Search results for: structure and texture of rocks
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Search results for: structure and texture of rocks

553 A Holistic View of Microbial Community Dynamics during a Toxic Harmful Algal Bloom

Authors: Shi-Bo Feng, Sheng-Jie Zhang, Jin Zhou

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The relationship between microbial diversity and algal bloom has received considerable attention for decades. Microbes undoubtedly affect annual bloom events and impact the physiology of both partners, as well as shape ecosystem diversity. However, knowledge about interactions and network correlations among broader-spectrum microbes that lead to the dynamics in a complete bloom cycle are limited. In this study, pyrosequencing and network approaches simultaneously assessed the associate patterns among bacteria, archaea, and microeukaryotes in surface water and sediments in response to a natural dinoflagellate (Alexandrium sp.) bloom. In surface water, among the bacterial community, Gamma-Proteobacteria and Bacteroidetes dominated in the initial bloom stage, while Alpha-Proteobacteria, Cyanobacteria, and Actinobacteria become the most abundant taxa during the post-stage. In the archaea biosphere, it clustered predominantly with Methanogenic members in the early pre-bloom period while the majority of species identified in the later-bloom stage were ammonia-oxidizing archaea and Halobacteriales. In eukaryotes, dinoflagellate (Alexandrium sp.) was dominated in the onset stage, whereas multiply species (such as microzooplankton, diatom, green algae, and rotifera) coexistence in bloom collapse stag. In sediments, the microbial species biomass and richness are much higher than the water body. Only Flavobacteriales and Rhodobacterales showed a slight response to bloom stages. Unlike the bacteria, there are small fluctuations of archaeal and eukaryotic structure in the sediment. The network analyses among the inter-specific associations show that bacteria (Alteromonadaceae, Oceanospirillaceae, Cryomorphaceae, and Piscirickettsiaceae) and some zooplankton (Mediophyceae, Mamiellophyceae, Dictyochophyceae and Trebouxiophyceae) have a stronger impact on the structuring of phytoplankton communities than archaeal effects. The changes in population were also significantly shaped by water temperature and substrate availability (N & P resources). The results suggest that clades are specialized at different time-periods and that the pre-bloom succession was mainly a bottom-up controlled, and late-bloom period was controlled by top-down patterns. Additionally, phytoplankton and prokaryotic communities correlated better with each other, which indicate interactions among microorganisms are critical in controlling plankton dynamics and fates. Our results supplied a wider view (temporal and spatial scales) to understand the microbial ecological responses and their network association during algal blooming. It gives us a potential multidisciplinary explanation for algal-microbe interaction and helps us beyond the traditional view linked to patterns of algal bloom initiation, development, decline, and biogeochemistry.

Keywords: microbial community, harmful algal bloom, ecological process, network

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552 Effect of Vitrification on Embryos Euploidy Obtained from Thawed Oocytes

Authors: Natalia Buderatskaya, Igor Ilyin, Julia Gontar, Sergey Lavrynenko, Olga Parnitskaya, Ekaterina Ilyina, Eduard Kapustin, Yana Lakhno

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Introduction: It is known that cryopreservation of oocytes has peculiar features due to the complex structure of the oocyte. One of the most important features is that mature oocytes contain meiotic division spindle which is very sensitive even to the slightest variation in temperature. Thus, the main objective of this study is to analyse the resulting euploid embryos obtained from thawed oocytes in comparison with the data of preimplantation genetic screening (PGS) in fresh embryo cycles. Material and Methods: The study was conducted at 'Medical Centre IGR' from January to July 2016. Data were analysed for 908 donor oocytes obtained in 67 cycles of assisted reproductive technologies (ART), of which 693 oocytes were used in the 51 'fresh' cycles (group A), and 215 oocytes - 16 ART programs with vitrification female gametes (group B). The average age of donors in the groups match 27.3±2.9 and 27.8±6.6 years. Stimulation of superovulation was conducted the standard way. Vitrification was performed in 1-2 hours after transvaginal puncture and thawing of oocytes were carried out in accordance with the standard protocol of Cryotech (Japan). Manipulation ICSI was performed 4-5 hours after transvaginal follicle puncture for fresh oocytes, or after defrosting - for vitrified female gametes. For the PGS, an embryonic biopsy was done on the third or on the fifth day after fertilization. Diagnostic procedures were performed using fluorescence in situ hybridization with the study of such chromosomes as 13, 16, 18, 21, 22, X, Y. Only morphologically quality blastocysts were used for the transfer, the estimation of which corresponded to the Gardner criteria. The statistical hypotheses were done using the criteria t, x^2 at a significance levels p<0.05, p<0.01, p<0.001. Results: The mean number of mature oocytes per cycle in group A was 13.58±6.65 and in group B - 13.44±6.68 oocytes for patient. The survival of oocytes after thawing totaled 95.3% (n=205), which indicates a highly effective quality of performed vitrification. The proportion of zygotes in the group A corresponded to 91.1%(n=631), in the group B – 80.5%(n=165), which shows statistically significant difference between the groups (p<0.001) and explained by non-viable oocytes elimination after vitrification. This is confirmed by the fact that on the fifth day of embryos development a statistically significant difference in the number of blastocysts was absent (p>0.05), and constituted respectively 61.6%(n=389) and 63.0%(n=104) in the groups. For the PGS performing 250 embryos analyzed in the group A and 72 embryos - in the group B. The results showed that euploidy in the studied chromosomes were 40.0%(n=100) embryos in the group A and 41.7% (n=30) - in the group B, which shows no statistical significant difference (p>0.05). The indicators of clinical pregnancies in the groups amounted to 64.7% (22 pregnancies per 34 embryo transfers) and 61.5% (8 pregnancies per 13 embryo transfers) respectively, and also had no significant difference between the groups (p>0.05). Conclusions: The results showed that the vitrification does not affect the resulting euploid embryos in assisted reproductive technologies and are not reflected in their morphological characteristics in ART programs.

Keywords: euploid embryos, preimplantation genetic screening, thawing oocytes, vitrification

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551 Study of Chemical State Analysis of Rubidium Compounds in Lα, Lβ₁, Lβ₃,₄ and Lγ₂,₃ X-Ray Emission Lines with Wavelength Dispersive X-Ray Fluorescence Spectrometer

Authors: Harpreet Singh Kainth

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Rubidium salts have been commonly used as an electrolyte to improve the efficiency cycle of Li-ion batteries. In recent years, it has been implemented into the large scale for further technological advances to improve the performance rate and better cyclability in the batteries. X-ray absorption spectroscopy (XAS) is a powerful tool for obtaining the information in the electronic structure which involves the chemical state analysis in the active materials used in the batteries. However, this technique is not well suited for the industrial applications because it needs a synchrotron X-ray source and special sample file for in-situ measurements. In contrast to this, conventional wavelength dispersive X-ray fluorescence (WDXRF) spectrometer is nondestructive technique used to study the chemical shift in all transitions (K, L, M, …) and does not require any special pre-preparation planning. In the present work, the fluorescent Lα, Lβ₁ , Lβ₃,₄ and Lγ₂,₃ X-ray spectra of rubidium in different chemical forms (Rb₂CO₃ , RbCl, RbBr, and RbI) have been measured first time with high resolution wavelength dispersive X-ray fluorescence (WDXRF) spectrometer (Model: S8 TIGER, Bruker, Germany), equipped with an Rh anode X-ray tube (4-kW, 60 kV and 170 mA). In ₃₇Rb compounds, the measured energy shifts are in the range (-0.45 to - 1.71) eV for Lα X-ray peak, (0.02 to 0.21) eV for Lβ₁ , (0.04 to 0.21) eV for Lβ₃ , (0.15 to 0.43) eV for Lβ₄ and (0.22 to 0.75) eV for Lγ₂,₃ X-ray emission lines. The chemical shifts in rubidium compounds have been measured by considering Rb₂CO₃ compounds taking as a standard reference. A Voigt function is used to determine the central peak position of all compounds. Both positive and negative shifts have been observed in L shell emission lines. In Lα X-ray emission lines, all compounds show negative shift while in Lβ₁, Lβ₃,₄, and Lγ₂,₃ X-ray emission lines, all compounds show a positive shift. These positive and negative shifts result increase or decrease in X-ray energy shifts. It looks like that ligands attached with central metal atom attract or repel the electrons towards or away from the parent nucleus. This pulling and pushing character of rubidium affects the central peak position of the compounds which causes a chemical shift. To understand the chemical effect more briefly, factors like electro-negativity, line intensity ratio, effective charge and bond length are responsible for the chemical state analysis in rubidium compounds. The effective charge has been calculated from Suchet and Pauling method while the line intensity ratio has been calculated by calculating the area under the relevant emission peak. In the present work, it has been observed that electro-negativity, effective charge and intensity ratio (Lβ₁/Lα, Lβ₃,₄/Lα and Lγ₂,₃/Lα) are inversely proportional to the chemical shift (RbCl > RbBr > RbI), while bond length has been found directly proportional to the chemical shift (RbI > RbBr > RbCl).

