Search results for: buffer overflow

Authors: Tesfay Gidey Bezabeh, Tânia Sofia Oliveira, Josep Crous-Duran, João H. N. Palma

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Ethiopia's economy depends strongly on Coffea arabica production. Coffee, like many other crops, is sensitive to climate change. An urgent development and application of strategies against the negative impacts of climate change on coffee production is important. Agroforestry-based system is one of the strategies that may ensure sustainable coffee production amidst the likelihood of future impacts of climate change. This system involves the combination of trees in buffer extremes, thereby modifying microclimate conditions. This paper assessed coffee production under 1) coffee monoculture and 2) coffee grown using an agroforestry system, under a) current climate and b) two different future climate change scenarios. The study focused on two representative coffee-growing regions of Ethiopia under different soil, climate, and elevation conditions. A process-based growth model (Yield-SAFE) was used to simulate coffee production for a time horizon of 40 years. Climate change scenarios considered were representative concentration pathways (RCP) 4.5 and 8.5. The results revealed that in monoculture systems, the current coffee yields are between 1200-1250 kg ha⁻¹ yr⁻¹, with an expected decrease between 4-38% and 20-60% in scenarios RCP 4.5 and 8.5, respectively. However, in agroforestry systems, the current yields are between 1600-2200 kg ha⁻¹ yr⁻¹; the decrease was lower, ranging between 4-13% and 16-25% in RCP 4.5 and 8.5 scenarios, respectively. From the results, it can be concluded that coffee production under agroforestry systems has a higher level of resilience when facing future climate change and reinforces the idea of using this type of management in the near future for adapting climate change's negative impacts on coffee production.

Keywords: Albizia gummifera, CORDEX, Ethiopia, HADCM3 model, process-based model

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Authors: Endang Lukitaningsih, Fathul Jannah, Arief R. Hakim, Ratna D. Puspita, Zullies Ikawati

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Rifampicin is a semisynthetic antibiotic derivative of rifamycin B produced by Streptomyces mediterranei. RIF has been used worldwide as first line drug-prescribed throughout tuberculosis therapy. This study aims to develop and to validate an HPLC method couple with a UV detection for determination of rifampicin in spiked human plasma and its application for bioequivalence study. The chromatographic separation was achieved on an RP-C18 column (LachromHitachi, 250 x 4.6 mm., 5μm), utilizing a mobile phase of phosphate buffer/acetonitrile (55:45, v/v, pH 6.8 ± 0.1) at a flow of 1.5 mL/min. Detection was carried out at 337 nm by using spectrophotometer. The developed method was statistically validated for the linearity, accuracy, limit of detection, limit of quantitation, precise and specifity. The specifity of the method was ascertained by comparing chromatograms of blank plasma and plasma containing rifampicin; the matrix and rifampicin were well separated. The limit of detection and limit of quantification were 0.7 µg/mL and 2.3 µg/mL, respectively. The regression curve of standard was linear (r > 0.999) over a range concentration of 20.0 – 100.0 µg/mL. The mean recovery of the method was 96.68 ± 8.06 %. Both intraday and interday precision data showed reproducibility (R.S.D. 2.98% and 1.13 %, respectively). Therefore, the method can be used for routine analysis of rifampicin in human plasma and in bioequivalence study. The validated method was successfully applied in pharmacokinetic and bioequivalence study of rifampicin tablet in a limited number of subjects (under an Ethical Clearance No. KE/FK/6201/EC/2015). The mean values of Cmax, Tmax, AUC(0-24) and AUC(o-∞) for the test formulation of rifampicin were 5.81 ± 0.88 µg/mL, 1.25 hour, 29.16 ± 4.05 µg/mL. h. and 29.41 ± 4.07 µg/mL. h., respectively. Meanwhile for the reference formulation, the values were 5.04 ± 0.54 µg/mL, 1.31 hour, 27.20 ± 3.98 µg/mL.h. and 27.49 ± 4.01 µg/mL.h. From bioequivalence study, the 90% CIs for the test formulation/reference formulation ratio for the logarithmic transformations of Cmax and AUC(0-24) were 97.96-129.48% and 99.13-120.02%, respectively. According to the bioequivamence test guidelines of the European Commission-European Medicines Agency, it can be concluded that the test formulation of rifampicin is bioequivalence with the reference formulation.

Keywords: validation, HPLC, plasma, bioequivalence

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Authors: Asma Batool

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Immigrant families raising a child with developmental disabilities in Canada encounter many challenges during the process of disability care. Starting from the early screening of their child for diagnosis followed by challenges associated with treatment, access and service utilization. A substantial amount of research focuses on identifying barriers. However, the functional aspects of barriers in terms of their potential influences on parents and children with disabilities are unexplored yet. This paper presents functional analysis of barriers in disability care research by adopting a method of integrated approach. Juxtaposition of two developmental approaches, Bronfenbrenner’s ecological model and parents ‘transformational process model is generating multiple hypotheses to be considered while empirically investigating causal relationships and mediating or moderating factors among various variables related with disability care research. This functional analysis suggests that barriers have negative impacts on the physical and emotional development of children with disabilities as well as on the overall quality of family life (QOFL). While, barriers have facilitating impacts on parents, alternatively, the process of transformation in parents expedite after experiencing barriers. Consequently, parents reconstruct their philosophy of life and experience irreversible but continuous developmental change in terms of transformations simultaneously with their developing child and may buffer the expected negative impacts of barriers on disabled child and QOFL. Overall, this paper is suggesting implications for future research and parents’ transformations are suggesting potential pathways to minimize the negative influences of barriers that parents experience during disability care, hence improving satisfaction in QOFL in general.

Keywords: barriers in disability care, developmental disabilities, parents’ transformations, quality of family life

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Authors: D. Kuvshinov, A. Siswanto, W. Zimmerman

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A phorbol-12-myristate-13-acetate (TPA) is a synthetic analogue of phorbol ester (PE), a natural toxic compound of Euphorbiaceae plant. The oil extracted from plants of this family is useful source for primarily biofuel. However this oil can also be used as a food stock due to its significant nutrition content. The limitations for utilizing the oil as a food stock are mainly due to a toxicity of PE. Nowadays a majority of PE detoxification processes are expensive as include multi steps alcohol extraction sequence. Ozone is considered as a strong oxidative agent. It reaction with PE it attacks the carbon double bond of PE. This modification of PE molecular structure results into nontoxic ester with high lipid content. This report presents data on development of simple and cheap PE detoxification process with water application as a buffer and ozone as reactive component. The core of this new technique is a simultaneous application of new microscale plasma unit for ozone production and patented gas oscillation technology. In combination with a reactor design the technology permits ozone injection to the water-TPA mixture in form of microbubbles. The efficacy of a heterogeneous process depends on diffusion coefficient which can be controlled by contact time and interface area. The low velocity of rising microbubbles and high surface to volume ratio allow fast mass transfer to be achieved during the process. Direct injection of ozone is the most efficient process for a highly reactive and short lived chemical. Data on the plasma unit behavior are presented and influence of the gas oscillation technology to the microbubbles production mechanism has been discussed. Data on overall process efficacy for TPA degradation is shown.

