation because these catalysts are known to have their best performance around 210°C in classical FTS, whereas the iron catalysts are more flexible but are also known to have an RWGS activity.

Keywords: cobalt-copper catalytic systems, CO₂-hydrogenation, Fischer-Tropsch synthesis, hydrocarbons, low-temperature process

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Authors: Hamdi Amroun, Yacine Benziani, Mehdi Ammi

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In this paper, we want to determine whether the accurate prediction of nicotine concentration can be obtained by using a network of smart objects and an e-cigarette. The approach consists of, first, the recognition of factors influencing smoking cessation such as physical activity recognition and participant’s behaviors (using both smartphone and smartwatch), then the prediction of the configuration of the e-cigarette (in terms of nicotine concentration, power, and resistance of e-cigarette). The study uses a network of commonly connected objects; a smartwatch, a smartphone, and an e-cigarette transported by the participants during an uncontrolled experiment. The data obtained from sensors carried in the three devices were trained by a Long short-term memory algorithm (LSTM). Results show that our LSTM-based model allows predicting the configuration of the e-cigarette in terms of nicotine concentration, power, and resistance with a root mean square error percentage of 12.9%, 9.15%, and 11.84%, respectively. This study can help to better control consumption of nicotine and offer an intelligent configuration of the e-cigarette to users.

Keywords: Iot, activity recognition, automatic classification, unconstrained environment

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Authors: Salma H. Abu Hafsa, A. Mendonca, B. Brehm-Stecher, A. A. Hassan, S. A. Ibrahim

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Enterococci are important inhabitants of the animal intestine and are widely used in probiotic products. A probiotic strain is expected to possess several desirable properties in order to exert beneficial effects. Therefore, the objective of this study was to isolate and characterize strains of Enterococcus sp. from chicken cecal and fecal samples to determine potential probiotic properties. Enterococci were isolated from thirty one chicken cecal and fecal samples collected from a local farm. In vitro studies were performed to assess antibacterial activity (using agar well diffusion and cell free supernatant broth technique against Salmonella enterica serotype Enteritidis), susceptibility to antibiotics (amoxycillin, cotrimoxazole, chloramphenicol, cefuroxime, ceftriaxone, ciprofloxacin, and nalidixic acid), survival in acidic conditions, resistance to bile salts, and their survival during simulated gastric juice conditions at pH 2.5. Isolates were identified using biochemical and molecular assays (API 50 CHL, and API ZYM kits followed by 16S rDNA gene sequence analysis). Two strains were identified, of which, Enteroccocus faecium was capable of inhibiting the growth of S. enteritidis and was susceptible to a wide range of antibiotics. In addition, the isolated strain exhibited significant resistance under highly acidic conditions (pH=2.5) for 8 hours and survived well in bile salt at 0.2% for 24 hours and showing ability to survive in the presence of simulated gastric juice at pH 2.5. Based on these results, the E. faecium isolate fulfills some of the criteria to be considered as a probiotic strain and therefore, could be used as a feed additive with good potential for controlling S. enteritidis in chickens. However, in vivo studies are needed to determine the safety of the strain.

Keywords: acid tolerance, antimicrobial activity, Enterococcus faecium, probiotic

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Authors: Veruska C. Dutra, Afonso R. Aquino

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Since the 1980s, global tourism activity has consolidated worldwide to become an important economic contributor, and consequently, the sociocultural and environmental impacts are starting to become evidenced. This raises the need of discussing about actions for sustainable tourism that should be linked not only to the economy, but also to the environment and social aspects. The work that is going to be presented is part of a doctoral research project in Sciences undertaken at the Sao Paulo University, Brazil. It aims to analyze whether the monitoring of the tourism sector with a focus on sustainability is applicable or not, through those indicators, put in a case study in the Jalapão State Park (JSP) conservation unit, in the state of Tocantins, Brazil. This is a study of an interdisciplinary nature that had the deductive method as its guide. We concluded that the key points of the sustainable tourism, when analyzed with the focal point in environmental indicators, are an important evaluation and quantification tool of that activity in the study locus. It displayed itself as an adequate tool for monitoring, thus decoding, the main environmental impacts that occur in tourism regions and their intensity, which is made possible through analysis, and has the objective to trace ways to prevent and correct the presented impacts.

