Search results for: active surveillance

Authors: Ali Ahmed Ali Al-Juboori, H. Zhu, D. Wexler, H. Li, C. Lu, J. McLeod, S. Pannila, J. Barnes

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A number of studies have focused on the formation mechanics of white etching layer and its origin in the railway operation. Until recently, the following hypotheses consider the precise mechanics of WELs formation: (i) WELs are the result of thermal process caused by wheel slip; (ii) WELs are mechanically induced by severe plastic deformation; (iii) WELs are caused by a combination of thermo-mechanical process. The mechanisms discussed above lead to occurrence of white etching layers on the area of wheel and rail contact. This is because the contact patch which is the active point of the wheel on the rail is exposed to highest shear stresses which result in localised severe plastic deformation; and highest rate of heat caused by wheel slipe during excessive traction or braking effort. However, if the WELs are not on the running band area, it would suggest that there is another cause of WELs formation. In railway system, particularly electrified railway, arcing phenomenon has been occurring more often and regularly on the rails. In electrified railway, the current is delivered to the train traction motor via contact wires and then returned to the station via the contact between the wheel and the rail. If the contact between the wheel and the rail is temporarily losing, due to dynamic vibration, entrapped dirt or water, lubricant effect or oxidation occurrences, high current can jump through the gap and results in arcing. The other resources of arcing also include the wheel passage the insulated joint and lightning on a train during bad weather. During the arcing, an extensive heat is generated and speared over a large area of top surface of rail. Thus, arcing is considered another heat source in the rail head (rather than wheel slipe) that results in microstructural changes and white etching layer formation. A head hardened (HH) rail steel, cut from a curved rail truck was used for the investigation. Samples were sectioned from a depth of 10 mm below the rail surface, where the material is considered to be still within the hardened layer but away from any microstructural changes on the top surface layer caused by train passage. These samples were subjected to electrical discharges by using Gas Tungsten Arc Welding (GTAW) machine. The arc current was controlled and moved along the samples surface in the direction of travel, as indicated by an arrow. Five different conditions were applied on the surface of the samples. Samples containing pre-existed WELs, taken from ex-service rail surface, were also considered in this study for comparison. Both simulated and ex-serviced WELs were characterised by advanced methods including SEM, TEM, TKD, EDS, XRD. Samples for TEM and TKFD were prepared by Focused Ion Beam (FIB) milling. The results showed that both simulated WELs by electrical arcing and ex-service WEL comprise similar microstructure. Brown etching layer was found with WELs and likely induced by a concurrent tempering process. This study provided a clear understanding of new formation mechanics of WELs which contributes to track maintenance procedure.

Keywords: white etching layer, arcing, brown etching layer, material characterisation

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Authors: O. Gsib, U. Peirera, C. Egles, S. A. Bencherif

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In skin regenerative medicine, one of the critical issues is to produce a three-dimensional scaffold with optimized porosity for dermal fibroblast infiltration and neovascularization, which exhibits high mechanical properties and displays sufficient wound healing characteristics. In this study, we report on the synthesis and characterization of macroporous sequential interpenetrating polymer networks (IPNs) combining skin wound healing properties of fibrin with the excellent physical properties of polyethylene glycol (PEG). Fibrin fibers serve as a provisional biologically active network to promote cell adhesion and proliferation while PEG provides the mechanical stability to maintain the entire 3D construct. After having modified both PEG and Serum Albumin (used for promoting enzymatic degradability) by adding methacrylate residues (PEGDM and SAM, respectively), Fibrin/PEGDM-SAM sequential IPNs were synthesized as follows: Macroporous sponges were first produced from PEGDM-SAM hydrogels by a freeze-drying technique and then rehydrated by adding the fibrin precursors. Environmental Scanning Electron Microscopy (ESEM) and Confocal Laser Scanning Microscopy (CLSM) were used to characterize their microstructure. Human dermal fibroblasts were cultivated during one week in the constructs and different cell culture parameters (viability, morphology, proliferation) were evaluated. Subcutaneous implantations of the scaffolds were conducted on five-week old male nude mice to investigate their biocompatibility in vivo. We successfully synthesized interconnected and macroporous Fibrin/PEGDM-SAM sequential IPNs. The viability of primary dermal fibroblasts was well maintained (above 90%) after 2 days of culture. Cells were able to adhere, spread and proliferate in the scaffolds suggesting the suitable porosity and intrinsic biologic properties of the constructs. The fibrin network adopted a spider web shape that covered partially the pores allowing easier cell infiltration into the macroporous structure. To further characterize the in vitro cell behavior, cell proliferation (EdU incorporation, MTS assay) is being studied. Preliminary histological analysis of animal studies indicated the persistence of hydrogels even after one-month post implantation and confirmed the absence of inflammation response, good biocompatibility and biointegration of our scaffolds within the surrounding tissues. These results suggest that our Fibrin/PEGDM-SAM IPNs could be considered as potential candidates for dermis regenerative medicine. Histological analysis will be completed to further assess scaffold remodeling including de novo extracellular matrix protein synthesis and early stage angiogenesis analysis. Compression measurements will be conducted to investigate the mechanical properties.

Keywords: fibrin, hydrogels for dermal reconstruction, polyethylene glycol, semi-interpenetrating polymer network

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Authors: Benjamin Glasner, Carlos Henriquez, Fernando Alfaro, Nicole Trefault, Santiago Andrade, Rodrigo De La Iglesia