Keywords: microbubble, ozonolysis, synthetic phorbol ester, chemical engineering

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Authors: Ballout Amor, Yasmina Bouchahm, Lacheheb Dhia Eddine Zakaria

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Several studies all around the world were conducted on the phenomenon of the urban heat island, and referring to the results obtained, one of the most important factors that influence this phenomenon is the mineralization of the cities which means the reducing of evaporative urban surfaces, replacing vegetation and wetlands with concrete and asphalt. The use of vegetation and water can change the urban environment and improve comfort, thus reduce the heat island. The trees act as a mask to the sun, wind, and sound, and also as a source of humidity which reduces air temperature and surrounding surfaces. Water also acts as a buffer to noise; it is also a source of moisture and regulates temperature not to mention the psychological effect on humans. Our main objective in this paper is to determine the impact of vegetation, ponds and fountains on the urban micro climate in general and on the thermal comfort of people along the Independence square in the Algerian city of Sétif, which is a semi-arid climate, in particularly. In order to reach this objective, a comparative study between different scenarios has been done; the use of the Envi-met program enabled us to model the urban environment of the Independence Square and to study the possibility of improving the conditions of comfort by adding an amount of vegetation and water ponds. After studying the results obtained (temperature, relative humidity, wind speed, PMV and PPD indicators), the efficiency of the additions we've made on the square was confirmed and this is what helped us to confirm our assumptions regarding the terms of comfort in the studied site, and in the end we are trying to develop recommendations and solutions which may contribute to improve the conditions for greater comfort in the Independence square.

Keywords: comfort in outer space, urban environment, scenarisation, vegetation, water ponds, public square, simulation

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Authors: Sahana Selladurai, Christine DeWolf

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Lung surfactant comprises a lipid-protein film that coats the alveolar surface and serves to prevent alveolar collapse upon repeated breathing cycles. Exposure of lung surfactant to high concentrations of airborne pollutants, for example tropospheric ozone in smog, can chemically modify the lipid and protein components. These chemical changes can impact the film functionality by decreasing the film’s collapse pressure (minimum surface tension attainable), altering it is mechanical and flow properties and modifying lipid reservoir formation essential for re-spreading of the film during the inhalation process. In this study, we use Langmuir monolayers spread at the air-water interface as model membranes where the compression and expansion of the film mimics the breathing cycle. The impact of ozone exposure on model lung surfactant films is measured using a Langmuir film balance, Brewster angle microscopy and a pendant drop tensiometer as a function of film and sub-phase composition. The oxidized films are analyzed using mass spectrometry where lipid and protein oxidation products are observed. Oxidation is shown to reduce surface activity, alter line tension (and film morphology) and in some cases visibly reduce the viscoelastic properties of the film when compared to controls. These reductions in functionality of the films are highly dependent on film and sub-phase composition, where for example, the effect of oxidation is more pronounced when using a physiologically relevant buffer as opposed to water as the sub-phase. These findings can lead to a better understanding on the impact of continuous exposure to high levels of ozone on the mechanical process of breathing, as well as understanding the roles of certain lung surfactant components in this process.

Keywords: lung surfactant, oxidation, ozone, viscoelasticity

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Authors: Jari Koskiaho, Teija Kirkkala, Jani Salminen, Sarianne Tikkanen, Sirkka Tattari

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Here, phosphorus (P) loading to the lake Pyhäjärvi (SW Finland) was reviewed, load reduction targets were determined, and different measures of waterwise circular economy to reach the targets were evaluated. In addition to the P loading from the lake’s catchment, there is a significant amount of internal P loading occurring in the lake. There are no point source emissions into the lake. Thus, the most important source of external nutrient loading is agriculture. According to the simulations made with LLR-model, the chemical state of the lake is at the border of the classes ‘Satisfactory’ and ‘Good’. The LLR simulations suggest that a reduction of some hundreds of kilograms in annual P loading would be needed to reach an unquestionably ‘Good’ state. Evaluation of the measures of the waterwise circular economy suggested that they possess great potential in reaching the target P load reduction. If they were applied extensively and in a versatile, targeted manner in the catchment, their combined effect would reach the target reduction. In terms of cost-effectiveness, the waterwise measures were ranked as follows: The best: Fishing, 2nd best: Recycling of vegetation of reed beds, wetlands and buffer zones, 3rd best: Recycling field drainage waters stored in wetlands and ponds for irrigation, 4th best: Controlled drainage and irrigation, and 5th best: Recycling of the sediments of wetlands and ponds for soil enrichment. We also identified various waterwise nutrient recycling measures to decrease the P content of arable land. The cost-effectiveness of such measures may be very good. Solutions are needed to Finnish water protection in general, and particularly for regions like lake Pyhäjärvi catchment with intensive domestic animal production, of which the ‘P-hotspots’ are a crucial issue.

Keywords: circular economy, lake protection, mitigation measures, phosphorus

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Authors: Hatem I. Mokhtar, Randa A. Abd-El-Salam, Ghada M. Hadad

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An improved nano-composite of self-doped polyaniline nanofibers and silica-coated magnetite nanoparticles were prepared and evaluated for suitability to magnetic dispersive micro solid-phase extraction. The work focused on optimization of the composite capacity to extract four fluoroquinolones (FQs) antibiotics, ciprofloxacin, enrofloxacin, danofloxacin, and difloxacin from water and improvement of composite stability towards acid and atmospheric degradation. Self-doped polyaniline nanofibers were prepared by oxidative co-polymerization of aniline with anthranilic acid. Magnetite nanopariticles were prepared by alkaline co-precipitation and coated with silica by silicate hydrolysis on magnetite nanoparticles surface at pH 6.5. The composite was formed by self-assembly by mixing self-doped polyaniline nanofibers with silica-coated magnetite nanoparticles dispersions in ethanol. The composite structure was confirmed by transmission electron microscopy (TEM). Self-doped polyaniline nanofibers and magnetite chemical structures were confirmed by FT-IR while silica coating of the magnetite was confirmed by Energy Dispersion X-ray Spectroscopy (EDS). Improved stability of the composite magnetic component was evidenced by resistance to degrade in 2N HCl solution. The adsorption capacity of self-doped polyaniline nanofibers based composite was higher than previously reported corresponding composite prepared from polyaniline nanofibers instead of self-doped polyaniline nanofibers. Adsorption-pH profile for the studied FQs on the prepared composite revealed that the best pH for adsorption was in range of 6.5 to 7. Best extraction recovery values were obtained at pH 7 using phosphate buffer. The best solvent for FQs desorption was found to be 0.1N HCl in methanol:water (8:2; v/v) mixture. 20 mL of Spiked water sample with studied FQs were preconcentrated using 4.8 mg of composite and resulting extracts were analysed by HPLC-UV method. The prepared composite represented a suitable adsorbent phase for magnetic dispersive micro-solid phase application.