Keywords: indicators, tourism, sustainability, Jalapão

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Authors: Giles Balbinotti, Lucas Balbinotti, Paula Hembecker

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The management of the productive process project activities, for the conception of future work and for the financial health of the companies, is an important condition in an organizational model that corroborates the management of the human aspects and their variabilities existing in the work. It is important to seek, at all levels of the organization, understanding and consequent cultural change, and so that factors associated with human aspects are considered and prioritized in the projects. In this scenario, the central question of research for this study is placed from the context of the work, in which the managers and project coordinators are inserted, as follows: How is the top management convinced, in the design stages, to take The ‘Ergonomics’ as strategy for the performance and sustainability of the business? In this perspective, this research has as general objective to analyze how the application of the management of the human aspects in a real project of productive process in the automotive industry, including the activity of the manager and coordinator of the project beyond the strategies of convincing to act in the ergonomics of design. For this, the socio-technical and ergonomic approach is adopted, given its anthropocentric premise in the sense of acting on the social system simultaneously to the technical system, besides the support of the Modapts system that measures the non-value-added times and the correlation with the Critical positions. The methodological approach adopted in this study is based on a review of the literature and the analysis of the activity of the project coordinators of an industry, including the management of human aspects in the context of work variability and the strategies applied in project activities. It was observed in the study that the loss of performance of the serial production lines reaches the important number of the order of 30%, which can make the operation with not value-added, and this loss has as one of the causes, the ergonomic problems present in the professional activity.

Keywords: human aspects in production process project, ergonomics in design, sociotechnical project management, sociotechnical, ergonomic principles, sustainability

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Authors: Kelly E. Tow, Peter Caputi, Claudia Rogge, Thomas Lee, Simon R. Knowles

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Inflammatory Bowel Disease (IBD) is a serious chronic health condition, characterised by inflammation of the gastrointestinal tract. Individuals with active IBD experience severe abdominal symptoms, which can adversely impact their physical and mental health, as well as their quality of life (QoL). Given that stress may exacerbate IBD symptoms and is frequently highlighted as a contributing factor for the development of psychological difficulties and poorer QoL, it is vital to investigate stress-management strategies aimed at improving the lives of those with IBD. The present study extends on the limited research in IBD cohorts by exploring the role of dispositional mindfulness and its impact on psychological well-being and QoL. The study examined how disease activity and dispositional mindfulness were related to psychological distress and QoL in a cohort of IBD patients. The potential role of dispositional mindfulness as a moderator between stress and anxiety, depression and QoL in these individuals was also examined. Participants included 47 patients with a clinical diagnosis of IBD. Each patient completed a series of psychological questionnaires and was assessed by a gastroenterologist to determine their disease activity levels. Correlation analyses indicated that disease activity was not significantly related to psychological distress or QoL in the sample of IBD patients. However, dispositional mindfulness was inversely related to psychological distress and positively related to QoL. Furthermore, moderation analyses demonstrated a significant interaction between stress and dispositional mindfulness on anxiety. These findings demonstrate that increased levels of dispositional mindfulness may be beneficial for individuals with IBD. Specifically, the results indicate positive links between dispositional mindfulness, general psychological well-being and QoL, and suggest that dispositional mindfulness may attenuate the negative impacts of stress on levels of anxiety in IBD patients. While further research is required to validate and expand on these findings, the current study highlights the importance of addressing psychological factors in IBD and indicates support for the use of mindfulness-based interventions for patients with the disease.