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A central question in ecology refers to the relative importance that local-scale variables have over community composition, when compared with regional-scale variables. In coastal environments, strong seasonal abiotic influence dominates these systems, weakening the impact of other parameters like micronutrients. After the industrial revolution, micronutrients like trace metals have increased in ocean as pollutants, with strong effects upon biotic entities and biological processes in coastal regions. Coastal picoplankton communities had been characterized as a cyanobacterial dominated fraction, but in recent years the eukaryotic component of this size fraction has gained relevance due to their high influence in carbon cycle, although, diversity patterns and responses to disturbances are poorly understood. South Pacific upwelling coastal environments represent an excellent model to study seasonal changes due to a strong influence in the availability of macro- and micronutrients between seasons. In addition, some well constrained coastal bays of this region have been subjected to strong disturbances due to trace metal inputs. In this study, we aim to compare the influence of seasonality and trace metals concentrations, on the community structure of planktonic picoeukaryotes. To describe seasonal patterns in the study area, satellite data in a 6 years time series and in-situ measurements with a traditional oceanographic approach such as CTDO equipment were performed. In addition, trace metal concentrations were analyzed trough ICP-MS analysis, for the same region. For biological data collection, field campaigns were performed in 2011-2012 and the picoplankton community was described by flow cytometry and taxonomical characterization with next-generation sequencing of ribosomal genes. The relation between the abiotic and biotic components was finally determined by multivariate statistical analysis. Our data show strong seasonal fluctuations in abiotic parameters such as photosynthetic active radiation and superficial sea temperature, with a clear differentiation of seasons. However, trace metal analysis allows identifying strong differentiation within the study area, dividing it into two zones based on trace metals concentration. Biological data indicate that there are no major changes in diversity but a significant fluctuation in evenness and community structure. These changes are related mainly with regional parameters, like temperature, but by analyzing the metal influence in picoplankton community structure, we identify a differential response of some plankton taxa to metal pollution. We propose that some picoeukaryotic plankton groups respond differentially to metal inputs, by changing their nutritional status and/or requirements under disturbances as a derived outcome of toxic effects and tolerance.

Keywords: Picoeukaryotes, plankton communities, trace metals, seasonal patterns

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Authors: Radik D. Aglyamov, Alexander K. Naumov, Alexey A. Shavelev, Oleg A. Morozov, Arsenij D. Shishkin, Yury P.Brodnikovsky, Alexander A.Karabutov, Alexander A. Oraevsky, Vadim V. Semashko

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The development of medical optoacoustic tomography with the using human blood as endogenic contrast agent is constrained by the lack of reliable, easy-to-use and inexpensive sources of high-power pulsed laser radiation in the spectral region of 750-900 nm [1-2]. Currently used titanium-sapphire, alexandrite lasers or optical parametric light oscillators do not provide the required and stable output characteristics, they are structurally complex, and their cost is up to half the price of diagnostic optoacoustic systems. Here we are developing the lasers based on Cr:LiCaAlF6 crystals which are free of abovementioned disadvantages and provides intensive ten’s ns-range tunable laser radiation at specific absorption bands of oxy- (~840 nm) and -deoxyhemoglobin (~757 nm) in the blood. Cr:LiCAF (с=3 at.%) crystals were grown in Kazan Federal University by the vertical directional crystallization (Bridgman technique) in graphite crucibles in a fluorinating atmosphere at argon overpressure (P=1500 hPa) [3]. The laser elements have cylinder shape with the diameter of 8 mm and 90 mm in length. The direction of the optical axis of the crystal was normal to the cylinder generatrix, which provides the π-polarized laser action correspondent to maximal stimulated emission cross-section. The flat working surfaces of the active elements were polished and parallel to each other with an error less than 10”. No any antireflection coating was applied. The Q-switched master oscillator-power amplifiers laser system (MOPA) with the dual-Xenon flashlamp pumping scheme in diffuse-reflectivity close-coupled head were realized. A specially designed laser cavity, consisting of dielectric highly reflective reflectors with a 2 m-curvature radius, a flat output mirror, a polarizer and Q-switch sell, makes it possible to operate sequentially in a circle (50 ns - laser one pulse after another) at wavelengths of 757 and 840 nm. The programmable pumping system from Tomowave Laser LLC (Russia) provided independent to each pulses (up to 250 J at 180 μs) pumping to equalize the laser radiation intensity at these wavelengths. The MOPA laser operates at 10 Hz pulse repetition rate with the output energy up to 210 mJ. Taking into account the limitations associated with physiological movements and other characteristics of patient tissues, the duration of laser pulses and their energy allows molecular and functional high-contrast imaging to depths of 5-6 cm with a spatial resolution of at least 1 mm. Highly likely the further comprehensive design of laser allows improving the output properties and realizing better spatial resolution of medical multispectral optoacoustic tomography systems.

Keywords: medical optoacoustic, endogenic contrast agent, multiwavelength tunable pulse lasers, MOPA laser system

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Authors: Miranda Lin

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This project examined the impact of a globally focused service-learning project implemented in a multicultural education course in a Midwestern university. This project facilitated critical self-reflection and build cross-cultural competence while nurturing a partnership with two schools that serve students with disabilities in Vietnam. Through a service-learning project, pre-service teachers connected via Skype with the principals/teachers at schools in Vietnam to identify and subsequently develop needed instructional materials for students with mild, moderate, and severe disabilities. Qualitative data sources include students’ intercultural competence self-reflection survey (pre-test and post-test), reflections, discussions, service project, and lesson plans. Literature Review- Global service-learning is a teaching strategy that encompasses service experiences both in the local community and abroad. Drawing on elements of global learning and international service-learning, global service-learning experiences are guided by a framework that is designed to support global learning outcomes and involve direct engagement with difference. By engaging in real-world challenges, global service-learning experiences can support the achievement of learning outcomes such as civic. Knowledge and intercultural knowledge and competence. Intercultural competence development is considered essential for cooperative and reciprocal engagement with community partners.Method- Participants (n=27*) were mostly elementary and early childhood pre-service teachers who were enrolled in a multicultural education course. All but one was female. Among the pre-service teachers, one Asian American, two Latinas, and the rest were White. Two pre-service teachers identified themselves as from the low socioeconomic families and the rest were from the middle to upper middle class.The global service-learning project was implemented in the spring of 2018. Two Vietnamese schools that served students with disabilities agreed to be the global service-learning sites. Both schools were located in an urban city.Systematic collection of data coincided with the course schedule as follows: an initial intercultural competence self-reflection survey completed in week one, guided reflections submitted in week 1, 9, and 16, written lesson plans and supporting materials for the service project submitted in week 16, and a final intercultural competence self-reflection survey completed in week 16. Significance-This global service-learning project has helped participants meet Merryfield’s goals in various degrees. They 1) learned knowledge and skills in the basics of instructional planning, 2) used a variety of instructional methods that encourage active learning, meet the different learning styles of students, and are congruent with content and educational goals, 3) gained the awareness and support of their students as individuals and as learners, 4) developed questioning techniques that build higher-level thinking skills, and 5) made progress in critically reflecting on and improving their own teaching and learning as a professional educator as a result of this project.