Keywords: fluoroquinolones, magnetic dispersive micro extraction, nano-composite, self-doped polyaniline nanofibers

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Authors: Mohammed Ghazwani, Shyma Ali Alshahrani, Zahra Abdu Yousef, Taif Torki Asiri, Ghofran Abdur Rahman, Asma Ali Alshahrani, Umme Hani

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Background and introduction: Piroxicam is a nonsteroidal anti-inflammatory drug characterized by low solubility-high permeability used to reduce pain, swelling, and joint stiffness from arthritis. Hydrotropes are a class of compounds that normally increase the aqueous solubility of insoluble solutes. Aim: The objective of the present research study was to formulate and optimize Piroxicam hydrotropic starch gel using sodium salicylate, sodium benzoate as hydrotropic salts, and potato starch for topical application. Materials and methods: The prepared Piroxicam hydrotropic starch gel was characterized for various physicochemical parameters like drug content estimation, pH, tube extrudability, and spreadability; all the prepared formulations were subjected to in-vitro diffusion studies for six hours in 100 ml phosphate buffer (pH 7.4) and determined gel strength. Results: All formulations were found to be white opaque in appearance and have good homogeneity. The pH of formulations was found to be between 6.9-7.9. Drug content ranged from 96.8%-99.4.5%. Spreadability plays an important role in patient compliance and helps in the uniform application of gel to the skin as gels should spread easily; F4 showed a spreadability of 2.4cm highest among all other formulations. In in vitro diffusion studies, extrudability and gel strength were good with F4 in comparison with other formulations; hence F4 was selected as the optimized formulation. Conclusion: Isolated potato starch was successfully employed to prepare the gel. Hydrotropic salt sodium salicylate increased the solubility of Piroxicam and resulted in a stable gel, whereas the gel prepared using sodium benzoate changed its color after one week of preparation from white to light yellowish. Hydrotropic potato starch gel proposed a suitable vehicle for the topical delivery of Piroxicam.

Keywords: Piroxicam, potato starch, hydrotropic salts, hydrotropic starch gel

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Authors: Poonam Malik, Ravi Bhushan

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This work describes a very efficient approach for synthesis of activated ester of (S)-naproxen which was characterized by UV, IR, ¹HNMR, elemental analysis and polarimetric studies. It was used as a C-N bond forming chiral derivatizing reagent for further synthesis of diastereomeric amides of (RS)-salbutamol (a β₂ agonist that belongs to the group β-adrenolytic and is marketed as racamate) under microwave irradiation. The diastereomeric pair was separated by achiral phase HPLC, using mobile phase in gradient mode containing methanol and aqueous triethylaminephosphate (TEAP); separation conditions were optimized with respect to pH, flow rate, and buffer concentration and the method of separation was validated as per International Council for Harmonisation (ICH) guidelines. The reagent proved to be very effective for on-line sensitive detection of the diastereomers with very low limit of detection (LOD) values of 0.69 and 0.57 ng mL⁻¹ for diastereomeric derivatives of (S)- and (R)-salbutamol, respectively. The retention times were greatly reduced (2.7 min) with less consumption of organic solvents and large (α) as compared to literature reports. Besides, the diastereomeric derivatives were separated and isolated by preparative HPLC; these were characterized and were used as standard reference samples for recording ¹HNMR and IR spectra for determining absolute configuration and elution order; it ensured the success of diastereomeric synthesis and established the reliability of enantioseparation and eliminated the requirement of pure enantiomer of the analyte which is generally not available. The newly developed reagent can suitably be applied to several other amino group containing compounds either from organic syntheses or pharmaceutical industries because the presence of (S)-Npx as a strong chromophore would allow sensitive detection.This work is significant not only in the area of enantioseparation and determination of absolute configuration of diastereomeric derivatives but also in the area of developing new chiral derivatizing reagents (CDRs).

Keywords: chiral derivatizing reagent, naproxen, salbutamol, synthesis

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Authors: Victor S. Klimin, Alexey A. Rezvan, Maxim S. Solodovnik, Oleg A. Ageev

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In this paper, the problems of existing methods of profiling and surface modification of nanoscale arsenide-gallium structures are analyzed. The use of a combination of methods of local anodic oxidation and plasma chemical etching to solve this problem is considered. The main features that make this technology one of the promising areas of modification and profiling of near-surface layers of solids are demonstrated. In this paper, we studied the effect of formation stress and etching time on the geometrical parameters of the etched layer and the roughness of the etched surface. Experimental dependences of the thickness of the etched layer on the time and stress of formation were obtained. The surface analysis was carried out using atomic force microscopy methods, the corresponding profilograms were constructed from the obtained images, and the roughness of the etched surface was studied accordingly. It was shown that at high formation voltage, the depth of the etched surface increased, this is due to an increase in the number of active particles (oxygen ions and hydroxyl groups) formed as a result of the decomposition of water molecules in an electric field, during the formation of oxide nanostructures on the surface of gallium arsenide. Oxide layers were used as negative masks for subsequent plasma chemical etching by the STE ICPe68 unit. BCl₃ was chosen as the chlorine-containing gas, which differs from analogs in some parameters for the effect of etching of nanostructures based on gallium arsenide in the low-temperature plasma. The gas mixture of reaction chamber consisted of a buffer gas NAr = 100 cm³/min and a chlorine-containing gas NBCl₃ = 15 cm³/min at a pressure P = 2 Pa. The influence of these methods modes, which are formation voltage and etching time, on the roughness and geometric parameters, and corresponding dependences are demonstrated. Probe nanotechnology was used for surface analysis.