Keywords: anxiety, depression, dispositional mindfulness, inflammatory bowel disease, quality of life, stress

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Authors: Gopalu Karunakaran, Matheswaran Jagathambal, Nguyen Van Minh, Evgeny Kolesnikov, Denis Kuznetsov

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This work investigated the use of Beijerinckia indica extracellular extract for the synthesis of silver nanoparticles using AgNO₃. The formation of nanoparticles was confirmed by different methods, such as UV-Vis absorption spectroscopy, XRD, FTIR, EDX, and TEM analysis. The formation of silver nanoparticles (AgNPs) was confirmed by the change in color from light yellow to dark brown. The absorbance peak obtained at 430 nm confirmed the presence of silver nanoparticles. The XRD analysis showed the cubic crystalline phase of the synthesized nanoparticles. FTIR revealed the presence of groups that acts as stabilizing and reducing agents for silver nanoparticles formation. The synthesized silver nanoparticles were generally found to be spherical in shape with size ranging from 5 to 20 nm, as evident by TEM analysis. These nanoparticles were found to inhibit pathogenic bacterial strains. This work proved that the bacterial extract is a potential eco-friendly candidate for the synthesis of silver nanoparticles with promising antibacterial and antioxidant properties. 

Keywords: antioxidant activity, antimicrobial activity, Beijerinckia indica, characterisation, extracellular extracts, silver nanoparticles

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Authors: Bader Eddin Al Asali, N. Srinivasa Reddy

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Cities are dynamic in nature and are characterized by concentration of people, infrastructure, services and markets, which offer opportunities for production and consumption. Often growth and development in urban areas is not systematic, and is directed by number of factors like natural growth, land prices, housing availability, job locations-the central business district (CBD’s), transportation routes, distribution of resources, geographical boundaries, administrative policies, etc. One sided spatial and geographical development in cities leads to the unequal spatial distribution of population and jobs, resulting in high transportation activity. City development can be measured by the parameters such as urban size, urban form, urban shape, and urban structure. Urban Size is the city size and defined by the population of the city, and urban form is the location and size of the economic activity (CBD) over the geographical space. Urban shape is the geometrical shape of the city over which the distribution of population and economic activity occupied. And Urban Structure is the transport network within which the population and activity centers are connected by hierarchy of roads. Among the urban land-use systems transportation plays significant role and is one of the largest energy consuming sector. Transportation interaction among the land uses is measured in Passenger-Km and mean trip length, and is often used as a proxy for measurement of energy consumption in transportation sector. Among the trips generated in cities, work trips constitute more than 70 percent. Work trips are originated from the place of residence and destination to the place of employment. To understand the role of urban parameters on transportation interaction, theoretical cities of different size and urban specifications are generated through building block exercise using a specially developed interactive C++ programme and land use transportation modeling is carried. The land-use transportation modeling exercise helps in understanding the role of urban parameters and also to classify the cities for their urban form, structure, and shape. Muscat the capital city of Oman underwent rapid urbanization over the last four decades is taken as a case study for its classification. Also, a pilot survey is carried to capture urban travel characteristics. Analysis of land-use transportation modeling with field data classified Muscat as a linear city with polycentric CBD. Conclusions are drawn suggestion are given for policy making for the sustainability of Muscat City.

Keywords: land-use transportation, transportation modeling urban form, urban structure, urban rule parameters

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Authors: Aber Aboalgasm, Rupert Ward, Ruth Taylor, Jonathan Glazzard

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Teaching art by digital means is a big challenge for the majority of teachers of art and artistic design courses in primary education schools. These courses can clearly identify relationships between art, technology, and creativity in the classroom .The aim of this article is to present a modern way of teaching art, using digital tools in the art classroom in order to improve creative ability in pupils aged between 9 and 11 years; it also presents a conceptual model for creativity based on digital art. The model could be useful for pupils interested in learning drawing and using an e-drawing package, and for teachers who are interested in teaching their students modern digital art, and improving children’s creativity. This model is designed to show the strategy of teaching art through technology, in order for children to learn how to be creative. This will also help education providers to make suitable choices about which technological approaches they should choose to teach students and enhance their creative ability. It is also expected that use of this model will help to develop social interactive qualities that may improve intellectual ability.