Keywords: global service-learning, teacher education, intercultural competence, diversity

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Authors: Maxwell Henderson, Tristan Cook, Justin Chan Jin Le, Mark Hodson, YoungJung Chang, John Novak, Daniel Padilha, Nishan Kulatilaka, Ansu Bagchi, Sanjoy Ray, John Kelly

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We present a method of using adiabatic quantum optimization (AQO) to solve a mixed integer nonlinear programming (MINLP) problem instance. The MINLP problem is a general form of a set of NP-hard optimization problems that are critical to many business applications. It requires optimizing a set of discrete and continuous variables with nonlinear and potentially nonconvex constraints. Obtaining an exact, optimal solution for MINLP problem instances of non-trivial size using classical computation methods is currently intractable. Current leading algorithms leverage heuristic and divide-and-conquer methods to determine approximate solutions. Creating more accurate and efficient algorithms is an active area of research. Quantum computing (QC) has several theoretical benefits compared to classical computing, through which QC algorithms could obtain MINLP solutions that are superior to current algorithms. AQO is a particular form of QC that could offer more near-term benefits compared to other forms of QC, as hardware development is in a more mature state and devices are currently commercially available from D-Wave Systems Inc. It is also designed for optimization problems: it uses an effect called quantum tunneling to explore all lowest points of an energy landscape where classical approaches could become stuck in local minima. Our work used a novel algorithm formulated for AQO to solve a special type of MINLP problem. The research focused on determining: 1) if the problem is possible to solve using AQO, 2) if it can be solved by current hardware, 3) what the currently achievable performance is, 4) what the performance will be on projected future hardware, and 5) when AQO is likely to provide a benefit over classical computing methods. Two different methods, integer range and 1-hot encoding, were investigated for transforming the MINLP problem instance constraints into a mathematical structure that can be embedded directly onto the current D-Wave architecture. For testing and validation a D-Wave 2X device was used, as well as QxBranch’s QxLib software library, which includes a QC simulator based on simulated annealing. Our results indicate that it is mathematically possible to formulate the MINLP problem for AQO, but that currently available hardware is unable to solve problems of useful size. Classical general-purpose simulated annealing is currently able to solve larger problem sizes, but does not scale well and such methods would likely be outperformed in the future by improved AQO hardware with higher qubit connectivity and lower temperatures. If larger AQO devices are able to show improvements that trend in this direction, commercially viable solutions to the MINLP for particular applications could be implemented on hardware projected to be available in 5-10 years. Continued investigation into optimal AQO hardware architectures and novel methods for embedding MINLP problem constraints on to those architectures is needed to realize those commercial benefits.

Keywords: adiabatic quantum optimization, mixed integer nonlinear programming, quantum computing, NP-hard

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Authors: Anupalli Roja Rani, Pavithra Dasari

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Background: Among several, breast cancer has emerged as the most common female cancer in developing countries. It is the most common cause of cancer-related deaths worldwide among women. It is a molecularly and clinically heterogeneous disease. Moreover, it is a hormone–dependent tumor in which estrogens can regulate the growth of breast cells by binding with estrogen receptors (ERs). Moreover, the use of natural products in cancer therapeutics is due to their properties of biocompatibility and less toxicity. Plants are the vast reservoirs for various bioactive compounds. Coleus barbatus (Lamiaceae) contains anticancer properties against several cancer cell lines. Method: In the present study, an attempt is being made to enrich the knowledge of the anticancer activity of pure compounds extracted from Coleus barbatus (Andrew). On human breast cancer cell lines MCF-7. Here in, we are assessing the antiproliferative activity of Coleus barbatus (Andrew) plant extracts against MCF 7 and also evaluating their toxicity in normal human mammary cell lines such as Human Mammary Epithelial Cells (HMEC). The active fraction of plant extract was further purified with the help of Flash chromatography, Medium Pressure Liquid Chromatography (MPLC) and preparative High-Performance Liquid Chromatography (HPLC). The structure of pure compounds will be elucidated by using modern spectroscopic methods like Nuclear magnetic resonance (NMR), Electrospray Ionisation Mass Spectrometry (ESI-MS) methods. Later, the growth inhibition morphological assessment of cancer cells and cell cycle analysis of purified compounds were assessed using FACS. The growth and progression of signaling molecules HER2, GRP78 was studied by secretion assay using ELISA and expression analysis by flow cytometry. Result: Cytotoxic effect against MCF-7 with IC50 values were derived from dose response curves, using six concentrations of twofold serially diluted samples, by SOFTMax Pro software (Molecular device) and respectively Ellipticine and 0.5% DMSO were used as a positive and negative control. Conclusion: The present study shows the significance of various bioactive compounds extracted from Coleus barbatus (Andrew) root material. It acts as an anti-proliferative and shows cytotoxic effects on human breast cancer cell lines MCF7. The plant extracts play an important role pharmacologically. The whole plant has been used in traditional medicine for decades and the studies done have authenticated the practice. Earlier, as described, the plant has been used in the ayurveda and homeopathy medicine. However, more clinical and pathological studies must be conducted to investigate the unexploited potential of the plant. These studies will be very useful for drug designing in the future.

Keywords: coleus barbatus, HPLC, MPLC, NMR, MCF7, flash chromatograph, ESI-MS, FACS, ELISA.