Keywords: chemical shift in L emission lines, bond length, electro-negativity, effective charge, intensity ratio, Rubidium compounds, WDXRF spectrometer

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550 Effects of Dietary Polyunsaturated Fatty Acids and Beta Glucan on Maturity, Immunity and Fry Quality of Pabdah Catfish, Ompok pabda

Authors: Zakir Hossain, Md. Saddam Hossain

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A nutritionally balanced diet and selection of appropriate species are important criteria in aquaculture. The present study was conducted to evaluate the effects of polyunsaturated fatty acids (PUFAs) and beta glucan containing diet on growth performance, feed utilization, maturation, immunity, early embryonic and larval development of endangered Pabdah catfish, Ompok pabda. In this study, squid extracted lipids and mushroom powder were used as the source of PUFAs and beta glucan, respectively, and formulated two isonitrogenous diets such as basal or control (CON) diet and treated (PBG) diet with maintaining 30% protein levels. During the study period, similar physicochemical conditions of water such as temperature, pH, and dissolved oxygen (DO) were 26.5±2 °C, 7.4±0.2, and 6.7±0.5 ppm, respectively in each cistern. The results showed that final mean body weight, final mean length gain, food conversion ratio (FCR), specific growth rate (SGR), food conversion efficiency (%), hepatosomatic index (HSI), kidney index (KI), and viscerosomatic index (VSI) were significantly (P<0.01 and P<0.05) higher in fish fed the PBG diet than that of fish fed the CON diet. The length-weight relationship and relative condition factor (K) of O. pabda were significantly (P<0.05) affected by the PBG diet. The gonadosomatic index (GSI), sperm viability, blood serum calcium ion concentrations (Ca²⁺), and vitellogenin level were significantly (P<0.05) higher in fish fed the PBG diet than that of fish fed the CON diet; which was used to the indication of fish maturation. During the spawning season, lipid granules and normal morphological structure were observed in the treated fish liver, whereas fewer lipid granules of liver were observed in the control group. Based on the immunity and stress resistance-related parameters such as hematological indices, antioxidant activity, lysozyme level, respiratory burst activity, blood reactive oxygen species (ROS), complement activity (ACH50 assay), specific IgM, brain AChE, plasma PGOT, and PGPT enzyme activity were significantly (P<0.01 and P<0.05) higher in fish fed the PBG diet than that of fish fed the CON diet. The fecundity, fertilization rate (92.23±2.69%), hatching rate (87.43±2.17 %) and survival (76.62±0.82%) of offspring were significantly higher (P˂0.05) in the PBG diet than in the control. Consequently, early embryonic and larval development was better in PBG treated group than in the control. Therefore, the present study showed that the polyunsaturated fatty acids (PUFAs) and beta glucan enriched experimental diet were more effective and achieved better growth, feed utilization, maturation, immunity, and spawning performances of O. pabda.

Keywords: polyunsaturated fatty acids, beta glucan, maturity, immunity, catfish

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549 Influence of Gamma-Radiation Dosimetric Characteristics on the Stability of the Persistent Organic Pollutants

Authors: Tatiana V. Melnikova, Lyudmila P. Polyakova, Alla A. Oudalova

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As a result of environmental pollution, the production of agriculture and foodstuffs inevitably contain residual amounts of Persistent Organic Pollutants (POP). The special attention must be given to organic pollutants, including various organochlorinated pesticides (OCP). Among priorities, OCP is DDT (and its metabolite DDE), alfa-HCH, gamma-HCH (lindane). The control of these substances spends proceeding from requirements of sanitary norms and rules. During too time often is lost sight of that the primary product can pass technological processing (in particular irradiation treatment) as a result of which transformation of physicochemical forms of initial polluting substances is possible. The goal of the present work was to study the OCP radiation degradation at a various gamma-radiation dosimetric characteristics. The problems posed for goal achievement: to evaluate the content of the priority of OCPs in food; study the character the degradation of OCP in model solutions (with micro concentrations commensurate with the real content of their agricultural and food products) depending upon dosimetric characteristics of gamma-radiation. Qualitative and quantitative analysis of OCP in food and model solutions by gas chromatograph Varian 3400 (Varian, Inc. (USA)); chromatography-mass spectrometer Varian Saturn 4D (Varian, Inc. (USA)) was carried out. The solutions of DDT, DDE, alpha- and gamma- isomer HCH (0.01, 0.1, 1 ppm) were irradiated on "Issledovatel" (60Co) and "Luch - 1" (60Co) installations at a dose 10 kGy with a variation of dose rate from 0.0083 up to 2.33 kGy/sec. It was established experimentally that OCP residual concentration in individual samples of food products (fish, milk, cereal crops, meat, butter) are evaluated as 10-1-10-4 mg/kg, the value of which depends on the factor-sensations territory and natural migration processes. The results were used in the preparation of model solutions OCP. The dependence of a degradation extent of OCP from a dose rate gamma-irradiation has complex nature. According to our data at a dose 10 kGy, the degradation extent of OCP at first increase passes through a maximum (over the range 0.23 – 0.43 Gy/sec), and then decrease with the magnification of a dose rate. The character of the dependence of a degradation extent of OCP from a dose rate is kept for various OCP, in polar and nonpolar solvents and does not vary at the change of concentration of the initial substance. Also in work conditions of the maximal radiochemical yield of OCP which were observed at having been certain: influence of gamma radiation with a dose 10 kGy, in a range of doses rate 0.23 – 0.43 Gy/sec; concentration initial OCP 1 ppm; use of solvent - 2-propanol after preliminary removal of oxygen. Based on, that at studying model solutions of OCP has been established that the degradation extent of pesticides and qualitative structure of OCP radiolysis products depend on a dose rate, has been decided to continue researches radiochemical transformations OCP into foodstuffs at various of doses rate.

Keywords: degradation extent, dosimetric characteristics, gamma-radiation, organochlorinated pesticides, persistent organic pollutants

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548 Understanding Project Failures in Construction: The Critical Impact of Financial Capacity

Authors: Nnadi Ezekiel Oluwaseun Ejiofor

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This research investigates the effects of poor cost estimation, material cost variations, and payment punctuality on the financial health and execution of construction projects in Nigeria. To achieve the objectives of the study, a quantitative research approach was employed, and data was gathered through an online survey of 74 construction industry professionals consisting of quantity surveyors, contractors, and other professionals. The study surveyed input on cost estimation errors, price fluctuations, and payment delays, among other factors. The responses of the respondents were analyzed using a five-point Likert scale and the Relative Importance Index (RII). The findings demonstrated that the errors in cost estimating in the Bill of Quantity (BOQ) have a high degree of negative impact on the reputation and image of the participants in the projects. The greatest effect was experienced on the likelihood of obtaining future endeavors for contractors (mean value = 3.42), followed by the likelihood of obtaining new commissions by quantity surveyors (mean value = 3.40). The level of inaccuracy in costing that undershoots exposes them to risks was most serious in terms of easement of construction and effects of shortage of funds to pursue bankruptcy (hence fears of mean value = 3.78). There was also considerable financial damage as a result of cost underestimation, whereby contractors suffered the worst loss in profit (mean value = 3.88). Every expense comes with its own peculiar risk and uncertainty. Pressure on the cost of materials and every other expense attributed to the building and completion of a structure adds risks to the performance figures of a project. The greatest weight (mean importance score = 4.92) was attributed to issues like market inflation in building materials, while the second greatest weight (mean importance score = 4.76) was due to increased transportation charges. On the other hand, delays in payments arising from issues of the clients like poor availability of funds (RII=0.71) and contracting issues such as disagreements on the valuation of works done (RII=0.72) or other reasons were also found to lead to project delays and additional costs. The results affirm the importance of proper cost estimation on the health of organization finances and project risks and finishes within set time limits. As for the suggestions, it is proposed to progress on the methods of costing, engender better communications with the stakeholders, and manage the delays by way of contracting and financial control. This study enhances the existing literature on construction project management by suggesting ways to deal with adverse cost inaccuracies and availability of materials due to delays in payments which, if addressed, would greatly improve the economic performance of the construction business.

Keywords: cost estimation, construction project management, material price fluctuations, payment delays, financial impact

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547 Articulating the Colonial Relation, a Conversation between Afropessimism and Anti-Colonialism

Authors: Thomas Compton

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As Decolonialism becomes an important topic in Political Theory, the rupture between the colonized and the colonist relation has lost attention. Focusing on the anti-colonial activist Madhi Amel, we shall consider his attention to the permanence of the colonial relation and how it preempts Frank Wilderson’s formulation of (white) culturally necessary Anti-Black violence. Both projects draw attention away from empirical accounts of oppression, instead focusing on the structural relation which precipitates them. As Amel says that we should stop thinking of the ‘underdeveloped’ as beyond the colonial relation, Wilderson says we should stop think of the Black rights that have surpassed the role of the slave. However, Amel moves beyond his idol Althusser’s Structuralism toward a formulation of the colonial relation as source of domination. Our analysis will take a Lacanian turn in considering how this non-relation was formulated as a relation how this space of negativity became a ideological opportunity for Colonial domination. Wilderson’s work shall problematise this as we conclude with his criticisms of Structural accounts for the failure to consider how Black social death exists as more than necessity but a cite of white desire. Amel, a Lebanese activist and scholar (re)discovered by Hicham Safieddine, argues colonialism is more than the theft of land, but instead a privatization of collective property and form of investment which (re)produces the status of the capitalist in spaces ‘outside’ the market. Although Amel was a true Marxist-Leninsist, who exposited the economic determinacy of the Colonial Mode of Production, we are reading this account through Alenka Zupančič’s reformulation of the ‘invisible hand job of the market’. Amel points to the signifier ‘underdeveloped’ as buttressed on a pre-colonial epistemic break, as the Western investor (debt collector) sees the (post?) colony narcissistic image. However, the colony can never become site of class conflict, as the workers are not unified but existing between two countries. In industry, they are paid in Colonial subjectivisation, the promise of market (self)pleasure, at home, they are refugees. They are not, as Wilderson states, in the permanent social death of the slave, but they are less than the white worker. This is formulated as citizen (white), non-citizen (colonized), anti-citizen (Black/slave). Here we may also think of how indentured Indians were used as instruments of colonial violence. Wilderson’s aphorism “there is no analogy to anti-Black violence” lays bare his fundamental opposition between colonial and specifically anti-Black violence. It is not only that the debt collector, landowner, or other owners of production pleasures themselves as if their hand is invisible. The absolute negativity between colony and colonized provides a new frontier for desire, the development of a colonial mode of production. An invention inside the colonial structure that is generative of class substitution. We shall explore how Amel ignores the role of the slave but how Wilderson forecloses the history African anti-colonial.