Keywords: nanostructures, GaAs, plasma chemical etching, modification structures

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Authors: Joseph A. Bentil, Anders Thygesen, Lene Langea, Moses Mensah, Anne Meyer

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The study investigated the saccharification potential of in-house enzymes produced from a white-rot basidiomycete strain, Polyporus ciliatus CBS 366.74. The in-house enzymes were produced by growing the fungus on mono and composite substrates of cocoa pod husk (CPH) and green seaweed (GS) (Ulva lactuca sp.) with and without the addition of 25mM ammonium nitrate at 4%w/v substrate concentration in submerged condition for 144 hours. The crude enzyme extracts preparations (CEE 1-5 and CEE 1-5+AN) obtained from the fungal cultivation process were sterile-filtered and used as enzyme sources for enzymatic hydrolysis of hydrothermally pretreated corn stover using a commercial cocktail enzyme, Cellic Ctec3, as benchmark. The hydrolysis was conducted at 50ᵒC with 50mM sodium acetate buffer, pH 5 based on enzyme dosages of 5 and 10 CMCase Units/g biomass at 1%w/v dry weight substrate concentration at time points of 6, 24, and 72 hours. The enzyme activity profile of the in-house enzymes varied among the growth substrates with the composite substrates (50-75% GS and AN inclusion), yielding better enzyme activities, especially endoglucanases (0.4-0.5U/mL), β-glucosidases (0.1-0.2 U/mL), and xylanases (3-10 U/mL). However, nitrogen supplementation had no significant effect on enzyme activities of crude extracts from 100% GS substituted substrates. From the enzymatic hydrolysis, it was observed that the in-house enzymes were capable of hydrolysing the pretreated corn stover at varying degrees; however, the saccharification yield was less than 10%. Consequently, theoretical glucose yield was ten times lower than Cellic Ctec3 at both dosage levels. There was no linear correlation between glucose yield and enzyme dosage for the in-house enzymes, unlike the benchmark enzyme. It is therefore recommended that the in-house enzymes are used to complement the dosage of commercial enzymes to reduce the cost of biomass saccharification.

Keywords: enzyme production, hydrolysis yield, feedstock, enzyme blend, Polyporus ciliatus

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Authors: Joseph F. Cabrera, Rachael Dela Cruz

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This study specifically examines patterns that define ethnoracially diverse neighborhoods. Ethnoracial diversity is important as it facilitates cross-racial interactions within neighborhoods which have been theorized to be associated with such outcomes as intergroup harmony, the reduction of racial and ethnic prejudice and discrimination, and increases in racial tolerance. Los Angeles (LA) is an ideal location to study ethnoracial spatial patterns as it is one of the most ethnoracially diverse cities in the world. A large influx of Latinos, as well as Asians, have contributed to LA’s urban landscape becoming increasingly diverse over several decades. Our dataset contains all census tracts in Los Angeles County in 2016 and incorporates Census and ACS demographic and spatial data. We quantify ethnoracial diversity using a derivative of Simpson’s Diversity Index and utilize this measure to test previous literature that suggests Latinos are one of the key drivers of changing ethnoracial spatial patterns in Los Angeles. Preliminary results suggest that there has been an overall increase in ethnoracial diversity in Los Angeles neighborhoods over the past sixteen years. Patterns associated with this trend include decreases in predominantly white and black neighborhoods, increases in predominantly Latino and Asian neighborhoods, and a general decrease in the white populations of the most diverse neighborhoods. A similar pattern is seen in neighborhoods with large Latino increases- a decrease in white population, but with an increase in Asian and black populations. We also found support for previous research that suggests increases in Latino and Asian populations act as a buffer, allowing for black population increases without a sizeable decrease in the white population. Future research is needed to understand the underlying causes involved in many of the patterns and trends highlighted in this study.

Keywords: race, race and interaction, racial harmony, social interaction

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Authors: Rui Wu

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In the volatile modern manufacturing environment, new orders randomly occur at any time, while the pre-emptive methods are infeasible. This leads to a real-time scheduling method that can produce a reasonably good schedule quickly. The dynamic Flexible Job Shop problem is an NP-hard scheduling problem that hybrid the dynamic Job Shop problem with the Parallel Machine problem. A Flexible Job Shop contains different work centres. Each work centre contains parallel machines that can process certain operations. Many algorithms, such as genetic algorithms or simulated annealing, have been proposed to solve the static Flexible Job Shop problems. However, the time efficiency of these methods is low, and these methods are not feasible in a dynamic scheduling problem. Therefore, a dynamic rule selection scheduling system based on the reinforcement learning method is proposed in this research, in which the dynamic Flexible Job Shop problem is divided into several parallel machine problems to decrease the complexity of the dynamic Flexible Job Shop problem. Firstly, the features of jobs, machines, work centres, and flexible job shops are selected to describe the status of the dynamic Flexible Job Shop problem at each decision point in each work centre. Secondly, a framework of reinforcement learning algorithm using a double-layer deep Q-learning network is applied to select proper composite dispatching rules based on the status of each work centre. Then, based on the selected composite dispatching rule, an available operation is selected from the waiting buffer and assigned to an available machine in each work centre. Finally, the proposed algorithm will be compared with well-known dispatching rules on objectives of mean tardiness, mean flow time, mean waiting time, or mean percentage of waiting time in the real-time Flexible Job Shop problem. The result of the simulations proved that the proposed framework has reasonable performance and time efficiency.

Keywords: dynamic scheduling problem, flexible job shop, dispatching rules, deep reinforcement learning

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Authors: Velasco-Rendón María Del Carmen, Cuéllar-Bermúdez Sara Paulina, Parra-Saldívar Roberto

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Phycocyanin is a blue pigment protein with fluorescent activity produced by cyanobacteria. It has been recently studied to determine its anticancer, antioxidant and antiinflamatory potential. Since 2014 it was approved as a Generally Recognized As Safe (GRAS) proteic pigment for the food industry. Therefore, phycocyanin shows potential for the food, nutraceutical, pharmaceutical and diagnostics industry. Conventional phycocyanin extraction includes buffer solutions and ammonium sulphate followed by chromatography or ATPS for protein separation. Therefore, further purification steps are time-requiring, energy intensive and not suitable for scale-up processing. This work presents an alternative to conventional methods that also allows large scale application with commercially available equipment. The extraction was performed by exposing the dry biomass to mechanical cavitation and salting out with NaCl to use an edible reagent. Also, isoelectric point precipitation was used by addition of HCl and neutralization with NaOH. The results were measured and compared in phycocyanin concentration, purity and extraction yield. Results showed that the best extraction condition was the extraction by salting out with 0.20 M NaCl after 30 minutes cavitation, with a concentration in the supernatant of 2.22 mg/ml, a purity of 3.28 and recovery from crude extract of 81.27%. Mechanical cavitation presumably increased the solvent-biomass contact, making the crude extract visibly dark blue after centrifugation. Compared to other systems, our process has less purification steps, similar concentrations in the phycocyanin-rich fraction and higher purity. The contaminants present in our process edible NaCl or low pHs that can be neutralized. It also can be adapted to a semi-continuous process with commercially available equipment. This characteristics make this process an appealing alternative for phycocyanin extraction as a pigment for the food industry.