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Authors: Junali Deka

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“Pictures speak more than a thousand words”. It is the power of image which can have multiple meanings the way it is read by the viewers. This research article is an outcome of the extensive study of the phenomenon of‘selfie culture’ and dire need of self-constructed virtual identity among youths. In the recent times, there has been a revolutionary change in the concept of photography in terms of both techniques and applications. The popularity of ‘self-portraits’ mainly depend on the temporal space and time created on social networking sites like Facebook, Instagram. With reference to Stuart’s Hall encoding and decoding process, the article studies the behavior of the users who post photographs online. The photographic messages (Roland Barthes) are interpreted differently by different viewers. The notion of ‘self’, ‘self-love and practice of looking (Marita Sturken) and ways of seeing (John Berger) got new definition and dimensional together. After Oscars Night, show host Ellen DeGeneres’s selfie created the most buzz and hype in the social media. The term was judged the word of 2013, and has earned its place in the dictionary. “In November 2013, the word "selfie" was announced as being the "word of the year" by the Oxford English Dictionary. By the end of 2012, Time magazine considered selfie one of the "top 10 buzzwords" of that year; although selfies had existed long before, it was in 2012 that the term "really hit the big time an Australian origin. The present study was carried to understand the concept of ‘selfie-bug’ and the phenomenon it has created among youth (especially students) at large in developing a pseudo-image of its own. The topic was relevant and gave a platform to discuss about the cultural, psychological and sociological implications of selfie in the age of digital technology. At the first level, content analysis of the primary and secondary sources including newspapers articles and online resources was carried out followed by a small online survey conducted with the help of questionnaire to find out the student’s view on selfie and its social and psychological effects. The newspapers reports and online resources confirmed that selfie is a new trend in the digital media and it has redefined the notion of beauty and self-love. The Facebook and Instagram are the major platforms used to express one-self and creation of virtual identity. The findings clearly reflected the active participation of female students in comparison to male students. The study of the photographs of few selected respondents revealed the difference of attitude and image building among male and female users. The study underlines some basic questions about the desire of reconstruction of identity among young generation, such as - are they becoming culturally narcissist; responsible factors for cultural, social and moral changes in the society, psychological and technological effects caused by Smartphone as well, culminating into a big question mark whether the selfie is a social signifier of identity construction.

Keywords: Culture, Narcissist, Photographs, Selfie

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Authors: Rasmila Kawan, Sanjay Juvekar, Sandeep Salvi, Gufran Beig, Rainer Sauerborn

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Background: Indoor air pollution especially from the use of biomass fuels, remains a potentially large global health threat. The inefficient use of such fuels in poorly ventilated conditions results in high levels of indoor air pollution, most seriously affecting women and young children. Objectives: The main aim of this study was to measure and compare the lung function of the women exposed in the biomass fuels and LPG fuels and relate it to the indoor emission measured using a structured questionnaire, spirometer and filter based low volume samplers respectively. Methodology: This cross-sectional comparative study was conducted among the women (aged > 18 years) living in rural villages of Pune district who were not diagnosed of chronic pulmonary diseases or any other respiratory diseases and using biomass fuels or LPG for cooking for a minimum period of 5 years or more. Data collection was done from April to June 2017 in dry season. Spirometer was performed using the portable, battery-operated ultrasound Easy One spirometer (Spiro bank II, NDD Medical Technologies, Zurich, Switzerland) to determine the lung function over Forced expiratory volume. The primary outcome variable was forced expiratory volume in 1 second (FEV1). Secondary outcome was chronic obstruction pulmonary disease (post bronchodilator FEV1/ Forced Vital Capacity (FVC) < 70%) as defined by the Global Initiative for Obstructive Lung Disease. Potential confounders such as age, height, weight, smoking history, occupation, educational status were considered. Results: Preliminary results showed that the lung function of the women using Biomass fuels (FEV1/FVC = 85% ± 5.13) had comparatively reduced lung function than the LPG users (FEV1/FVC = 86.40% ± 5.32). The mean PM 2.5 mass concentration in the biomass user’s kitchen was 274.34 ± 314.90 and 85.04 ± 97.82 in the LPG user’s kitchen. Black carbon amount was found higher in the biomass users (black carbon = 46.71 ± 46.59 µg/m³) than LPG users (black carbon=11.08 ± 22.97 µg/m³). Most of the houses used separate kitchen. Almost all the houses that used the clean fuel like LPG had minimum amount of the particulate matter 2.5 which might be due to the background pollution and cross ventilation from the houses using biomass fuels. Conclusions: Therefore, there is an urgent need to adopt various strategies to improve indoor air quality. There is a lacking of current state of climate active pollutants emission from different stove designs and identify major deficiencies that need to be tackled. Moreover, the advancement in research tools, measuring technique in particular, is critical for researchers in developing countries to improve their capability to study the emissions for addressing the growing climate change and public health concerns.

Keywords: black carbon, biomass fuels, indoor air pollution, lung function, particulate matter

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Authors: Rose Branch-Allen, John Jayachandran

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Background and purpose: Canada is a pluralistic, multicultural society with an ethno-cultural composition that has been shaped over time by immigrants and their descendants. Although Canada welcomes these immigrants, many will endure hardship and assimilation difficulties. Despite these life hurdles, surveys consistently disclose high life satisfaction for all Canadians. Most research studies on Life Satisfaction/ Subjective Wellbeing (SWB) have focused on one main determinant and a variety of social demographic variables to delineate the determinants of life satisfaction. However, very few research studies examine life satisfaction from a holistic approach. In addition, we need to understand the causal pathways leading to life satisfaction, and develop theories that explain why certain variables differentially influence the different components of SWB. The aim this study was to utilize a holistic approach to construct a causal model and identify major determinants of life satisfaction. Data and measures: This study utilized data from the General Social Survey, with a sample size of 19, 597. The exogenous concepts included age, gender, marital status, household size, socioeconomic status, ethnicity, location, immigration status, religiosity, and neighborhood. The intervening concepts included health, social contact, leisure, enjoyment, work-family balance, quality time, domestic labor, and sense of belonging. The endogenous concept life satisfaction was measured by multiple indicators (Cronbach’s alpha = .83). Analysis: Several multiple regression models were run sequentially to estimate path coefficients for the causal model. Results: Overall, above average satisfaction with life was reported for respondents with specific socio-economic, demographic and lifestyle characteristics. With regard to exogenous factors, respondents who were female, younger, married, from high socioeconomic status background, born in Canada, very religious, and demonstrated high level of neighborhood interaction had greater satisfaction with life. Similarly, intervening concepts suggested respondents had greater life satisfaction if they had better health, more social contact, less time on passive leisure activities and more time on active leisure activities, more time with family and friends, more enjoyment with volunteer activities, less time on domestic labor and a greater sense of belonging to the community. Conclusions and Implications: Our results suggest that a holistic approach is necessary for establishing determinants of life satisfaction, and that life satisfaction is not merely comprised of positive or negative affect rather understanding the causal process of life satisfaction. Even though, most of our findings are consistent with previous studies, a significant number of causal connections contradict some of the findings in literature today. We have provided possible explanation for these anomalies researchers encounter in studying life satisfaction and policy implications.