Keywords: afropessimism, fanon, marxism, postcolonialism

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546 Improvement of Activity of β-galactosidase from Kluyveromyces lactis via Immobilization on Polyethylenimine-Chitosan

Authors: Carlos A. C. G. Neto, Natan C. G. e Silva , Thaís de O. Costa, Luciana R. B. Gonçalves, Maria V. P. Rocha

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β-galactosidases (E.C. 3.2.1.23) are enzymes that have attracted by catalyzing the hydrolysis of lactose and in producing galacto-oligosaccharides by favoring transgalactosylation reactions. These enzymes, when immobilized, can have some enzymatic characteristics substantially improved, and the coating of supports with multifunctional polymers is a promising alternative to enhance the stability of the biocatalysts, among which polyethylenimine (PEI) stands out. PEI has certain properties, such as being a flexible polymer that suits the structure of the enzyme, giving greater stability, especially for multimeric enzymes such as β-galactosidases. Besides that, protects them from environmental variations. The use of chitosan support coated with PEI could improve the catalytic efficiency of β-galactosidase from Kluyveromyces lactis in the transgalactosylation reaction for the production of prebiotics, such as lactulose since this strain is more effective in the hydrolysis reaction. In this context, the aim of the present work was first to develop biocatalysts of β-galactosidase from K. lactis immobilized on chitosan-coated with PEI, determining the immobilization parameters, its operational and thermal stability, and then to apply it in hydrolysis and transgalactolisation reactions to produce lactulose using whey as a substrate. The immobilization of β-galactosidase in chitosan previously functionalized with 0.8% (v/v) glutaraldehyde and then coated with 10% (w/v) PEI solution was evaluated using an enzymatic load of 10 mg protein per gram support. Subsequently, the hydrolysis and transgalactosylation reactions were conducted at 50 °C, 120 RPM for 20 minutes, using whey supplemented with fructose at a ratio of 1:2 lactose/fructose, totaling 200 g/L. Operational stability studies were performed in the same conditions for 10 cycles. Thermal stabilities of biocatalysts were conducted at 50 ºC in 50 mM phosphate buffer, pH 6.6 with 0.1 mM MnCl2. The biocatalyst whose support was coated was named CHI_GLU_PEI_GAL, and the one that was not coated was named CHI_GLU_GAL. The coating of the support with PEI considerably improved the parameters of immobilization. The immobilization yield increased from 56.53% to 97.45%, biocatalyst activity from 38.93 U/g to 95.26 U/g and the efficiency from 3.51% to 6.0% for uncoated and coated support, respectively. The biocatalyst CHI_GLU_PEI_GAL was better than CHI_GLU_GAL in the hydrolysis of lactose and production of lactulose, converting 97.05% of lactose at 5 min of reaction and producing 7.60 g/L lactulose in the same time interval. QUI_GLU_PEI_GAL biocatalyst was stable in the hydrolysis reactions of lactose during the 10 cycles evaluated, converting 73.45% lactose even after the tenth cycle, and in the lactulose production was stable until the fifth cycle evaluated, producing 10.95 g/L lactulose. However, the thermal stability of CHI_GLU_GAL biocatalyst was superior, with a half-life time 6 times higher, probably because the enzyme was immobilized by covalent bonding, which is stronger than adsorption (CHI_GLU_PEI_GAL). Therefore, the strategy of coating the supports with PEI has proven to be effective for the immobilization of β-galactosidase from K. lactis, considerably improving the immobilization parameters, as well as, the catalytic action of the enzyme. Besides that, this process can be economically viable due to the use of an industrial residue as a substrate.

Keywords: β-galactosidase, immobilization, kluyveromyces lactis, lactulose, polyethylenimine, transgalactosylation reaction, whey

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545 Development of Bilayer Coating System for Mitigating Corrosion of Offshore Wind Turbines

Authors: Adamantini Loukodimou, David Weston, Shiladitya Paul

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Offshore structures are subjected to harsh environments. It is documented that carbon steel needs protection from corrosion. The combined effect of UV radiation, seawater splash, and fluctuating temperatures diminish the integrity of these structures. In addition, the possibility of damage caused by floating ice, seaborne debris, and maintenance boats make them even more vulnerable. Their inspection and maintenance when far out in the sea are difficult, risky, and expensive. The most known method of mitigating corrosion of offshore structures is the use of cathodic protection. There are several zones in an offshore wind turbine. In the atmospheric zone, due to the lack of a continuous electrolyte (seawater) layer between the structure and the anode at all times, this method proves inefficient. Thus, the use of protective coatings becomes indispensable. This research focuses on the atmospheric zone. The conversion of commercially available and conventional paint (epoxy) system to an autonomous self-healing paint system via the addition of suitable encapsulated healing agents and catalyst is investigated in this work. These coating systems, which can self-heal when damaged, can provide a cost-effective engineering solution to corrosion and related problems. When the damage of the paint coating occurs, the microcapsules are designed to rupture and release the self-healing liquid (monomer), which then will react in the presence of the catalyst and solidify (polymerization), resulting in healing. The catalyst should be compatible with the system because otherwise, the self-healing process will not occur. The carbon steel substrate will be exposed to a corrosive environment, so the use of a sacrificial layer of Zn is also investigated. More specifically, the first layer of this new coating system will be TSZA (Thermally Sprayed Zn85/Al15) and will be applied on carbon steel samples with dimensions 100 x 150 mm after being blasted with alumina (size F24) as part of the surface preparation. Based on the literature, it corrodes readily, so one additional paint layer enriched with microcapsules will be added. Also, the reaction and the curing time are of high importance in order for this bilayer system of coating to work successfully. For the first experiments, polystyrene microcapsules loaded with 3-octanoyltio-1-propyltriethoxysilane were conducted. Electrochemical experiments such as Electrochemical Impedance Spectroscopy (EIS) confirmed the corrosion inhibiting properties of the silane. The diameter of the microcapsules was about 150-200 microns. Further experiments were conducted with different reagents and methods in order to obtain diameters of about 50 microns, and their self-healing properties were tested in synthetic seawater using electrochemical techniques. The use of combined paint/electrodeposited coatings allows for further novel development of composite coating systems. The potential for the application of these coatings in offshore structures will be discussed.

Keywords: corrosion mitigation, microcapsules, offshore wind turbines, self-healing

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544 Predicting Long-Term Performance of Concrete under Sulfate Attack

Authors: Elakneswaran Yogarajah, Toyoharu Nawa, Eiji Owaki

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Cement-based materials have been using in various reinforced concrete structural components as well as in nuclear waste repositories. The sulfate attack has been an environmental issue for cement-based materials exposed to sulfate bearing groundwater or soils, and it plays an important role in the durability of concrete structures. The reaction between penetrating sulfate ions and cement hydrates can result in swelling, spalling and cracking of cement matrix in concrete. These processes induce a reduction of mechanical properties and a decrease of service life of an affected structure. It has been identified that the precipitation of secondary sulfate bearing phases such as ettringite, gypsum, and thaumasite can cause the damage. Furthermore, crystallization of soluble salts such as sodium sulfate crystals induces degradation due to formation and phase changes. Crystallization of mirabilite (Na₂SO₄:10H₂O) and thenardite (Na₂SO₄) or their phase changes (mirabilite to thenardite or vice versa) due to temperature or sodium sulfate concentration do not involve any chemical interaction with cement hydrates. Over the past couple of decades, an intensive work has been carried out on sulfate attack in cement-based materials. However, there are several uncertainties still exist regarding the mechanism for the damage of concrete in sulfate environments. In this study, modelling work has been conducted to investigate the chemical degradation of cementitious materials in various sulfate environments. Both internal and external sulfate attack are considered for the simulation. In the internal sulfate attack, hydrate assemblage and pore solution chemistry of co-hydrating Portland cement (PC) and slag mixing with sodium sulfate solution are calculated to determine the degradation of the PC and slag-blended cementitious materials. Pitzer interactions coefficients were used to calculate the activity coefficients of solution chemistry at high ionic strength. The deterioration mechanism of co-hydrating cementitious materials with 25% of Na₂SO₄ by weight is the formation of mirabilite crystals and ettringite. Their formation strongly depends on sodium sulfate concentration and temperature. For the external sulfate attack, the deterioration of various types of cementitious materials under external sulfate ingress is simulated through reactive transport model. The reactive transport model is verified with experimental data in terms of phase assemblage of various cementitious materials with spatial distribution for different sulfate solution. Finally, the reactive transport model is used to predict the long-term performance of cementitious materials exposed to 10% of Na₂SO₄ for 1000 years. The dissolution of cement hydrates and secondary formation of sulfate-bearing products mainly ettringite are the dominant degradation mechanisms, but not the sodium sulfate crystallization.

Keywords: thermodynamic calculations, reactive transport, radioactive waste disposal, PHREEQC

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543 Polarimetric Study of System Gelatin / Carboxymethylcellulose in the Food Field

Authors: Sihem Bazid, Meriem El Kolli, Aicha Medjahed

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Proteins and polysaccharides are the two types of biopolymers most frequently used in the food industry to control the mechanical properties and structural stability and organoleptic properties of the products. The textural and structural properties of these two types of blend polymers depend on their interaction and their ability to form organized structures. From an industrial point of view, a better understanding of mixtures protein / polysaccharide is an important issue since they are already heavily involved in processed food. It is in this context that we have chosen to work on a model system composed of a fibrous protein mixture (gelatin)/anionic polysaccharide (sodium carboxymethylcellulose). Gelatin, one of the most popular biopolymers, is widely used in food, pharmaceutical, cosmetic and photographic applications, because of its unique functional and technological properties. Sodium Carboxymethylcellulose (NaCMC) is an anionic linear polysaccharide derived from cellulose. It is an important industrial polymer with a wide range of applications. The functional properties of this anionic polysaccharide can be modified by the presence of proteins with which it might interact. Another factor may also manage the interaction of protein-polysaccharide mixtures is the triple helix of the gelatin. Its complex synthesis method results in an extracellular assembly containing several levels. Collagen can be in a soluble state or associate into fibrils, which can associate in fiber. Each level corresponds to an organization recognized by the cellular and metabolic system. Gelatin allows this approach, the formation of gelatin gel has triple helical folding of denatured collagen chains, this gel has been the subject of numerous studies, and it is now known that the properties depend only on the rate of triple helices forming the network. Chemical modification of this system is quite controlled. Observe the dynamics of the triple helix may be relevant in understanding the interactions involved in protein-polysaccharides mixtures. Gelatin is central to any industrial process, understand and analyze the molecular dynamics induced by the triple helix in the transitions gelatin, can have great economic importance in all fields and especially the food. The goal is to understand the possible mechanisms involved depending on the nature of the mixtures obtained. From a fundamental point of view, it is clear that the protective effect of NaCMC on gelatin and conformational changes of the α helix are strongly influenced by the nature of the medium. Our goal is to minimize the maximum the α helix structure changes to maintain more stable gelatin and protect against denaturation that occurs during such conversion processes in the food industry. In order to study the nature of interactions and assess the properties of mixtures, polarimetry was used to monitor the optical parameters and to assess the rate of helicity gelatin.