Keywords: extraction, phycocyanin, precipitation, scale-up

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Authors: Eftychia Pappa, Heleni Vastardis, Christos Rahiotis, Andriani Vazaiou

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The aim of this study was to evaluate the prevalence of dry-mouth symptoms (xerostomia) and compare it with alterations in salivary characteristics of children and adolescents with type 1 diabetes (DM1), as measured with the use of chair-side saliva tests. This study also investigated the possible association between salivary dysfunction and incidence of caries, in relation to the level of metabolic control. A cross-sectional study was performed on young patients (6-18 years old) allocated among 3 groups: 40 patients poorly-controlled (DM1-A, HbA1c>8%), 40 well-controlled (DM1-B, HbA1c≤8%) and 40 age- and sex-matched healthy controls. The study was approved by the Research Ethics Committee of University of Athens and the parents signed written informed consent. All subjects were examined for dental caries, oral hygiene and salivary factors. Assessments of salivary function included self-reported xerostomia, quantification of resting and stimulated whole saliva flow rates, pH values, buffering capacity and saliva’s viscosity. Salivary characteristics were evaluated with the use of GC Saliva Check Buffer (3Μ ESPE). Data were analysed by chi-square and Kruskal-Wallis tests. Subjects with diabetes reported xerostomia more frequently than healthy controls (p<0.05). Unstimulated salivary flow rate and pH values remained significantly lower in DM1-A compared to DM1-B and controls. Low values of resting salivary flow rate were associated with a higher prevalence of dental caries in children and adolescents with poorly-controlled DM1 (p<0.05). The results suggested that diabetes-induced alterations in salivary characteristics are indicative of higher caries susceptibility of diabetics and chair-side saliva tests are a useful tool for the evaluation of caries risk assessment.

Keywords: caries risk assessment, saliva diagnostic tests, type 1 diabetes, xerostomia

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Authors: G. Nampa, M. Ndlovu

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Mutualistic relationships amongst organisms drive diversity in terrestrial ecosystems. Yet, few mutual associations have been documented in the semi-arid savannas of Africa. The levels and benefits of association between Carissa bispinosa, a medium-sized evergreen thorny shrub, and Trinervitermes trinervoides, an ecosystem engineer termite species, were studied at a semi-arid savanna setting in Nylsvley nature reserve, South Africa. It was hypothesized that there would be a close plant-insect association since termite mounds provide nutrients for plant growth and, in return, the thorny shrubs protect mounds from predation and also provide a temperature buffer. Comparative plant and mounds measurements were taken from associated and isolated occurrences seasonally. Soil particle size, macro- and micronutrients were also evaluated from mounds and the adjacent topsoil matrix General Additive Mixed Models were used to assess internal mound temperatures in relation to prevailing ambient and plant shade temperatures. Findings revealed that plants growing on mounds were significantly taller with a wider canopy and remained greener in the dry season with more fruits. On the other hand, termite mounds under plants were less prone to be damaged by aardvarks and pangolins and had a significantly wider diameter than exposed mounds. All soil macronutrients except for calcium and phosphorous were enriched in mounds relative to the matrix. Only Manganese was enriched in mounds while the other micronutrients (Cu, Fe, Zn and B) were not. Termite mounds under plants maintained a better constant and higher mean internal temperature during winter compared to exposed mounds. To our best knowledge, the study has revealed a previously undocumented survival mechanism that termites use to escape extreme temperatures and predation in semi-arid savannas.

Keywords: mound, mutualism, soil nutrients, termites, thermoregulation

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Authors: Shringika Soni, Utkarsh Jain, Nidhi Chauhan

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Aptamers are recently discovered ~80-100 bp long artificial oligonucleotides that not only demonstrated their applications in therapeutics; it is tremendously used in diagnostic and sensing application to detect different biomarkers and drugs. Synthesizing aptamers for proteins or genomic template is comparatively feasible in laboratory, but drugs or other chemical target based aptamers require major specification and proper optimization and validation. One has to optimize all selection, amplification, and characterization steps of the end product, which is extremely time-consuming. Therefore, we performed asymmetric PCR (polymerase chain reaction) for random oligonucleotides pool synthesis, and further use them in Systematic evolution of ligands by exponential enrichment (SELEX) for anti-psychotic drugs based aptamers synthesis. Anti-psychotic drugs are major tranquilizers to control psychosis for proper cognitive functions. Though their low medical use, their misuse may lead to severe medical condition as addiction and can promote crime in social and economical impact. In this work, we have approached the in-vitro SELEX method for ssDNA synthesis for anti-psychotic drugs (in this case ‘target’) based aptamer synthesis. The study was performed in three stages, where first stage included synthesis of random oligonucleotides pool via asymmetric PCR where end product was analyzed with electrophoresis and purified for further stages. The purified oligonucleotide pool was incubated in SELEX buffer, and further partition was performed in the next stage to obtain target specific aptamers. The isolated oligonucleotides are characterized and quantified after each round of partition, and significant results were obtained. After the repetitive partition and amplification steps of target-specific oligonucleotides, final stage included sequencing of end product. We can confirm the specific sequence for anti-psychoactive drugs, which will be further used in diagnostic application in clinical and forensic set-up.

Keywords: anti-psychotic drugs, aptamer, biosensor, ssDNA, SELEX

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Authors: Luojie Tang

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Riverside space is an important public space and ecological corridor in urban areas, but mountainous urban rivers are often overlooked due to their deep valleys and poor accessibility. This article takes the Qing Shui Xi River in Chongqing as an example, and through long-term field inspections, measurements, interviews, and online surveys, summarizes the problems of poor accessibility, limited space for renovation, lack of waterfront facilities, excessive artificial intervention, low average runoff, severe river water pollution, and difficulty in integrated watershed management in riverside space. Based on the current situation and drawing on relevant experiences, this article summarizes the design methods for riverside space in deep valley rivers in mountainous urban areas. Regarding spatial design techniques, the article emphasizes the importance of integrating waterfront spaces into the urban public space system and vertical linkages. Furthermore, the article suggests different design methods and improvement strategies for the already developed areas and new development areas. Specifically, the article proposes a planning and design strategy of "protection" and "empowerment" for new development areas and an updating and transformation strategy of "improvement" and "revitalization" for already developed areas. In terms of ecological restoration methods, the article suggests three focus points: increasing the runoff of urban rivers, raising the landscape water level during dry seasons, and restoring vegetation and wetlands in the riverbank buffer zone while protecting the overall pattern of the watershed. Additionally, the article presents specific design details of the Qingshuixi River to illustrate the proposed design and restoration techniques.