Keywords: causal model, holistic approach, life satisfaction, socio-demographic variables, subjective well-being

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Authors: Emma Castiglioni, Nigel Scrutton, Derren Heyes, Alistair Fielding

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By using light as trigger, it is possible to study many biological processes, such as the activity of genes, proteins, and other molecules, with precise spatiotemporal control. Caged compounds, where biologically active molecules are generated from an inert precursor upon laser photolysis, offer the potential to initiate such biological reactions with high temporal resolution. As light acts as the trigger for cleaving the protecting group, the ‘caging’ technique provides a number of advantages as it can be intracellular, rapid and controlled in a quantitative manner. We are developing caging strategies to study the catalytic cycle of a number of enzyme systems, such as nitric oxide synthase and ethanolamine ammonia lyase. These include the use of caged substrates, caged electrons and the possibility of caging the enzyme itself. In addition, we are developing a novel freeze-quench instrument to study these reactions, which combines rapid mixing and flashing capabilities. Reaction intermediates will be trapped at low temperatures and will be analysed by using electron paramagnetic resonance (EPR) spectroscopy to identify the involvement of any radical species during catalysis. EPR techniques typically require relatively long measurement times and very often, low temperatures to fully characterise these short-lived species. Therefore, common rapid mixing techniques, such as stopped-flow or quench-flow are not directly suitable. However, the combination of rapid freeze-quench (RFQ) followed by EPR analysis provides the ideal approach to kinetically trap and spectroscopically characterise these transient radical species. In a typical RFQ experiment, two reagent solutions are delivered to the mixer via two syringes driven by a pneumatic actuator or stepper motor. The new mixed solution is then sprayed into a cryogenic liquid or surface, and the frozen sample is then collected and packed into an EPR tube for analysis. The earliest RFQ instrument consisted of a hydraulic ram unit as a drive unit with direct spraying of the sample into a cryogenic liquid (nitrogen, isopentane or petroleum). Improvements to the RFQ technique have arisen from the design of new mixers in order to reduce both the volume and the mixing time. In addition, the cryogenic isopentane bath has been coupled to a filtering system or replaced by spraying the solution onto a surface that is frozen via thermal conductivity with a cryogenic liquid. In our work, we are developing a novel RFQ instrument which combines the freeze-quench technology with flashing capabilities to enable the studies of both thermally-activated and light-activated biological reactions. This instrument also uses a new rotating plate design based on magnetic couplings and removes the need for mechanical motorised rotation, which can otherwise be problematic at cryogenic temperatures.

Keywords: caged compounds, freeze-quench apparatus, photolysis, radicals

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Authors: Frea Van Steenweghen, Lander Hollevoet, Johan A. Martens

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Compared to other hydrogen (H₂) carriers, ammonia (NH₃) is one of the most promising carriers as it contains 17.6 wt% hydrogen. It is easily liquefied at ≈ 9–10 bar pressure at ambient temperature. More importantly, NH₃ is a carbon-free hydrogen carrier with no CO₂ emission at final decomposition. Ammonia has a well-defined regulatory framework and a good track record regarding safety concerns. Furthermore, the industry already has an existing transport infrastructure consisting of pipelines, tank trucks and shipping technology, as ammonia has been manufactured and distributed around the world for over a century. While NH₃ synthesis and transportation technological solutions are at hand, a missing link in the hydrogen delivery scheme from ammonia is an energy-lean and efficient technology for cracking ammonia into H₂ and N₂. The most explored option for ammonia decomposition is thermo-catalytic cracking which is, by itself, the most energy-efficient approach compared to other technologies, such as plasma and electrolysis, as it is the most energy-lean and robust option. The decomposition reaction is favoured only at high temperatures (> 300°C) and low pressures (1 bar) as the thermocatalytic ammonia cracking process is faced with thermodynamic limitations. At 350°C, the thermodynamic equilibrium at 1 bar pressure limits the conversion to 99%. Gaining additional conversion up to e.g. 99.9% necessitates heating to ca. 530°C. However, reaching thermodynamic equilibrium is infeasible as a sufficient driving force is needed, requiring even higher temperatures. Limiting the conversion below the equilibrium composition is a more economical option. Thermocatalytic ammonia cracking is documented in scientific literature. Among the investigated metal catalysts (Ru, Co, Ni, Fe, …), ruthenium is known to be most active for ammonia decomposition with an onset of cracking activity around 350°C. For establishing > 99% conversion reaction, temperatures close to 600°C are required. Such high temperatures are likely to reduce the round-trip efficiency but also the catalyst lifetime because of the sintering of the supported metal phase. In this research, the first focus was on catalyst bed design, avoiding diffusion limitation. Experiments in our packed bed tubular reactor set-up showed that extragranular diffusion limitations occur at low concentrations of NH₃ when reaching high conversion, a phenomenon often overlooked in experimental work. A second focus was thermocatalyst development for ammonia cracking, avoiding the use of noble metals. To this aim, candidate metals and mixtures were deposited on a range of supports. Sintering resistance at high temperatures and the basicity of the support were found to be crucial catalyst properties. The catalytic activity was promoted by adding alkaline and alkaline earth metals. A third focus was studying the optimum process configuration by process simulations. A trade-off between conversion and favorable operational conditions (i.e. low pressure and high temperature) may lead to different process configurations, each with its own pros and cons. For example, high-pressure cracking would eliminate the need for post-compression but is detrimental for the thermodynamic equilibrium, leading to an optimum in cracking pressure in terms of energy cost.