Keywords: gelatin, sodium carboxymethylcellulose, interaction gelatin-NaCMC, the rate of helicity, polarimetry

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542 Tracing a Timber Breakthrough: A Qualitative Study of the Introduction of Cross-Laminated-Timber to the Student Housing Market in Norway

Authors: Marius Nygaard, Ona Flindall

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The Palisaden student housing project was completed in August 2013 and was, with its eight floors, Norway’s tallest timber building at the time of completion. It was the first time cross-laminated-timber (CLT) was utilized at this scale in Norway. The project was the result of a concerted effort by a newly formed management company to establish CLT as a sustainable and financially competitive alternative to conventional steel and concrete systems. The introduction of CLT onto the student housing market proved so successful that by 2017 more than 4000 individual student residences will have been built using the same model of development and construction. The aim of this paper is to identify the key factors that enabled this breakthrough for CLT. It is based on an in-depth study of a series of housing projects and the role of the management company who both instigated and enabled this shift of CLT from the margin to the mainstream. Specifically, it will look at how a new building system was integrated into a marketing strategy that identified a market potential within the existing structure of the construction industry and within the economic restrictions inherent to student housing in Norway. It will show how a key player established a project model that changed both the patterns of cooperation and the information basis for decisions. Based on qualitative semi-structured interviews with managers, contractors and the interdisciplinary teams of consultants (architects, structural engineers, acoustical experts etc.) this paper will trace the introduction, expansion and evolution of CLT-based building systems in the student housing market. It will show how the project management firm’s position in the value chain enabled them to function both as a liaison between contractor and client, and between contractor and producer. A position that allowed them to improve the flow of information. This ensured that CLT was handled on equal terms to other structural solutions in the project specifications, enabling realistic pricing and risk evaluation. Secondly, this paper will describe and discuss how the project management firm established and interacted with a growing network of contractors, architects and engineers to pool expertise and broaden the knowledge base across Norway’s regional markets. Finally, it will examine the role of the client, the building typology, and the industrial and technological factors in achieving this breakthrough for CLT in the construction industry. This paper gives an in-depth view of the progression of a single case rather than a broad description of the state of the art of large-scale timber building in Norway. However, this type of study may offer insights that are important to the understanding not only of specific markets but also of how new technologies should be introduced in big and well-established industries.

Keywords: cross-laminated-timber (CLT), industry breakthrough, student housing, timber market

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541 An Investigation on the Sandwich Panels with Flexible and Toughened Adhesives under Flexural Loading

Authors: Emre Kara, Şura Karakuzu, Ahmet Fatih Geylan, Metehan Demir, Kadir Koç, Halil Aykul

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The material selection in the design of the sandwich structures is very crucial aspect because of the positive or negative influences of the base materials to the mechanical properties of the entire panel. In the literature, it was presented that the selection of the skin and core materials plays very important role on the behavior of the sandwich. Beside this, the use of the correct adhesive can make the whole structure to show better mechanical results and behavior. By this way, the sandwich structures realized in the study were obtained with the combination of aluminum foam core and three different glass fiber reinforced polymer (GFRP) skins using two different commercial adhesives which are based on flexible polyurethane and toughened epoxy. The static and dynamic tests were already applied on the sandwiches with different types of adhesives. In the present work, the static three-point bending tests were performed on the sandwiches having an aluminum foam core with the thickness of 15 mm, the skins with three different types of fabrics ([0°/90°] cross ply E-Glass Biaxial stitched, [0°/90°] cross ply E-Glass Woven and [0°/90°] cross ply S-Glass Woven which have same thickness value of 1.75 mm) and two different commercial adhesives (flexible polyurethane and toughened epoxy based) at different values of support span distances (L= 55, 70, 80, 125 mm) by aiming the analyses of their flexural performance. The skins used in the study were produced via Vacuum Assisted Resin Transfer Molding (VARTM) technique and were easily bonded onto the aluminum foam core with flexible and toughened adhesives under a very low pressure using press machine with the alignment tabs having the total thickness of the whole panel. The main results of the flexural loading are: force-displacement curves obtained after the bending tests, peak force values, absorbed energy, collapse mechanisms, adhesion quality and the effect of the support span length and adhesive type. The experimental results presented that the sandwiches with epoxy based toughened adhesive and the skins made of S-Glass Woven fabrics indicated the best adhesion quality and mechanical properties. The sandwiches with toughened adhesive exhibited higher peak force and energy absorption values compared to the sandwiches with flexible adhesive. The core shear mode occurred in the sandwiches with flexible polyurethane based adhesive through the thickness of the core while the same mode took place in the sandwiches with toughened epoxy based adhesive along the length of the core. The use of these sandwich structures can lead to a weight reduction of the transport vehicles, providing an adequate structural strength under operating conditions.

Keywords: adhesive and adhesion, aluminum foam, bending, collapse mechanisms

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540 Physicochemical Properties and Toxicity Studies on a Lectin from the Bulb of Dioscorea bulbifera

Authors: Uchenna Nkiruka Umeononihu, Adenike Kuku, Oludele Odekanyin, Olubunmi Babalola, Femi Agboola, Rapheal Okonji

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In this study, a lectin from the bulb of Dioscorea bulbifera was purified, characterised, and its acute and sub-acute toxicity was investigated with a view to evaluate its toxic effects in mice. The protein from the bulb was extracted by homogenising 50 g of the bulb in 500 ml of phosphate buffered saline (0.025 M) of pH 7.2, stirred for 3 hr, and centrifuged at the speed of 3000 rpm. Blood group and sugar specificity assays of the crude extract were determined. The lectin was purified in a two-step procedure- gel filtration on Sephadex G-75 and affinity chromatography on Sepharose 4-B arabinose. The degree of purity of the purified lectin was ascertained by SDS-polyacrylamide gel electrophoresis. Detection of covalently bound carbohydrate was carried out with Periodic Acid-Schiffs (PAS) reagent staining technique. Effects of temperature, pH, and EDTA on the lectin were carried out using standard methods. This was followed by acute toxicity studies via oral and subcutaneous routes using mice. The animals were monitored for mortality and signs of toxicity. The sub-acute toxicity studies were carried out using rats. Different concentrations of the lectin were administered twice daily for 5 days via the subcutaneous route. The animals were sacrificed on the sixth day; blood samples and liver tissues were collected. Biochemical assays (determination of total protein, direct bilirubin, Alanine aminotransferase (ALT), Aspartate aminotransferase (AST), catalase (CAT), and superoxide dismutase (SOD)) were carried out on the serum and liver homogenates. The collected organs (heart, liver, kidney, and spleen) were subjected to histopathological analysis. The results showed that lectin from the bulbs of Dioscorea bulbifera agglutinated non-specifically the erythrocytes of the human ABO system as well as rabbit erythrocytes. The haemagglutinating activity was strongly inhibited by arabinose and dulcitol with minimum inhibitory concentrations of 0.781 and 6.25, respectively. The lectin was purified to homogeneity with native and subunit molecular weights of 56,273 and 29,373 Daltons, respectively. The lectin was thermostable up to 30 0C and lost 25 %, 33.3 %, and 100 % of its heamagglutinating activity at 40°C, 50°C, and 60°C, respectively. The lectin was maximally active at pH 4 and 5 but lost its total activity at pH eight, while EDTA (10 mM) had no effect on its haemagglutinating activity. PAS reagent staining showed that the lectin was not a glycoprotein. The sub-acute studies on rats showed elevated levels of ALT, AST, serum bilirubin, total protein in serum and liver homogenates suggesting damage to liver and spleen. The study concluded that the aerial bulb of D. bulbifera lectin was non-specific in its heamagglutinating activity and dimeric in its structure. The lectin shared some physicochemical characteristics with lectins from other Dioscorecea species and was moderately toxic to the liver and spleen of treated animals.

Keywords: Dioscorea bulbifera, heamagglutinin, lectin, toxicity

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539 Different Types of Bismuth Selenide Nanostructures for Targeted Applications: Synthesis and Properties

Authors: Jana Andzane, Gunta Kunakova, Margarita Baitimirova, Mikelis Marnauza, Floriana Lombardi, Donats Erts

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Bismuth selenide (Bi₂Se₃) is known as a narrow band gap semiconductor with pronounced thermoelectric (TE) and topological insulator (TI) properties. Unique TI properties offer exciting possibilities for fundamental research as observing the exciton condensate and Majorana fermions, as well as practical application in spintronic and quantum information. In turn, TE properties of this material can be applied for wide range of thermoelectric applications, as well as for broadband photodetectors and near-infrared sensors. Nanostructuring of this material results in improvement of TI properties due to suppression of the bulk conductivity, and enhancement of TE properties because of increased phonon scattering at the nanoscale grains and interfaces. Regarding TE properties, crystallographic growth direction, as well as orientation of the nanostructures relative to the growth substrate, play significant role in improvement of TE performance of nanostructured material. For instance, Bi₂Se₃ layers consisting of randomly oriented nanostructures and/or of combination of them with planar nanostructures show significantly enhanced in comparison with bulk and only planar Bi₂Se₃ nanostructures TE properties. In this work, a catalyst-free vapour-solid deposition technique was applied for controlled obtaining of different types of Bi₂Se₃ nanostructures and continuous nanostructured layers for targeted applications. For example, separated Bi₂Se₃ nanoplates, nanobelts and nanowires can be used for investigations of TI properties; consisting from merged planar and/or randomly oriented nanostructures Bi₂Se₃ layers are useful for applications in heat-to-power conversion devices and infrared detectors. The vapour-solid deposition was carried out using quartz tube furnace (MTI Corp), equipped with an inert gas supply and pressure/temperature control system. Bi₂Se₃ nanostructures/nanostructured layers of desired type were obtained by adjustment of synthesis parameters (process temperature, deposition time, pressure, carrier gas flow) and selection of deposition substrate (glass, quartz, mica, indium-tin-oxide, graphene and carbon nanotubes). Morphology, structure and composition of obtained Bi₂Se₃ nanostructures and nanostructured layers were inspected using SEM, AFM, EDX and HRTEM techniques, as well as home-build experimental setup for thermoelectric measurements. It was found that introducing of temporary carrier gas flow into the process tube during the synthesis and deposition substrate choice significantly influence nanostructures formation mechanism. Electrical, thermoelectric, and topological insulator properties of different types of deposited Bi₂Se₃ nanostructures and nanostructured coatings are characterized as a function of thickness and discussed.