Keywords: deep-cut river, design method, mountainous city, Qingshuixi river in Chongqing, waterfront space design

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Authors: Sidra Pervez, Afsheen Aman, Shah Ali Ul Qader

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The importance of attaining an optimum performance of an enzyme is often a question of devising an effective method for its immobilization. Cross-linked enzyme aggregate (CLEAs) is a new approach for immobilization of enzymes using carrier free strategy. This method is exquisitely simple (involving precipitation of the enzyme from aqueous buffer followed by cross-linking of the resulting physical aggregates of enzyme molecules) and amenable to rapid optimization. Among many industrial enzymes, amyloglucosidase is an important amylolytic enzyme that hydrolyzes alpha (1→4) and alpha (1→6) glycosidic bonds in starch molecule and produce glucose as a sole end product. Glucose liberated by amyloglucosidase can be used for the production of ethanol and glucose syrups. Besides this amyloglucosidase can be widely used in various food and pharmaceuticals industries. For production of amyloglucosidase on commercial scale, filamentous fungi of genera Aspergillus are mostly used because they secrete large amount of enzymes extracellularly. The current investigation was based on isolation and identification of filamentous fungi from genus Aspergillus for the production of amyloglucosidase in submerged fermentation and optimization of cultivation parameters for starch saccharification. Natural isolates were identified as Aspergillus niger KIBGE-IB36, Aspergillus fumigatus KIBGE-IB33, Aspergillus flavus KIBGE-IB34 and Aspergillus terreus KIBGE-IB35 on taxonomical basis and 18S rDNA analysis and their sequence were submitted to GenBank. Among them, Aspergillus fumigatus KIBGE-IB33 was selected on the basis of maximum enzyme production. After optimization of fermentation conditions enzyme was immobilized on CLEA. Different parameters were optimized for maximum immobilization of amyloglucosidase. Data of enzyme stability (thermal and Storage) and reusability suggested the applicability of immobilized amyloglucosidase for continuous saccharification of starch in industrial processes.

Keywords: aspergillus, immobilization, industrial processes, starch saccharification

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Authors: Farhad Reyazat, Richard Werner

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This paper contributes to the existent literature by developing a framework that explains how to monitor potential threats to banking sector stability. The study explores structural vulnerabilities at the country level, but also look at bilateral exposures within a network context. The study contributes in analysing of the European banking systemic risk at aggregated level, which integrates the characteristics of bank size, and interconnectedness relative to the size of the economy which ultimate risk belong to, taking to account the concentration ratio of the banking industry within the whole economy. The nature of the systemic risk depends on the interplay of the network topology with the nature of financial transactions over the network, assets and buffer stemming from bank size, correlations, and the nature of the shocks to the financial system. The study’s results illustrate the contribution of banks’ size, size of economy and concentration of counterparty exposures to a given country’s banks in explaining its systemic importance, how much the banking network depends on a few traditional hubs activities and the changes of this dependencies over the last 9 years. The role of few of traditional hubs such as Swiss banks and British Banks and also Irish banks- where the financial sector is fairly new and grew strongly between 1990s till 2008- take the fourth position on 2014 reducing the relative size since 2006 where they had the first position. In-degree concentration index analysis in the study shows concentration index of banking network was not changed since financial crisis 2007-8. In-degree concentration index on first quarter of 2014 indicates that US, UK and Germany together, getting over 70% of the network exposures. The result of comparing the in-degree concentration index with 2007-4Q, shows the same group having over 70% of the network exposure, however the UK getting more important role in the hub and the market share of US and Germany are slightly diminished.

Keywords: systemic risk, counterparty risk, financial stability, interconnectedness, banking concentration, european banks risk, network effect on systemic risk, concentration risk

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Authors: Dawit Assefa, Kassaye Tekie, Gebrie Alebachew, Degefu Beyene, Bikila Alemu, Naji Mohammed, Asnakech Agegnehu, Seble Tsehay, Geremew Tasew

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Background: Implementation of a Laboratory Quality Management System (LQMS) is critical to ensure accurate, reliable, and efficient laboratory testing of antimicrobial resistance (AMR). However, limited LQMS implementation and progress toward accreditation in the AMR surveillance laboratory testing setting exist in Ethiopia. By addressing non-conformances (NCs) and working towards accreditation, microbiology laboratories can improve the quality of their services, increase staff competence, and contribute to mitigate the spread of AMR. Methods: Using standard ISO 15189 horizontal and vertical assessment checklists, certified assessors identified NCs at Hawassa and Jimma Hospital microbiology laboratories. The Ethiopian Public Health Institute AMR mentors and IDDS staff prioritized closing the NCs through the implementation of an intensified mentorship program that included ISO 15189 orientation training, resource allocation, and action plan development. Results: For the two facilities to clear their NCs, an intensified mentorship approach was adopted by providing ISO 15189 orientation training, provision of buffer reagents, controls, standards, and axillary equipment, and facilitating equipment maintenance and calibration. Method verification and competency assessment were also conducted along with the implementation of standard operating procedures and recommended corrective actions. This approach enhanced the laboratory's readiness for accreditation. After addressing their NCs, the two laboratories applied to Ethiopian Accreditation Services for ISO 15189 accreditation. Conclusions: Clearing NCs through the implementation of intensified mentorship was crucial in preparing the two laboratories for accreditation and improving quality laboratory test results. This approach can guide other microbiology laboratories’ accreditation attainment efforts.

Keywords: non-conformance clearance, intensified mentorship, accreditation, ISO 15189

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Authors: Bea Carmel H. Casiding, Charmaine A. Guy, Funny Jovis P. Malasan, Katrina Chelsea B. Manlutac, Danielle Ann N. Novilla, Marianne R. Oliveros, Magnolia C. Sibulo

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Moon jellyfish, Aurelia aurita, has become a popular research organism for diverse studies. Recent studies have verified the prevention of blood clotting properties of the moon jellyfish tentacle extract through in vitro methods. The purpose of this study was to validate the blood clotting ability of A. aurita tentacle extract using in vivo method of experimentation. The tentacles of A. aurita jellyfish were excised and filtered then centrifuged at 3000xg for 10 minutes. The crude nematocyst extract was suspended in 1:6 ratios with phosphate buffer solution and sonicated for three periods of 20 seconds each at 50 Hz. Protein concentration of the extract was determined using Bradford Assay. Bovine serum albumin was the standard solution used with the following concentrations: 35.0, 70.0, 105.0, 140.0, 175.0, 210.0, 245.0, and 280.0 µg/mL. The absorbance was read at 595 nm. Toxicity testing from OECD guidelines was adapted. The extract suspended in phosphate-buffered saline solution was arbitrarily set into three doses (0.1mg/kg, 0.3mg/kg, 0.5mg/kg) and were administered daily for five days to the experimental groups of five male Sprague-Dawley rats (one dose per group). Before and after the administration period, bleeding time and clotting time tests were performed. The One-way Analysis of Variance (ANOVA) was used to analyze the difference of before and after bleeding time and clotting time from the three treatment groups, time, positive and negative control groups. The average protein concentration of the sonicated crude tentacle extract was 206.5 µg/mL. The highest dose administered (0.5mg/kg) produced significant increase in the time for both bleeding and clotting tests. However, the preceding lower dose (0.3mg/kg) only was significantly effective for clotting time test. The protein contained in the tentacle extract with a concentration of 206.5 mcg/mL and dose of 0.3 mg/kg and 0.5 mg/kg of A. aurita elicited anticoagulating activity.