Keywords: ammonia cracking, catalyst research, kinetics, process simulation, thermodynamic equilibrium

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Authors: M. Grandcolas, N. Rival, H. Bu, S. Jahren, R. Schmid, H. Johnsen

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The concept of an autonomic self-healing material, where initiation of repair is integrated to the material, is now being considered for engineering applications and is a hot topic in the literature. Among several concepts/techniques, two are most interesting: i) Capsules: Integration of microcapsules in or at the surface of coatings or fibre-like structures has recently gained much attention. Upon damage-induced cracking, the microcapsules are broken by the propagating crack fronts resulting in a release of an active chemical (healing agent) by capillary action, subsequently repairing and avoiding further crack growth. ii) Self-healing polymers: Interestingly, the introduction of dynamic covalent bonds into polymer networks has also recently been used as a powerful approach towards the design of various intrinsically self-healing polymer systems. The idea behind this is to reconnect the chemical crosslinks which are broken when a material fractures, restoring the integrity of the material and thereby prolonging its lifetime. We propose here to integrate both self-healing concepts (capsules, self-healing polymers) in electrospun fibres and coatings. Different capsule preparation approaches have been investigated in SINTEF. The most advanced method to produce capsules is based on emulsification to create a water-in-oil emulsion before polymerisation. The healing agent is a polyurethane-based dispersion that was encapsulated in shell materials consisting of urea-benzaldehyde resins. Results showed the successful preparation of microcapsules and release of the agent when capsules break. Since capsules are produced in water-in-oil systems we mainly investigated organic solvent based coatings while a major challenge resides in the incorporation of capsules into water-based coatings. We also focused on developing more robust microcapsules to prevent premature rupture of the capsules. The capsules have been characterized in terms of size, and encapsulation and release might be visualized by incorporating fluorescent dyes and examine the capsules by microscopy techniques. Alternatively, electrospinning is an innovative technique that has attracted enormous attention due to unique properties of the produced nano-to-micro fibers, ease of fabrication and functionalization, and versatility in controlling parameters. Especially roll-to-roll electrospinning is a unique method which has been used in industry to produce nanofibers continuously. Electrospun nanofibers can usually reach a diameter down to 100 nm, depending on the polymer used, which is of interest for the concept with self-healing polymer systems. In this work, we proved the feasibility of fabrication of POSS-based (POSS: polyhedral oligomeric silsesquioxanes, tradename FunzioNano™) nanofibers via electrospinning. Two different formulations based on aqueous or organic solvents have shown nanofibres with a diameter between 200 – 450nm with low defects. The addition of FunzioNano™ in the polymer blend also showed enhanced properties in term of wettability, promising for e.g. membrane technology. The self-healing polymer systems developed are here POSS-based materials synthesized to develop dynamic soft brushes.

Keywords: capsules, coatings, electrospinning, fibers

Procedia PDF Downloads 240

Authors: M. Pienaar, J. W. Odendaal, J. C. Smit, J. Joubert

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Simulations are commonly used to predict the bistatic radar cross section (RCS) of military targets since characterization measurements can be expensive and time consuming. It is thus important to accurately predict the bistatic RCS of targets. Computational electromagnetic (CEM) methods can be used for bistatic RCS prediction. CEM methods are divided into full-wave and asymptotic methods. Full-wave methods are numerical approximations to the exact solution of Maxwell’s equations. These methods are very accurate but are computationally very intensive and time consuming. Asymptotic techniques make simplifying assumptions in solving Maxwell's equations and are thus less accurate but require less computational resources and time. Asymptotic techniques can thus be very valuable for the prediction of bistatic RCS of electrically large targets, due to the decreased computational requirements. This study extends previous work by validating the accuracy of asymptotic techniques to predict bistatic RCS through comparison with full-wave simulations as well as measurements. Validation is done with canonical structures as well as complex realistic aircraft models instead of only looking at a complex slicy structure. The slicy structure is a combination of canonical structures, including cylinders, corner reflectors and cubes. Validation is done over large bistatic angles and at different polarizations. Bistatic RCS measurements were conducted in a compact range, at the University of Pretoria, South Africa. The measurements were performed at different polarizations from 2 GHz to 6 GHz. Fixed bistatic angles of β = 30.8°, 45° and 90° were used. The measurements were calibrated with an active calibration target. The EM simulation tool FEKO was used to generate simulated results. The full-wave multi-level fast multipole method (MLFMM) simulated results together with the measured data were used as reference for validation. The accuracy of physical optics (PO) and geometrical optics (GO) was investigated. Differences relating to amplitude, lobing structure and null positions were observed between the asymptotic, full-wave and measured data. PO and GO were more accurate at angles close to the specular scattering directions and the accuracy seemed to decrease as the bistatic angle increased. At large bistatic angles PO did not perform well due to the shadow regions not being treated appropriately. PO also did not perform well for canonical structures where multi-bounce was the main scattering mechanism. PO and GO do not account for diffraction but these inaccuracies tended to decrease as the electrical size of objects increased. It was evident that both asymptotic techniques do not properly account for bistatic structural shadowing. Specular scattering was calculated accurately even if targets did not meet the electrically large criteria. It was evident that the bistatic RCS prediction performance of PO and GO depends on incident angle, frequency, target shape and observation angle. The improved computational efficiency of the asymptotic solvers yields a major advantage over full-wave solvers and measurements; however, there is still much room for improvement of the accuracy of these asymptotic techniques.

Keywords: asymptotic techniques, bistatic RCS, geometrical optics, physical optics

Procedia PDF Downloads 232

Authors: Meilan Jin, Mary Jane Moran

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This study aims to introduce the framework of culturally responsive home visiting (CRHV) to head start teacher professional sessions in the Southeastern of the US and investigate its influence on the evolving beliefs of teachers about their roles and relationships with families in-home visits. The framework orients teachers to an effective way of taking on the role of learner to listen for spoken and unspoken needs and look for family strengths. In addition, it challenges the deficit model that is grounded on 'cultural deprivation,' and it stresses the value of family cultures and advocates equal, collaborative parent-teacher relationships. The home visit reflection papers and focus group transcriptions of eight teachers have been collected since 2010 throughout a five-year longitudinal collaboration with them. Reflection papers were written by the teachers before and after introducing the CRHV framework, including the details of visit purposes and actions and their plans for later home visits. Particularly, the CRHV framework guided the teachers to listen and look for information about family-living environments; parent-child interactions; child-rearing practices; and parental beliefs, values, and needs. Two focus groups were organized in 2014 by asking the teachers to read their written reflection papers and then discussing their shared beliefs and experiences of home visits in recent years. The average length of the discussions was one hour, and the discussions were audio-recorded and transcribed verbatim. Moreover, the data were analyzed using constant comparative analysis, and the analysis was verified through (a) the uses of multiple data sources, (b) the involvement of multiple researchers, (c) coding checks, and (d) the provisions of the thick descriptions of the findings. The study findings corroborate that the teachers become to reposition themselves as 'knowledge seekers' through reorienting their cynosure toward 'setting stones' to learn, grow, and change rather than framing their home visits. The teachers also continually engage in careful listening, observing, questioning, and dialoguing, and these actions reflect their care toward parents. The value of teamwork with parents is advocated, and the teachers recognize that when parents feel empowered, they are active and committed to doing more for their children, which can further advantage proactive long-term parent-teacher collaborations. The study findings also validate that the framework is influential for educators to provide the experiences of home visiting that is culturally responsive and to share collaborative relationships with caregivers. The long-term impact of the framework further implies that teachers continue to put themselves in the position of evolving, including beliefs and actions, to better work with children and families who are culturally, ethnically, and linguistically different from them. This framework can be applicable to educators and professionals who are looking for avenues to bridge the relationship between home and school and parents and teachers.