Keywords: bismuth seleinde, nanostructures, topological insulator, vapour-solid deposition

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538 The Influence of Argumentation Strategy on Student’s Web-Based Argumentation in Different Scientific Concepts

Authors: Xinyue Jiao, Yu-Ren Lin

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Argumentation is an essential aspect of scientific thinking which has been widely concerned in recent reform of science education. The purpose of the present studies was to explore the influences of two variables termed ‘the argumentation strategy’ and ‘the kind of science concept’ on student’s web-based argumentation. The first variable was divided into either monological (which refers to individual’s internal discourse and inner chain reasoning) or dialectical (which refers to dialogue interaction between/among people). The other one was also divided into either descriptive (i.e., macro-level concept, such as phenomenon can be observed and tested directly) or theoretical (i.e., micro-level concept which is abstract, and cannot be tested directly in nature). The present study applied the quasi-experimental design in which 138 7th grade students were invited and then assigned to either monological group (N=70) or dialectical group (N=68) randomly. An argumentation learning program called ‘the PWAL’ was developed to improve their scientific argumentation abilities, such as arguing from multiple perspectives and based on scientific evidence. There were two versions of PWAL created. For the individual version, students can propose argument only through knowledge recall and self-reflecting process. On the other hand, the students were allowed to construct arguments through peers’ communication in the collaborative version. The PWAL involved three descriptive science concept-based topics (unit 1, 3 and 5) and three theoretical concept-based topics (unit 2, 4 and 6). Three kinds of scaffoldings were embedded into the PWAL: a) argument template, which was used for constructing evidence-based argument; b) the model of the Toulmin’s TAP, which shows the structure and elements of a sound argument; c) the discussion block, which enabled the students to review what had been proposed during the argumentation. Both quantitative and qualitative data were collected and analyzed. An analytical framework for coding students’ arguments proposed in the PWAL was constructed. The results showed that the argumentation approach has a significant effect on argumentation only in theoretical topics (f(1, 136)=48.2, p < .001, η2=2.62). The post-hoc analysis showed the students in the collaborative group perform significantly better than the students in the individual group (mean difference=2.27). However, there is no significant difference between the two groups regarding their argumentation in descriptive topics. Secondly, the students made significant progress in the PWAL from the earlier descriptive or theoretical topic to the later one. The results enabled us to conclude that the PWAL was effective for students’ argumentation. And the students’ peers’ interaction was essential for students to argue scientifically especially for the theoretical topic. The follow-up qualitative analysis showed student tended to generate arguments through critical dialogue interactions in the theoretical topic which promoted them to use more critiques and to evaluate and co-construct each other’s arguments. More explanations regarding the students’ web-based argumentation and the suggestions for the development of web-based science learning were proposed in our discussions.

Keywords: argumentation, collaborative learning, scientific concepts, web-based learning

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537 Making the Neighbourhood: Analyzing Mapping Procedures to Deal with Plurality and Conflict

Authors: Barbara Roosen, Oswald Devisch

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Spatial projects are often contested. Despite participatory trajectories in official spatial development processes, citizens engage often by their power to say no. Participatory mapping helps to produce more legible and democratic ways of decision-making. It has proven its value in producing a multitude of knowledges and views, for individuals and community groups and local stakeholders to imagine desired and undesired futures and to give them the rhetorical power to present their views throughout the development process. From this perspective, mapping works as a social process in which individuals and groups share their knowledge, learn from each other and negotiate their relationship with each other as well as with space and power. In this way, these processes eventually aim to activate communities to intervene in cooperation in real problems. However, these are fragile and bumpy processes, sometimes leading to (local) conflict and intractable situations. Heterogeneous subjectivities and knowledge that become visible during the mapping process and which are contested by members of the community, is often the first trigger. This paper discusses a participatory mapping project conducted in a residential subdivision in Flanders to provide a deeper understanding of how or under which conditions the mapping process could moderate discordant situations amongst inhabitants, local organisations and local authorities, towards a more constructive outcome. In our opinion, this implies a thorough documentation and presentation of the different steps of the mapping process to design and moderate an open and transparent dialogue. The mapping project ‘Make the Neighbourhood’, is set up in the aftermath of a socio-spatial design intervention in the neighbourhood that led to polarization within the community. To start negotiation between the diverse claims that came to the fore, we co-create a desired future map of the neighbourhood together with local organisations and inhabitants as a way to engage them in the development of a new spatial development plan for the area. This mapping initiative set up a new ‘common’ goal or concern, as a first step to bridge the gap that we experienced between different sociocultural groups, bottom-up and top-down initiatives and between professionals and non-professionals. An atlas of elements (materials), an atlas of actors with different roles and an atlas of ways of cooperation and organisation form the work and building material of the future neighbourhood map, assembled in two co-creation sessions. Firstly, we will consider how the mapping procedures articulate the plurality of claims and agendas. Secondly, we will elaborate upon how social relations and spatialities are negotiated and reproduced during the different steps of the map making. Thirdly, we will reflect on the role of the rules, format, and structure of the mapping process in moderating negotiations between much divided claims. To conclude, we will discuss the challenges of visualizing the different steps of mapping process as a strategy to moderate tense negotiations in a more constructive direction in the context of spatial development processes.

Keywords: conflict, documentation, participatory mapping, residential subdivision

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536 The Decision-Making Mechanisms of Tax Regulations

Authors: Nino Pailodze, Malkhaz Sulashvili, Vladimer Kekenadze, Tea Khutsishvili, Irma Makharashvili, Aleksandre Kekenadze

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In the nearest future among the important problems which Georgia has solve the most important is economic stability, that bases on fiscal policy and the proper definition of the its directions. The main source of the Budget revenue is the national income. The State uses taxes, loans and emission in order to create national income, were the principal weapon are taxes. As well as fiscal function of the fulfillment of the budget, tax systems successfully implement economic and social development and the regulatory functions of foreign economic relations. A tax is a mandatory, unconditional monetary payment to the budget made by a taxpayer in accordance with this Code, based on the necessary, nonequivalent and gratuitous character of the payment. Taxes shall be national and local. National taxes shall be the taxes provided for under this Code, the payment of which is mandatory across the whole territory of Georgia. Local taxes shall be the taxes provided for under this Code, introduced by normative acts of local self-government representative authorities (within marginal rates), the payment of which is mandatory within the territory of the relevant self-governing unit. National taxes have the leading role in tax systems, but also the local taxes have an importance role in tax systems. Exactly in the means of local taxes, the most part of the budget is formatted. National taxes shall be: income tax, profit tax, value added tax (VAT), excise tax, import duty, property tax shall be a local tax The property tax is one of the significant taxes in Georgia. The paper deals with the taxation mechanism that has been operated in Georgia. The above mention has the great influence in financial accounting. While comparing foreign legislation towards Georgian legislation we discuss the opportunity of using their experience. Also, we suggested recommendations in order to improve the tax system in financial accounting. In addition to accounting, which is regulated according the International Accounting Standards we have tax accounting, which is regulated by the Tax Code, various legal orders / regulations of the Minister of Finance. The rules are controlled by the tax authority, Revenue Service. The tax burden from the tax values are directly related to expenditures of the state from the emergence of the first day. Fiscal policy of the state is as well as expenditure of the state and decisions of taxation. In order to get the best and the most effective mobilization of funds, Government’s primary task is to decide the kind of taxation rules. Tax function is to reveal the substance of the act. Taxes have the following functions: distribution or the fiscal function; Control and regulatory functions. Foreign tax systems evolved in the different economic, political and social conditions influence. The tax systems differ greatly from each other: taxes, their structure, typing means, rates, the different levels of fiscal authority, the tax base, the tax sphere of action, the tax breaks.

Keywords: international accounting standards, financial accounting, tax systems, financial obligations

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535 Integrated Approach to Attenuate Insulin Amyloidosis: Synergistic Effects of Peptide and Cysteine Protease Enzymes

Authors: Shilpa Mukundaraj, Nagaraju Shivaiah

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Amyloidogenic conditions, driven by protein aggregation into insoluble fibrils, which pose significant challenges in the clinical condition of diabetes management, particularly through the amyloidogenic LVEALYL sequence in insulin B-chain. This study explores a dual therapeutic strategy involving cysteine protease enzymes such as papain and ficin and inhibitory peptides to target insulin amyloidosis. Combining in silico, in vitro, and in vivo methodologies, the research aims to inhibit amyloid formation and degrade preformed fibrils. Inhibitory peptides were designed using structure-guided approaches in Rosetta to specifically target the LVEALYL sequence. Concurrently, cysteine protease enzymes, including papain and ficin, were evaluated for their fibril disassembly potential. In vitro experiments, utilizing SDS- PAGE and spectroscopic techniques, confirmed dose-dependent degradation 50 to 300ug in vitro and 60mg/kg in vivo of amyloid aggregates by these enzymes, with significant disaggregation observed at higher concentrations 20mg. Peptide inhibitors effectively reduced fibril formation, as evidenced by reduced Thioflavin T fluorescence and circular dichroism spectroscopy. Complementary in silico analyses, including molecular docking and dynamic simulations, provided structural insights into enzyme binding interactions with amyloidogenic regions. Key residues involved in substrate recognition and cleavage were identified, with computational findings aligning strongly with experimental data. These insights confirmed the specificity of papain and ficin in targeting insulin fibrils. For translational potential, an in vivo rat model was developed, involving subcutaneous insulin amyloid injections to induce localized amyloid deposits. Over six days of enzyme treatment, a marked reduction in amyloid burden was observed through histological findings and biochemical assay superoxide dismutase can provide insights into oxidative damage due to amyloid deposition. Furthermore, inflammatory markers IL-6, TNFα were significantly attenuated in treated groups, emphasizing the dual role of enzymes in amyloid clearance and inflammation modulation. This integrative study highlights the promise of cysteine protease enzymes and inhibitory peptides as complementary therapeutic strategies for managing insulin amyloidosis. By targeting both the formation and persistence of amyloid fibrils, this dual approach offers a novel and effective avenue for amyloidosis treatment.