Keywords: anticoagulant, bleeding time test, clotting time test, moon jellyfish

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Authors: Haiming Wen, Isabella J. Van Rooyen

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Tristructural isotropic (TRISO)-coated fuel particles are designed as nuclear fuel for high-temperature gas reactors. TRISO coating consists of layers of carbon buffer, inner pyrolytic carbon (IPyC), SiC, and outer pyrolytic carbon. The TRISO coating, especially the SiC layer, acts as a containment system for fission products produced in the kernel. However, release of certain metallic fission products across intact TRISO coatings has been observed for decades. Despite numerous studies, mechanisms by which fission products migrate across the coating layers remain poorly understood. In this study, scanning transmission electron microscopy (STEM), energy dispersive X-ray spectroscopy (EDS), high-resolution transmission electron microscopy (HRTEM) and electron energy loss spectroscopy (EELS) were used to examine the distribution, composition and structure of fission products in a TRISO coated particle neutron irradiated to 3.6 x 1021 n/cm² fast fluence at 1040 ⁰C. Precession electron diffraction was used to investigate characters of grain boundaries where specific fission product precipitates are located. The retention fraction of 110mAg in the investigated TRISO particle was estimated to be 0.19. A high density of nanoscale fission product precipitates was observed in the SiC layer close to the SiC-IPyC interface, most of which are rich in Pd, while Ag was not identified. Some Pd-rich precipitates contain U. Precipitates tend to have complex structure and composition. Although a precipitate appears to have uniform contrast in STEM, EDS indicated that there may be composition variations throughout the precipitate, and HRTEM suggested that the precipitate may have several parts different in crystal structure or orientation. Attempts were made to measure charge states of precipitates using EELS and study their possible effect on precipitate transport.

Keywords: TRISO particle, fission product, nuclear fuel, electron microscopy, neutron irradiation

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Authors: E. N. Vetrova, L.V. Lapochkina, N. V. Nikulina

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In the national economy, entrepreneurship plays the role of a buffer between economy and policy for it contributes to improving budget effectiveness and decreasing dependence of economy on the state. Entrepreneurship in industry makes it possible to increase the added value that is formed in production chains and to decrease dependence on import. Under the current circumstances, when sanctions are being imposed, this is especially relevant for Russia and for the realization of projects in the Russian Arctic. However, development of entrepreneurship in industry requires an enlightened state policy. The purpose of the research is elaboration of recommendations for improving economic effectiveness of the realization of the Arctic projects on the basis of conceptual proposals for the development of entrepreneurship in industry. The paper presents the studies of the extractive industry role in the Russian economy and proves its raw material character. The analysis of production chains in industry on the basis of the conception of the added value global chains demonstrated a low added value formed by Russian companies. The study of changes in the structure of economy based on systemic, statistical and comparative analyses revealed no positive changes in the structure of economy over the period under consideration. This is a manifestation of ineffectiveness of the Russian industrial policy in general and within the Arctic region in particular. The authors identified the problems information and implementation of the state industrial policy in the Arctic region and in the development of national entrepreneurship, analyzed the shortcomings of the current state policy in the sphere of the Russian industry. On the basis of the conducted studies, the authors formulated conceptual approaches to change the state policy in the Arctic. The basic idea of the authors is to substantiate the focus of the state regulation on the development of entrepreneurship in industry in the process of the Russian Arctic exploration. At the same time another problem is solved–that of the development of the manufacturing industry in the southern regions of the northwestern part of Russia. The criterion of effectiveness in this case is the economic effectiveness.

Keywords: entrepreneurship in industry, global chains of the added value, government regulation, industrial policies, production chains in the arctic region, economic effectiveness

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Authors: Grace E. Santella, Shawn G. Gallaher, Joseph P. Smith

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In order to determine the importance of small-system riverine heat flux on regional landfast sea ice breakup, our study explores the annual spring freshet of the Sagavanirktok River from 2014-2019. Seasonal heat cycling ultimately serves as the driving mechanism behind the freshet; however, as an emerging area of study, the extent to which inland thermodynamics influence coastal tundra geomorphology and connected landfast sea ice has not been extensively investigated in relation to small-scale Arctic river systems. The Sagavanirktok River is a small-to-midsized river system that flows south-to-north on the Alaskan North Slope from the Brooks mountain range to the Beaufort Sea at Prudhoe Bay. Seasonal warming in the spring rapidly melts snow and ice in a northwards progression from the Brooks Range and transitional tundra highlands towards the coast and when coupled with seasonal precipitation, results in a pulsed freshet that propagates through the Sagavanirktok River. The concentrated presence of newly exposed vegetation in the transitional tundra region due to spring melting results in higher absorption of solar radiation due to a lower albedo relative to snow-covered tundra and/or landfast sea ice. This results in spring flood runoff that advances over impermeable early-season permafrost soils with elevated temperatures relative to landfast sea ice and sub-ice flow. We examine the extent to which interannual temporal variability influences the onset and magnitude of river discharge by analyzing field measurements from the United States Geological Survey (USGS) river and meteorological observation sites. Rapid influx of heat to the Arctic Ocean via riverine systems results in a noticeable decay of landfast sea ice independent of ice breakup seaward of the shear zone. Utilizing MODIS imagery from NASA’s Terra satellite, interannual variability of river discharge is visualized, allowing for optical validation that the discharge flow is interacting with landfast sea ice. Thermal erosion experienced by sediment fast ice at the arrival of warm overflow preconditions the ice regime for rapid thawing. We investigate the extent to which interannual heat flux from the Sagavanirktok River’s freshet significantly influences the onset of local landfast sea ice breakup. The early-season warming of atmospheric temperatures is evidenced by the presence of storms which introduce liquid, rather than frozen, precipitation into the system. The resultant decreased albedo of the transitional tundra supports the positive relationship between early-season precipitation events, inland thermodynamic cycling, and degradation of landfast sea ice. Early removal of landfast sea ice increases coastal erosion in these regions and has implications for coastline geomorphology which stress industrial, ecological, and humanitarian infrastructure.

Keywords: Albedo, freshet, landfast sea ice, riverine heat flux, seasonal heat cycling

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Authors: Endalew Jibat, Feyera Senbeta, Tesfaye Zeleke, Fitsum Hagos

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Water governance considers multi-sector participation beyond the state; and for sustainable use of water resources, appropriate laws, policies, regulations, and institutions needs to be developed and put in place. Water policy, a critical and integral instrument of water governance, guided water use schemes and ensures equitable water distribution among users. The Ethiopian Central Rift Valley (CRV) is wealthy of water resources, but these water resources are currently under severe strain owing to an imbalance in human-water interactions. The main aim of the study was to examine the state of water resources governance in the CRV of Ethiopia, and the impact of the Ethiopian Water Resources Management Policy on water governance. Key informant interviews (KII), focused group discussions, and document reviews were used to gather data for the study. The NVivo 11 program was used to organize, code, and analyze the data. The results revealed that water resources governance practices such as water allocation and apportionment, comprehensive and integrated water management plans, water resources protection, and conservation activities were rarely implemented. Water resources management policy mechanisms were not fully put in place. Lack of coherence in water policy implementation, absence of clear roles and responsibilities of stakeholders, absence of transparency and accountability in irrigation water service delivery, and lack of meaningful participation of key actors in water governance decision-making were the primary shortcomings observed. Factors such as over-abstraction, deterioration of buffer zone, and chemical erosion from surrounding farming have contributed to the reduction in water volume and quality in the CRV. These challenges have influenced aquatic ecosystem services and threaten the livelihoods of the surrounding communities. Hence, reforms relating to policy coherence and enforcement, stakeholder involvement, water distribution strategies, and the application of water governance principles must be given more emphasis.