Keywords: culturally responsive home visit, early childhood education, parent–teacher collaboration, teacher professional development

Procedia PDF Downloads 79

Authors: Ben Morrow, Patrick O'Hara, Natalie Ban, Tunai Marques, Molly Fraser, Christopher Bone

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Relative to commercial fisheries, recreational fisheries are often poorly understood and pose various challenges for monitoring frameworks. In British Columbia (BC), Canada, Pacific salmon are heavily targeted by recreational fishers while also being a key source of nutrient flow and crucial prey for a variety of marine and terrestrial fauna, including endangered Southern Resident killer whales (Orcinus orca). Although commercial fisheries were historically responsible for the majority of salmon retention, recreational fishing now comprises both greater effort and retention. The current monitoring scheme for recreational salmon fisheries involves aerial-roving creel surveys. However, this method has been identified as costly and having low predictive power as it is often limited to sampling fragments of fluid and temporally dynamic fisheries. This study used imagery from two shore-based autonomous cameras in a highly active recreational fishery around Sooke, BC, and evaluated their efficacy in supplementing existing aerial-roving surveys for monitoring a recreational salmon fishery. This study involved continuous monitoring and high temporal resolution (over one million images analyzed in a single fishing season), using a deep learning-based vessel detection algorithm and a custom image annotation tool to efficiently thin datasets. This allowed for the quantification of peak-season effort from a busy harbour, species-specific retention estimates, high levels of detected fishing events at a nearby popular fishing location, as well as the proportion of the fishery management area represented by cameras. Then, this study demonstrated how it could substantially enhance the temporal resolution of a fishery through diel activity pattern analyses, scaled monthly to visualize clusters of activity. This work also highlighted considerable off-season fishing detection, currently unaccounted for in the existing monitoring framework. These results demonstrate several distinct applications of autonomous cameras for providing enhanced detail currently unavailable in the current monitoring framework, each of which has important considerations for the managerial allocation of resources. Further, the approach and methodology can benefit other studies that apply shore-based camera monitoring, supplement aerial-roving creel surveys to improve fine-scale temporal understanding, inform the optimal timing of creel surveys, and improve the predictive power of recreational stock assessments to preserve important and endangered fish species.

Keywords: cameras, monitoring, recreational fishing, stock assessment

Procedia PDF Downloads 93

Authors: Oleg Reva, Ilya Korotetskiy, Marina Lankina, Murat Kulmanov, Aleksandr Ilin

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Molecular docking approaches are widely used for design of new antibiotics and modeling of antibacterial activities of numerous ligands which bind specifically to active centers of indispensable enzymes and/or key signaling proteins of pathogens. Widespread drug resistance among pathogenic microorganisms calls for development of new antibiotics specifically targeting important metabolic and information pathways. A generally recognized problem is that almost all molecular targets have been identified already and it is getting more and more difficult to design innovative antibacterial compounds to combat the drug resistance. A promising way to overcome the drug resistance problem is an induction of reversion of drug resistance by supplementary medicines to improve the efficacy of the conventional antibiotics. In contrast to well established computer-based drug design, modeling of drug resistance reversion still is in its infancy. In this work, we proposed an approach to identification of compensatory genetic variants reducing the fitness cost associated with the acquisition of drug resistance by pathogenic bacteria. The approach was based on an analysis of the population genetic of Mycobacterium tuberculosis and on results of experimental modeling of the drug resistance reversion induced by a new anti-tuberculosis drug FS-1. The latter drug is an iodine-containing nanomolecular complex that passed clinical trials and was admitted as a new medicine against MDR-TB in Kazakhstan. Isolates of M. tuberculosis obtained on different stages of the clinical trials and also from laboratory animals infected with MDR-TB strain were characterized by antibiotic resistance, and their genomes were sequenced by the paired-end Illumina HiSeq 2000 technology. A steady increase in sensitivity to conventional anti-tuberculosis antibiotics in series of isolated treated with FS-1 was registered despite the fact that the canonical drug resistance mutations identified in the genomes of these isolates remained intact. It was hypothesized that the drug resistance phenotype in M. tuberculosis requires an adjustment of activities of many genes to compensate the fitness cost of the drug resistance mutations. FS-1 cased an aggravation of the fitness cost and removal of the drug-resistant variants of M. tuberculosis from the population. This process caused a significant increase in genetic heterogeneity of the Mtb population that was not observed in the positive and negative controls (infected laboratory animals left untreated and treated solely with the antibiotics). A large-scale search for linkage disequilibrium associations between the drug resistance mutations and genetic variants in other genomic loci allowed identification of target proteins, which could be influenced by supplementary drugs to increase the fitness cost of the drug resistance and deprive the drug-resistant bacterial variants of their competitiveness in the population. The approach will be used to improve the efficacy of FS-1 and also for computer-based design of new drugs to combat drug-resistant infections.