Keywords: insulin amyloidosis, peptide inhibitors, cysteine protease enzymes, amyloid degradation

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534 The Mitigation of Quercetin on Lead-Induced Neuroinflammation in a Rat Model: Changes in Neuroinflammatory Markers and Memory

Authors: Iliyasu Musa Omoyine, Musa Sunday Abraham, Oladele Sunday Blessing, Iliya Ibrahim Abdullahi, Ibegbu Augustine Oseloka, Nuhu Nana-Hawau, Animoku Abdulrazaq Amoto, Yusuf Abdullateef Onoruoiza, Sambo Sohnap James, Akpulu Steven Peter, Ajayi Abayomi

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The neuroprotective role of inflammation from detrimental intrinsic and extrinsic factors has been reported. However, the overactivation of astrocytes and microglia due to lead toxicity produce excessive pro-inflammatory cytokines, mediating neurodegenerative diseases. The present study investigated the mitigatory effects of quercetin on neuroinflammation, correlating with memory function in lead-exposed rats. In this study, Wistar rats were administered orally with Quercetin (Q: 60 mg/kg) and Succimer as a standard drug (S: 10 mg/kg) for 21 days after lead exposure (Pb: 125 mg/kg) of 21 days or in combination with Pb, once daily for 42 days. Working and reference memory was assessed using an Eight-arm radial water maze (8-ARWM). The changes in brain lead level, the neuronal nitric oxide synthase (nNOS) activity, and the level of neuroinflammatory markers such as tumour necrosis factor-alpha (TNF-α) and Interleukin 1 Beta (IL-1β) were determined. Immunohistochemically, astrocyte expression was evaluated. The results showed that the brain level of lead was increased significantly in lead-exposed rats. The expression of astrocytes increased in the CA3 and CA1 regions of the hippocampus, and the levels of brain TNF-α and IL-1β increased in lead-exposed rats. Lead impaired reference and working memory by increasing reference memory errors and working memory incorrect errors in lead-exposed rats. However, quercetin treatment effectively improved memory and inhibited neuroinflammation by reducing astrocytes’ expression and the levels of TNF-α and IL-1β. The expression of astrocytes and the levels of TNF-α and IL-1β correlated with memory function. The possible explanation for quercetin’s anti-neuroinflammatory effect is that it modulates the activity of cellular proteins involved in the inflammatory response; inhibits the transcription factor of nuclear factor-kappa B (NF-κB), which regulates the expression of proinflammatory molecules; inhibits kinases required for the synthesis of Glial fibrillary acidic protein (GFAP) and modifies the phosphorylation of some proteins, which affect the structure and function of intermediate filament proteins; and, lastly, induces Cyclic-AMP Response Element Binding (CREB) activation and neurogenesis as a compensatory mechanism for memory deficits and neuronal cell death. In conclusion, the levels of neuroinflammatory markers negatively correlated with memory function. Thus, quercetin may be a promising therapy in neuroinflammation and memory dysfunction in populations prone to lead exposure.

Keywords: lead, quercetin, neuroinflammation, memory

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533 A Computational Approach to Screen Antagonist’s Molecule against Mycobacterium tuberculosis Lipoprotein LprG (Rv1411c)

Authors: Syed Asif Hassan, Tabrej Khan

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Tuberculosis (TB) caused by bacillus Mycobacterium tuberculosis (Mtb) continues to take a disturbing toll on human life and healthcare facility worldwide. The global burden of TB remains enormous. The alarming rise of multi-drug resistant strains of Mycobacterium tuberculosis calls for an increase in research efforts towards the development of new target specific therapeutics against diverse strains of M. tuberculosis. Therefore, the discovery of new molecular scaffolds targeting new drug sites should be a priority for a workable plan for fighting resistance in Mycobacterium tuberculosis (Mtb). Mtb non-acylated lipoprotein LprG (Rv1411c) has a Toll-like receptor 2 (TLR2) agonist actions that depend on its association with triacylated glycolipids binding specifically with the hydrophobic pocket of Mtb LprG lipoprotein. The detection of a glycolipid carrier function has important implications for the role of LprG in Mycobacterial physiology and virulence. Therefore, considering the pivotal role of glycolipids in mycobacterial physiology and host-pathogen interactions, designing competitive antagonist (chemotherapeutics) ligands that competitively bind to glycolipid binding domain in LprG lipoprotein, will lead to inhibition of tuberculosis infection in humans. In this study, a unified approach involving ligand-based virtual screening protocol USRCAT (Ultra Shape Recognition) software and molecular docking studies using Auto Dock Vina 1.1.2 using the X-ray crystal structure of Mtb LprG protein was implemented. The docking results were further confirmed by DSX (DrugScore eXtented), a robust program to evaluate the binding energy of ligands bound to the Ligand binding domain of the Mtb LprG lipoprotein. The ligand, which has the higher hypothetical affinity, also has greater negative value. Based on the USRCAT, Lipinski’s values and molecular docking results, [(2R)-2,3-di(hexadecanoyl oxy)propyl][(2S,3S,5S,6R)-3,4,5-trihydroxy-2,6-bis[[(2R,3S,4S,5R,6S)-3,4,5-trihydroxy-6 (hydroxymethyl)tetrahydropyran-2-yl]oxy]cyclohexyl] phosphate (XPX) was confirmed as a promising drug-like lead compound (antagonist) binding specifically to the hydrophobic domain of LprG protein with affinity greater than that of PIM2 (agonist of LprG protein) with a free binding energy of -9.98e+006 Kcal/mol and binding affinity of -132 Kcal/mol, respectively. A further, in vitro assay of this compound is required to establish its potency in inhibiting molecular evasion mechanism of MTB within the infected host macrophages. These results will certainly be helpful in future anti-TB drug discovery efforts against Multidrug-Resistance Tuberculosis (MDR-TB).

Keywords: antagonist, agonist, binding affinity, chemotherapeutics, drug-like, multi drug resistance tuberculosis (MDR-TB), RV1411c protein, toll-like receptor (TLR2)

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532 The Political Economy of the Global Climate Change Adaptation Initiatives: A Case Study on the Global Environmental Facility

Authors: Anar Koli

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After the Paris agreement in 2015, a comprehensive initiative both from the developed and developing countries towards the adaptation to climate change is emerging. The Global Environmental Facility (GEF), which is financing a global portfolio of adaptation projects and programs in over 124 countries is playing a significant role to a new financing framework that included the concept of “climate-resilient development”. However, both the adaptation and sustainable development paradigms remain continuously contested, especially the role of the multilateral institutions with their technical and financial assistance to the developing world. Focusing on the adaptation initiatives of the GEF, this study aims to understand to what extent the global multilateral institutions, particularly the GEF is contributing to the climate-resilient development. From the political ecology perspective, the argument of this study is that the global financial framework is highly politicized, and understanding the contribution of the global institutions of the global climate change needs to be related both from the response and causal perspectives. A holistic perspective, which includes the contribution of the GEF as a response to the climate change and as well the cause of global climate change, are needed to understand the broader environment- political economic relation. The study intends to make a critical analysis of the way in which the political economy structure and the environment are related along with the social and ecological implications. It does not provide a narrow description of institutional responses to climate change, rather it looks at how the global institutions are influencing the relationship of the global ecologies and economies. This study thus developed a framework combining the global governance and the political economy perspective. This framework includes environment-society relation, environment-political economy linkage, global institutions as the orchestra, and division between the North and the South. Through the analysis of the GEF as the orchestra of the global governance, this study helps to understand how GEF is coordinating the interactions between the North and the South and responding the global climate resilient development. Through the other components of the framework, the study explains how the role of the global institutions is related to the cause of the human induced global climate change. The study employs a case study based on both the quantitative and qualitative data. Along with the GEF reports and data sets, this study draws from an eclectic range of literature from a range of disciplines to explain the broader relation of the environment and political economy. Based on a case study on GEF, the study found that the GEF has positive contributions in bringing developing countries’ capacity in terms of sustainable development goal, local institutional development. However, through a critical holistic analysis, this study found that this contribution to the resilient development helps the developing countries to conform the fossil fuel based capitalist political economy. The global governance institution is contributing both to the pro market based environment society relation and, to the consequences of this relation.

Keywords: climate change adaptation, global environmental facility (GEF), political economy, the north -south relation

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531 Investigating the Relationship between Job Satisfaction, Role Identity, and Turnover Intention for Nurses in Outpatient Department

Authors: Su Hui Tsai, Weir Sen Lin, Rhay Hung Weng

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There are numerous outpatient departments at hospitals with enormous amounts of outpatients. Although the work of outpatient nursing staff does not include the ward, emergency and critical care units that involve patient life-threatening conditions, the work is cumbersome and requires facing and dealing with a large number of outpatients in a short period of time. Therefore, nursing staff often do not feel satisfied with their work and cannot identify with their professional role, leading to intentions to leave their job. Thus, the main purpose of this study is to explore the correlation between the job satisfaction and role identity of nursing staff with turnover intention. This research was conducted using a questionnaire, and the subjects were outpatient nursing staff in three regional hospitals in Southern Taiwan. A total of 175 questionnaires were distributed, and 166 valid questionnaires were returned. After collecting the data, the reliability and validity of the study variables were confirmed by confirmatory factor analysis. The influence of role identity and job satisfaction on nursing staff’s turnover intention was analyzed by descriptive analysis, one-way ANOVA, Pearson correlation analysis and multiple regression analysis. Results showed that 'role identity' had significant differences in different types of marriages. Job satisfaction of 'grasp of environment' had significant differences in different levels of education. Job satisfaction of 'professional growth' and 'shifts and days off' showed significant differences in different types of marriages. 'Role identity' and 'job satisfaction' were negatively correlated with turnover intention respectively. Job satisfaction of 'salary and benefits' and 'grasp of environment' were significant predictors of role identity. The higher the job satisfaction of 'salary and benefits' and 'grasp of environment', the higher the role identity. Job satisfaction of 'patient and family interaction' were significant predictors of turnover intention. The lower the job satisfaction of 'patient and family interaction', the higher the turnover intention. This study found that outpatient nursing staff had the lowest satisfaction towards salary structure. It is recommended that bonuses, promotion opportunities and other incentives be established to increase the role identity of outpatient nursing staff. The results showed that the higher the job satisfaction of 'salary and benefits' and 'grasp of environment', the higher the role identity. It is recommended that regular evaluations be conducted to reward nursing staff with excellent service and invite nursing staff to share their work experiences and thoughts, to enhance nursing staff’s expectation and identification of their occupational role, as well as instilling the concept of organizational service and organizational expectations of emotional display. The results showed that the lower the job satisfaction of 'patient and family interaction', the higher the turnover intention. It is recommended that interpersonal communication and workplace violence prevention educational training courses be organized to enhance the communication and interaction of nursing staff with patients and their families.