Keywords: digital tools, motivation, creative activity, education

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Authors: Renu Kumari, Jyotsna Dutta Majumdar

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In the present study, composite coating consisting of hydroxyapatite (HA) + 50 wt% TiO2 has been developed on Ti-6Al-4V substrate by plasma spray deposition technique. Followed by plasma spray deposition, detailed surface roughness and microstructural characterization were carried out by using optical profilometer and scanning electron microscopy (SEM), respectively. The composition and phase analysis were carried out by energy-dispersive X-ray spectroscopy analysis, and X-ray diffraction (XRD) technique, respectively. The bio-activity behavior of the uncoated and coated samples was also compared by dipping test in Hank’s solution. The average surface roughness of the coating was 10 µm (as compared to 0.5 µm of as-received Ti-6Al-4V substrate) with the presence of porosities. The microstructure of the coating was found to be continuous with the presence of solidified splats. A detailed XRD analysis shows phase transformation of TiO2 from anatase to rutile, decomposition of hydroxyapatite, and formation of CaTiO3 phase. Standard dipping test confirmed a faster kinetics of deposition of calcium phosphate in the coated HA+50% wt.% TiO2 surface as compared to the as-received substrate.

Keywords: titanium, plasma spraying, microstructure, bio-activity, TiO2, hydroxyapatite

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Authors: Mei Chen, Yingping Hou, Michelle Hao, Soheil Aghamohammadzadeh, Esteban Terzo, Roger Clark, Vijay Modur

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Background: Recessive Dystrophic Epidermolysis Bullosa (RDEB) is a genetic skin condition characterized by skin tearing and unremitting blistering upon minimal trauma. Repeated blistering, fibrosis, and scarring lead to aggressive squamous cell carcinoma later in life. RDEB is caused by mutations in the COL7A1 gene encoding collagen type VII (C7), the major component of anchoring fibrils mediating epidermis-dermis adherence. Nonsense mutations in the COL7A1 gene of a subset of RDEB patients leads to premature termination codons (PTC). Similarly, most Junctional Epidermolysis Bullosa (JEB) cases are caused by nonsense mutations in the LAMB3 gene encoding the β3 subunit of laminin 332. Currently, there is an unmet need for the treatment of RDEB and JEB. Zikani Therapeutics has discovered an array of macrocyclic compounds with ring structures similar to macrolide antibiotics that can facilitate readthrough activity of nonsense mutations in the COL7A1 and LAMB3 genes by acting as Ribosome Modulating Agents (RMAs). The medicinal chemistry synthetic advancements of these macrocyclic compounds have allowed targeting the human ribosome while preserving the structural elements responsible for the safety and pharmacokinetic profile of clinically used macrolide antibiotics. Methods: C7 expression was used as a measure of readthrough activity by immunoblot assays in two primary human fibroblasts from RDEB patients (R578X/R578X and R163X/R1683X-COL7A1). Similarly, immunoblot assays in C325X/c.629-12T > A-LAMB3 keratinocytes were used to measure readthrough activity for JEB. The relative readthrough activity of each compound was measured relative to Gentamicin. An imaging-based fibroblast migration assay was used as an assessment of C7 functionality in RDEB-fibroblasts over 16-20 hrs. The incubation period for the above experiments was 48 hrs for RDEB fibroblasts and 72 hours for JEB keratinocytes. Results: 9 RMAs demonstrated increased protein expression in both patient RDEB fibroblasts. The highest readthrough activity at tested concentrations without cytotoxicities increased protein expression up to 179% of Gentamicin (400 µg/ml), with favored readthrough activity in R163X/R1683X-COL7A1 fibroblasts. Concurrent with protein expression, fibroblast hypermotility phenotype observed in RDEB was rescued by reducing motility by ~35% to WT levels (the same level as 690 µM Gentamicin treated cells). Laminin β3 expression was also shown to be increased by 6 RMAs in keratinocytes to 33-83% of (400 µg/ml) Gentamicin. Conclusions: To date, 9 RMAs have been identified that enhance the expression of functional C7 in a mutation-dependent manner in two different RDEB patient fibroblast backgrounds (R578X/R578X and R163X/R1683X-COL7A1). A further 6 RMAs have been identified that enhance the readthrough of C325X-LAMB3 in JEB patient keratinocytes. Based on the clinical trial conducted by us with topical gentamycin in 2017, Zikani’s RMAs achieve clinically significant levels of read-through for the treatment of recessive dystrophic and Junctional Epidermolysis Bullosa.

Keywords: epidermolysis bullosa, nonsense mutation, readthrough, ribosome modulation

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