Keywords: water resources, irrigation, governance, water allocation, governance principles, stakeholders engagement, central rift valley

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Authors: Eylem G. Cengiz, Ersin Çilek, Gözde Başkaya, Havva Nur Taş

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It is possible to define the abuse and neglect of the child as a social problem. Such adverse experiences of the child are witnessed by wider social circles as well as his or her immediate environment. The most effective institution among these social circles is the school. The awareness of teachers, administrators, and even auxiliary personnel on this issue can act as a protective and preventive buffer because teachers have the opportunity to be with children every day and constantly observe them; therefore, they can notice the physical and mental changes in students. Furthermore, school administrators have an undeniable role in recognizing and responding to the risk of neglect and abuse. This study aims to evaluate the awareness of school administrators on the risk factors, clues, and ways of intervention towards abuse and neglect through the scenarios given to them. Data were collected from 37 primary, secondary, and high school administrators working in various provinces of Turkey through descriptive and scenario-based questions to determine their general knowledge of the concepts of neglect and abuse and their general tendencies towards practice. Descriptive questions were evaluated with content analysis, and scenario-based questions were evaluated with numerical qualitative data analysis. Concepts and themes were tried to be reached by content analysis from the descriptive data collected. When the results are evaluated in general, it is striking that the concept of child abuse means only sexual abuse for some school administrators. There is an important uncertainty for school administrators in the content of the idea of neglect. When the views on the causes of neglect and abuse are examined, the family factor was seen as the primary source of both neglect and abuse. In addition, among the prevention strategies applied by school administrators, intervention for the family -interviewing and informing- was recommended by only 9 (29%) out of 31 administrators. When the responses to the physical, emotional, and sexual abuse scenarios are examined, it is revealed that the administrators generally realize the abuse but fail to develop an appropriate/whole intervention method. The research results show that school administrators' awareness should be increased. Although administrators have sensitivity towards children, they should be empowered to recognize all types of neglect and abuse and develop appropriate intervention tools.

Keywords: assessment child abuse and neglect, child abuse, child neglect, school administrators

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Authors: Anil Koklu, Amin Mansoorifar, Ali Beskok

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Dielectric spectroscopy (DS) is a noninvasive, label free technique for a long term real-time measurements of the impedance spectra of biological cells. DS enables characterization of cellular dielectric properties such as membrane capacitance and cytoplasmic conductivity. We have developed a lab-on-a-chip device that uses an electro-activated microwells array for loading, DS measurements, and unloading of biological cells. We utilized from dielectrophoresis (DEP) to capture target cells inside the wells and release them after DS measurement. DEP is a label-free technique that exploits differences among dielectric properties of the particles. In detail, DEP is the motion of polarizable particles suspended in an ionic solution and subjected to a spatially non-uniform external electric field. To the best of our knowledge, this is the first microfluidic chip that combines DEP and DS to analyze biological cells using electro-activated wells. Device performance is tested using two different cell lines of prostate cancer cells (RV122, PC-3). Impedance measurements were conducted at 0.2 V in the 10 kHz to 40 MHz range with 6 s time resolution. An equivalent circuit model was developed to extract the cell membrane capacitance and cell cytoplasmic conductivity from the impedance spectra. We report the time course of the variations in dielectric properties of PC-3 and RV122 cells suspended in low conductivity medium (LCB), which enhances dielectrophoretic and impedance responses, and their response to sudden pH change from a pH of 7.3 to a pH of 5.8. It is shown that microfluidic chip allowed online measurements of dielectric properties of prostate cancer cells and the assessment of the cellular level variations under external stimuli such as different buffer conductivity and pH. Based on these data, we intend to deploy the current device for single cell measurements by fabricating separately addressable N × N electrode platforms. Such a device will allow time-dependent dielectric response measurements for individual cells with the ability of selectively releasing them using negative-DEP and pressure driven flow.

Keywords: microfluidic, microfabrication, lab on a chip, AC electrokinetics, dielectric spectroscopy

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Authors: Yenny Paola Morales Cortés, Fabián Rico Rodríguez, Juan Carlos Serrato Bermúdez, Carlos Arturo Martínez Riascos

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Numerous benefits of galactooligosaccharides (GOS) as prebiotics have motivated the study of enzymatic processes for their production. These processes have special complexities due to several factors that make difficult high productivity, such as enzyme type, reaction medium pH, substrate concentrations and presence of inhibitors, among others. In the present work the production of galactooligosaccharides (with different degrees of polymerization: two, three and four) from lactose was studied. The study considers the formulation of a mathematical model that predicts the production of GOS from lactose using the enzyme β-galactosidase. The effect of pH in the reaction was studied. For that, phosphate buffer was used and with this was evaluated three pH values (6.0.6.5 and 7.0). Thus it was observed that at pH 6.0 the enzymatic activity insignificant. On the other hand, at pH 7.0 the enzymatic activity was approximately 27 times greater than at 6.5. The last result differs from previously reported results. Therefore, pH 7.0 was chosen as working pH. Additionally, the enzyme concentration was analyzed, which allowed observing that the effect of the concentration depends on the pH and the concentration was set for the following studies in 0.272 mM. Afterwards, experiments were performed varying the lactose concentration to evaluate its effects on the process and to generate the data for the adjustment of the mathematical model parameters. The mathematical model considers the reactions of lactose hydrolysis and transgalactosylation for the production of disaccharides and trisaccharides, with their inverse reactions. The production of tetrasaccharides was negligible and, because of that, it was not included in the model. The reaction was monitored by HPLC and for the quantitative analysis of the experimental data the Matlab programming language was used, including solvers for differential equations systems integration (ode15s) and nonlinear problems optimization (fminunc). The results confirm that the transgalactosylation and hydrolysis reactions are reversible, additionally inhibition by glucose and galactose is observed on the production of GOS. In relation to the production process of galactooligosaccharides, the results show that it is necessary to have high initial concentrations of lactose considering that favors the transgalactosylation reaction, while low concentrations favor hydrolysis reactions.

Keywords: β-galactosidase, galactooligosaccharides, inhibition, lactose, Matlab, modeling

Procedia PDF Downloads 324