Keywords: outpatient, job satisfaction, turnover, intention

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530 Financial Analysis of the Foreign Direct in Mexico

Authors: Juan Peña Aguilar, Lilia Villasana, Rodrigo Valencia, Alberto Pastrana, Martin Vivanco, Juan Peña C

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Each year a growing number of companies entering Mexico in search of the domestic market share. These activities, including stores, telephone long distance and local raw materials and energy, and particularly the financial sector, have managed to significantly increase its weight in the flows of FDI in Mexico , however, you should consider whether these trends FDI are positive for the Mexican economy and these activities increase Mexican exports in the medium term , and its share in GDP , gross fixed capital formation and employment. In general stresses that these activities, by far, have been unable to significantly generate linkages with the rest of the economy, a process that has not favored with competitiveness policies and activities aimed at these neutral or horizontal. Since the nineties foreign direct investment (FDI) has shown a remarkable dynamism, both internationally and in Latin America and in Mexico. Only in Mexico the first recipient of FDI in importance in Latin America during 1990-1995 and was displaced by Brazil since FDI increased from levels below 1 % of GDP during the eighties to around 3 % of GDP during the nineties. Its impact has been significant not only from a macroeconomic perspective , it has also allowed the generation of a new industrial production structure and organization, parallel to a significant modernization of a segment of the economy. The case of Mexico also is particularly interesting and relevant because the destination of FDI until 1993 had focused on the purchase of state assets during privatization process. This paper aims to present FDI flows in Mexico and analyze the different business strategies that have been touched and encouraged by the FDI. On the one hand, looking briefly discuss regulatory issues and source and recipient of FDI sectors. Furthermore, the paper presents in more detail the impacts and changes that generated the FDI contribution of FDI in the Mexican economy , besides the macroeconomic context and later legislative changes that resulted in the current regulations is examined around FDI in Mexico, including aspects of the Free Trade Agreement (NAFTA). It is worth noting that foreign investment can not only be considered from the perspective of the receiving economic units. Instead, these flows also reflect the strategic interests of transnational corporations (TNCs) and other companies seeking access to markets and increased competitiveness of their production networks and global distribution, among other reasons. Similarly it is important to note that foreign investment in its various forms is critically dependent on historical and temporal aspects. Thus, the same functionality can vary significantly depending on the specific characteristics of both receptor units as sources of FDI, including macroeconomic, institutional, industrial organization, and social aspects, among others.

Keywords: foreign direct investment (FDI), competitiveness, neoliberal regime, globalization, gross domestic product (GDP), NAFTA, macroeconomic

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529 Immobilization of β-Galactosidase from Kluyveromyces Lactis on Polyethylenimine-Agarose for Production of Lactulose

Authors: Carlos A. C. G. Neto, Natan C. G. Silva, Thais O. Costa, Luciana R. B. Goncalves, Maria v. P. Rocha

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Galactosidases are enzymes responsible for catalyzing lactose hydrolysis reactions and also favoring transgalactosylation reactions for the production of prebiotics, among which lactulose stands out. These enzymes, when immobilized, can have some enzymatic characteristics substantially improved, and the coating of supports with multifunctional polymers in immobilization processes is a promising alternative in order to extend the useful life of the biocatalysts, for example, the coating with polyethyleneimine (PEI). PEI is a flexible polymer that suits the structure of the enzyme, giving greater stability, especially for multimeric enzymes such as β-galactosidases and also protects it from environmental variations, for example, pH and temperature. In addition, it can substantially improve the immobilization parameters and also the efficiency of enzymatic reactions. In this context, the aim of the present work was first to develop biocatalysts of β-galactosidase from Kluyveromyces lactis immobilized on PEI coated agarose, determining the immobilization parameters, its operational and thermal stability, and then to apply it in the hydrolysis of lactose and synthesis of lactulose, using whey as a substrate. This immobilization strategy was chosen in order to improve the catalytic efficiency of the enzyme in the transgalactosylation reaction for the production of prebiotics, and there are few studies with β-galactosidase from this strain. The immobilization of β-galactosidase in agarose previously functionalized with 48% (w/v) glycidol and then coated with 10% (w/v) PEI solution was evaluated using an enzymatic load of 10 mg/g of protein. Subsequently, the hydrolysis and transgalactosylation reactions were conducted at 50 °C, 120 RPM for 20 minutes, using whey (66.7 g/L of lactose) supplemented with 133.3 g/L fructose at a ratio of 1:2 (lactose/fructose). Operational stability studies were performed in the same conditions for 10 cycles. Thermal stabilities of biocatalysts were conducted at 50 ºC in 50 mM phosphate buffer, pH 6.6, with 0.1 mM MnCl2. The biocatalysts whose supports were coated were named AGA_GLY_PEI_GAL, and those that were not coated were named AGA_GLY_GAL. The coating of the support with PEI considerably improved immobilization yield (2.6-fold), the biocatalyst activity (1.4-fold), and efficiency (2.2-fold). The biocatalyst AGA_GLY_PEI_GAL was better than AGA_GLY_GAL in hydrolysis and transgalactosylation reactions, converting 88.92% of lactose at 5 min of reaction and obtaining a residual concentration of 5.24 g/L. Besides that, it was produced 13.90 g/L lactulose in the same time interval. AGA_GLY_PEI_GAL biocatalyst was stable during the 10 cycles evaluated, converting approximately 80% of lactose and producing 10.95 g/L of lactulose even after the tenth cycle. However, the thermal stability of AGA_GLY_GAL biocatalyst was superior, with a half-life time 5 times higher, probably because the enzyme was immobilized by covalent bonding, which is stronger than adsorption (AGA_GLY_PEI_GAL). Therefore, the strategy of coating the supports with PEI has proven to be effective for the immobilization of β-galactosidase from K. lactis, considerably improving the immobilization parameters, as well as the enzyme, catalyzed reactions. In addition, the use of whey as a raw material for lactulose production has proved to be an industrially advantageous alternative.

Keywords: β-galactosidase, immobilization, lactulose, polyethylenimine, whey

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528 Melt–Electrospun Polyprophylene Fabrics Functionalized with TiO2 Nanoparticles for Effective Photocatalytic Decolorization

Authors: Z. Karahaliloğlu, C. Hacker, M. Demirbilek, G. Seide, E. B. Denkbaş, T. Gries

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Currently, textile industry has played an important role in world’s economy, especially in developing countries. Dyes and pigments used in textile industry are significant pollutants. Most of theirs are azo dyes that have chromophore (-N=N-) in their structure. There are many methods for removal of the dyes from wastewater such as chemical coagulation, flocculation, precipitation and ozonation. But these methods have numerous disadvantages and alternative methods are needed for wastewater decolorization. Titanium-mediated photodegradation has been used generally due to non-toxic, insoluble, inexpensive, and highly reactive properties of titanium dioxide semiconductor (TiO2). Melt electrospinning is an attractive manufacturing process for thin fiber production through electrospinning from PP (Polyprophylene). PP fibers have been widely used in the filtration due to theirs unique properties such as hydrophobicity, good mechanical strength, chemical resistance and low-cost production. In this study, we aimed to investigate the effect of titanium nanoparticle localization and amine modification on the dye degradation. The applicability of the prepared chemical activated composite and pristine fabrics for a novel treatment of dyeing wastewater were evaluated.In this study, a photocatalyzer material was prepared from nTi (titanium dioxide nanoparticles) and PP by a melt-electrospinning technique. The electrospinning parameters of pristine PP and PP/nTi nanocomposite fabrics were optimized. Before functionalization with nTi, the surface of fabrics was activated by a technique using glutaraldehyde (GA) and polyethyleneimine to promote the dye degredation. Pristine PP and PP/nTi nanocomposite melt-electrospun fabrics were characterized using scanning electron microscopy (SEM) and X-Ray Photon Spectroscopy (XPS). Methyl orange (MO) was used as a model compound for the decolorization experiments. Photocatalytic performance of nTi-loaded pristine and nanocomposite melt-electrospun filters was investigated by varying initial dye concentration 10, 20, 40 mg/L). nTi-PP composite fabrics were successfully processed into a uniform, fibrous network of beadless fibers with diameters of 800±0.4 nm. The process parameters were determined as a voltage of 30 kV, a working distance of 5 cm, a temperature of the thermocouple and hotcoil of 260–300 ºC and a flow rate of 0.07 mL/h. SEM results indicated that TiO2 nanoparticles were deposited uniformly on the nanofibers and XPS results confirmed the presence of titanium nanoparticles and generation of amine groups after modification. According to photocatalytic decolarization test results, nTi-loaded GA-treated pristine or nTi-PP nanocomposite fabric filtern have superior properties, especially over 90% decolorization efficiency at GA-treated pristine and nTi-PP composite PP fabrics. In this work, as a photocatalyzer for wastewater treatment, surface functionalized with nTi melt-electrospun fabrics from PP were prepared. Results showed melt-electrospun nTi-loaded GA-tretaed composite or pristine PP fabrics have a great potential for use as a photocatalytic filter to decolorization of wastewater and thus, requires further investigation.

Keywords: titanium oxide nanoparticles, polyprophylene, melt-electrospinning

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527 Psychogeographic Analysis of Campus Design: Spatial Appropriation via Walking Practice in the Cases of Van Yüzüncü Yıl University and Ankara Middle East Technical University in Turkey

Authors: Yasemin İlkay

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Street is not only a crucial spatial unit in urban design and planning discipline but also the context of walking practice in urban space. Moreover, psychogeography concentrates on both ‘walking’ and, therefore, the differentiated forms of (urban) streets to examine the influence of the built environment on the feelings and attitudes of human beings. This paper focuses on ‘walking practice’ in university campuses with reference to spatial appropriation forms via a psychogeographic lens on the phenomenon of alle in two different cities of Turkey, Ankara, the capital city, and Van, in the eastern part of the country. Alle, as an extension of ‘street’ in university campuses, is the constructive spatial structure in university campuses, and as a result, it should be the (both physical and mental) spine of design policy while conceiving and constructing a university campus. The main question of the paper is: How does the interrelation of ‘campus design’ and ‘walking practice’ on alle penetrate reciprocally on the spatial representations of citizens within their urban daily lives. The body contacts with and at urban space (with other objects and subjects) via its movements and stops; this interaction occurs through the spatial pattern of occupancy and vacancy. Walking practice leads to a set of cognitive mental representations in relation to the repertoire of place attachment and spatial appropriation. University campuses are autonomous and fruitful urban spaces to investigate such an interaction. There are both physical/real and psychogeographic representations of the same urban spaces and urban spatial practices. This separation would indicate the invisible dimensions of the difference between ‘what is conceived’ and ‘what is perceived.’ This study aims to compare and contrast the role of alle in both campus design and spatial appropriation via walking at two differentiated university campuses by collecting the mental representations, doing in-depth interviews, and attending walks with the interviewees by psychogeographic techniques. Campus design and spatial appropriation will be compared [with reference to the conception and perception of alle] in three scales: (1) the historical spatial development stories and design approaches of university campuses, (2) the spatial pattern of campuses on the basis of alle, and (3) sub-behavioral regions of the alle in campuses in relation with mental representations and psychogeographic attentive walks. The sub-questions of the research are: [1] How and why do the design approaches differentiate in two university campuses in Turkey, [2] How the interrelation among alle design and spatial appropriation differs in these two cases, and [3] What do the differentiated gaps among real and psychographic maps indicate about the design and spatial appropriation interrelation. METU, as a well-designed, readable campus with its alle, promise a rich walking practice with in-depth and fruitful spatial appropriation regions; however, Van YYÜ limits both the practice and place attachment with its partial design with an alle which is later added to the campus. This research both displays the role of alle in the campus design, walking practice and spatial appropriation and opens a new methodological path to discover hidden knowledge within urban spaces.

Keywords: alle, campus design, cognitive geography, psychogeography, spatial appropriation, Turkey

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526 Influence of Structured Capillary-Porous Coatings on Cryogenic Quenching Efficiency

Authors: Irina P. Starodubtseva, Aleksandr N. Pavlenko

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Quenching is a term generally accepted for the process of rapid cooling of a solid that is overheated above the thermodynamic limit of the liquid superheat. The main objective of many previous studies on quenching is to find a way to reduce the total time of the transient process. Computational experiments were performed to simulate quenching by a falling liquid nitrogen film of an extremely overheated vertical copper plate with a structured capillary-porous coating. The coating was produced by directed plasma spraying. Due to the complexities in physical pattern of quenching from chaotic processes to phase transition, the mechanism of heat transfer during quenching is still not sufficiently understood. To our best knowledge, no information exists on when and how the first stable liquid-solid contact occurs and how the local contact area begins to expand. Here we have more models and hypotheses than authentically established facts. The peculiarities of the quench front dynamics and heat transfer in the transient process are studied. The created numerical model determines the quench front velocity and the temperature fields in the heater, varying in space and time. The dynamic pattern of the running quench front obtained numerically satisfactorily correlates with the pattern observed in experiments. Capillary-porous coatings with straight and reverse orientation of crests are investigated. The results show that the cooling rate is influenced by thermal properties of the coating as well as the structure and geometry of the protrusions. The presence of capillary-porous coating significantly affects the dynamics of quenching and reduces the total quenching time more than threefold. This effect is due to the fact that the initialization of a quench front on a plate with a capillary-porous coating occurs at a temperature significantly higher than the thermodynamic limit of the liquid superheat, when a stable solid-liquid contact is thermodynamically impossible. Waves present on the liquid-vapor interface and protrusions on the complex micro-structured surface cause destabilization of the vapor film and the appearance of local liquid-solid micro-contacts even though the average integral surface temperature is much higher than the liquid superheat limit. The reliability of the results is confirmed by direct comparison with experimental data on the quench front velocity, the quench front geometry, and the surface temperature change over time. Knowledge of the quench front velocity and total time of transition process is required for solving practically important problems of nuclear reactors safety.

Keywords: capillary-porous coating, heat transfer, Leidenfrost phenomenon, numerical simulation, quenching

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525 Cloning and Expression a Gene of β-Glucosidase from Penicillium echinulatum in Pichia pastoris

Authors: Amanda Gregorim Fernandes, Lorena Cardoso Cintra, Rosalia Santos Amorim Jesuino, Fabricia Paula De Faria, Marcio José Poças Fonseca

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Bioethanol is one of the most promising biofuels and able to replace fossil fuels and reduce its different environmental impacts and can be generated from various agroindustrial waste. The Brazil is in first place in bioethanol production to be the largest producer of sugarcane. The bagasse sugarcane (SCB) has lignocellulose which is composed of three major components: cellulose, hemicellulose and lignin. Cellulose is a homopolymer of glucose units connected by glycosidic linkages. Among all species of Penicillium, Penicillium echinulatum has been the focus of attention because they produce high quantities of cellulase and the mutant strain 9A02S1 produces higher enzyme levels compared to the wild. Among the cellulases, the cellobiohydrolases enzymes are the main components of the cellulolytic system of fungi, and are also responsible for most of the potential hydrolytic in enzyme cocktails for the industrial processing of plant biomass and several cellobiohydrolases Penicillium had higher specific activity against cellulose compared to CBH I from Trichoderma reesei. This fact makes it an interesting pattern for higher yields in the enzymatic hydrolysis, and also they are important enzymes in the hydrolysis of crystalline regions of cellulose. Therefore, finding new and more active enzymes become necessary. Meanwhile, β-glycosidases act on soluble substrates and are highly dependent on cellobiohydrolases and endoglucanases action to provide the substrate in the hydrolysis of the biomass, but the cellobiohydrolases and endoglucanases are highly dependent β-glucosidases to maintain efficient hydrolysis. Thus, there is a need to understand the structure-function relationships that govern the catalytic activity of cellulolytic enzymes to elucidate its mechanism of action and optimize its potential as industrial biocatalysts. To evaluate the enzyme β-glucosidase of Penicillium echinulatum (PeBGL1) the gene was synthesized from the assembly sequence from a library in induction conditions and then the PeBGL1 gene was cloned in the vector pPICZαA and transformed into P. pastoris GS115. After processing, the producers of PeBGL1 were analyzed for enzyme activity and protein profile where a band of approximately 100 kDa was viewed. It was also carried out the zymogram. In partial characterization it was determined optimum temperature of 50°C and optimum pH of 6,5. In addition, to increase the secreted recombinant PeBGL1 production by Pichia pastoris, three parameters of P. pastoris culture medium were analysed: methanol, nitrogen source concentrations and the inoculum size. A 23 factorial design was effective in achieving the optimum condition. Altogether, these results point to the potential application of this P. echinulatum β-glucosidase in hydrolysis of cellulose for the production of bioethanol.

Keywords: bioethanol, biotechnology, beta-glucosidase, penicillium echinulatum

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524 Fructose-Aided Cross-Linked Enzyme Aggregates of Laccase: An Insight on Its Chemical and Physical Properties

Authors: Bipasa Dey, Varsha Panwar, Tanmay Dutta

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Laccase, a multicopper oxidase (EC 1.10.3.2) have been at the forefront as a superior industrial biocatalyst. They are versatile in terms of bestowing sustainable and ecological catalytic reactions such as polymerisation, xenobiotic degradation and bioremediation of phenolic and non-phenolic compounds. Regardless of the wide biotechnological applications, the critical limiting factors viz. reusability, retrieval, and storage stability still prevail. This can cause an impediment in their applicability. Crosslinked enzyme aggregates (CLEAs) have emerged as a promising technique that rehabilitates these essential facets, albeit at the expense of their enzymatic activity. The carrier free crosslinking method prevails over the carrier-bound immobilisation in conferring high productivity, low production cost owing to the absence of additional carrier and circumvent any non-catalytic ballast which could dilute the volumetric activity. To the best of our knowledge, the ε-amino group of lysyl residue is speculated as the best choice for forming Schiff’s base with glutaraldehyde. Despite being most preferrable, excess glutaraldehyde can bring about disproportionate and undesirable crosslinking within the catalytic site and hence could deliver undesirable catalytic losses. Moreover, the surface distribution of lysine residues in Trametes versicolor laccase is significantly less. Thus, to mitigate the adverse effect of glutaraldehyde in conjunction with scaling down the degradation or catalytic loss of the enzyme, crosslinking with inert substances like gelatine, collagen, Bovine serum albumin (BSA) or excess lysine is practiced. Analogous to these molecules, sugars have been well known as a protein stabiliser. It helps to retain the structural integrity, specifically secondary structure of the protein during aggregation by changing the solvent properties. They are comprehended to avert protein denaturation or enzyme deactivation during precipitation. We prepared crosslinked enzyme aggregates (CLEAs) of laccase from T. versicolor with the aid of sugars. The sugar CLEAs were compared with the classic BSA and glutaraldehyde laccase CLEAs concerning physico-chemical properties. The activity recovery for the fructose CLEAs were found to be ~20% higher than the non-sugar CLEA. Moreover, the 𝐾𝑐𝑎𝑡𝐾𝑚⁄ values of the CLEAs were two and three-fold higher than BSA-CLEA and GACLEA, respectively. The half-life (t1/2) deciphered by sugar-CLEA was higher than the t1/2 of GA-CLEAs and free enzyme, portraying more thermal stability. Besides, it demonstrated extraordinarily high pH stability, which was analogous to BSA-CLEA. The promising attributes of increased storage stability and recyclability (>80%) gives more edge to the sugar-CLEAs over conventional CLEAs of their corresponding free enzyme. Thus, sugar-CLEA prevails in furnishing the rudimentary properties required for a biocatalyst and holds many prospects.

Keywords: cross-linked enzyme aggregates, laccase immobilization, enzyme reusability, enzyme stability

Procedia PDF Downloads 103