Keywords: complete genome sequencing, computational modeling, drug resistance reversion, Mycobacterium tuberculosis

Procedia PDF Downloads 241

Authors: Vanessa G. P. Severino, Eliangela Cristina Candida Costa, Nubia Alves Mariano Teixeira Pires Gomides, Lucilia Kato, Afif Felix Monteiro, Maria Anita Lemos Vasconcelos Ambrosio, Carlos Henrique Gomes Martins

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One of the biomes present in Brazil is known as Cerrado, which is a vast tropical savanna ecoregion, particularly in the states of Goiás, Mato Grosso do Sul, Mato Grosso, Tocantins and Minas Gerais. Many species of plants are characterized as endemic and they have therapeutic value for a large part of the population, especially to the rural communities. Given that, the southeastern region of the state of Goiás contains about 21 rural communities, which present a form of organization based on the use of natural resources available. One of these rural communities is named of Coqueiros, where the knowledge about the medicinal plants was very important to this research. Thus, this study focuses on the ethnobotanical survey of this community on the use of Kielmeyera coriacea to treat diseases. From the 37 members interviewed, 76% indicated this species for the treatment of intestinal infection, leukemia, anemia, gastritis, gum pain, toothache, cavity, arthritis, arthrosis, healing, vermifuge, rheumatism, antibiotic, skin problems, mycoses and all kinds of infections. The medicinal properties attributed during the interviews were framed in the body system (disease categories), adapted from ICD 10; thus, 20 indications of use were obtained, among five body systems. Therefore, the root of this species was select to chemical and biological (antioxidant and antimicrobial) studies. From the liquid-liquid extraction of ethanolic extract of root (EER), the hexane (FH), ethyl acetate (FAE), and hydro alcoholic (FHA) fractions were obtained. The chemical profile study of these fractions was performed by LC-MS, identifying major compounds such as δ-tocotrienol, prenylated acylphoroglucinol, 2-hydroxy-1-methoxyxanthone and quercitrin. EER, FH, FAE and FHA were submitted to biological tests. FHA presented the best antioxidant action (EC50 201.53 μg mL-1). EER inhibited the bacterial growth of Streptococcus pyogenes and Pseudomonas aeruginosa, microorganisms associated with rheumatism, at Minimum Inhibitory Concentration (MIC) of 6.25 μg mL-1. In addition, the FH-10 subfraction, obtained from FH fractionation, presented MIC of 1.56 μg mL-1 against S. pneumoniae; EER also inhibited the fungus Candida glabrata (MIC 7.81 μg mL- 1). The FAE-4.7.3 fraction, from the fractionation of FAE, presented MIC of 200 μg mL-1 against Lactobacillus casei, which is one of the causes of caries and oral infections. By the correlation of the chemical and biological data, it is possible to note that the FAE-4.7.3 and FH-10 are constituted 4-hydroxy-2,3-methylenedioxy xanthone, 3-hydroxy-1,2-dimethoxy xanthone, lupeol, prenylated acylphoroglucinol and quercitrin, which could be associated with the biological potential found. Therefore, this study provides an important basis for further investigations regarding the compounds present in the active fractions of K. coriacea, which will permit the establishment of a correlation between ethnobotanical survey and bioactivity.

Keywords: biological activity, ethnobotanical survey, Kielmeyera coriacea Mart., LC-MS profile

Procedia PDF Downloads 115

Authors: H. M. Ross-McAlpine

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Much of what is written on diplomacy, globalization and the global economy addresses the changing nature of relationships between major powers. While the most dramatic and influential changes have resulted from these developing relationships the world is not, on deeper inspection, governed neatly by major powers. Due to advances in technology, the shifting balance of power and a changing geopolitical order, small states have the ability to exercise a greater influence than ever before. Increasingly interdependent and ever complex, our world is too delicate to be handled by a mighty few. The pressure of global change requires small states to adapt their diplomatic practices and diversify their strategic alliances and relationships. The nature and practice of diplomacy must be re-evaluated in light of the pressures resulting from globalization. This research examines: how small states can best secure their foreign policy objectives? Small state theory is used as a foundation for exploring the case study of New Zealand. The research draws on secondary sources to evaluate the existing theory in relation to modern practices of diplomacy. As New Zealand lacks the required economic and military power to play an active, influential role in international affairs what strategies are used to exert influence? Furthermore, New Zealand lies in a remote corner of the Pacific and is geographically isolated from its nearest neighbors how does this affect security and trade priorities? The findings note a significant shift since the 1970’s in New Zealand’s diplomatic relations. This shift is arguably a direct result of globalization, regionalism and a growing independence from the traditional bi-lateral relationships. The need to source predictable trade, investment and technology are an essential driving force for New Zealand’s diplomatic relations. A lack of hard power aligns New Zealand’s prosperity with a secure, rules-based international system that increases the likelihood of a stable and secure global order. New Zealand’s diplomacy and prosperity has been intrinsically reliant on its reputation. A vital component of New Zealand’s diplomacy is preserving a reputation for integrity and global responsibility. It is the use of this soft power that facilitates the influence that New Zealand enjoys on the world stage. To weave a comprehensive network of successful diplomatic relationships, New Zealand must maintain a reputation of international credibility. Globalization has substantially influenced the practice of diplomacy for New Zealand. The current world order places economic and military might in the hands of a few, subsequently requiring smaller states to use other means for securing their interests. There are clear strategies evident in New Zealand’s diplomacy practice that draw attention to how other smaller states might best secure their foreign policy objectives. While these findings are limited, as with all case study research, there is value in applying the findings to other small states struggling to secure their interests in the wake of rapid globalization.

Keywords: diplomacy, foreign policy, globalisation, small state

Procedia PDF Downloads 364

Authors: Boudjelal Amel

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Arisarum vulgare, a member of the Araceae family, is an herbaceous perennial widely distributed in the Mediterranean region. A. vulgare is recognized for its medicinal properties and holds significant traditional importance in Algeria for the treatment of various human ailments, including pain, infections, inflammation, digestive disorders, skin problems, eczema, cancer, wounds, burns and gynecological diseases. Despite its extensive traditional use, scientific exploration of A. vulgare remains limited. The study aims to investigate for the first time the therapeutic potential of A. vulgare ethanolic extract obtained by ultrasound-assisted extraction. The chemical composition of the extract was determined by LC-MS/MS analysis. For in vitro phytopharmacological evaluation, several assays, including DPPH, ABTS, FRAP and reducing power, were employed to evaluate the antioxidant activity. The antibacterial activity was assessed againt Escherichia coli, Salmonella typhimurium, Staphylococus aureus, Enterococcus feacium by disk diffusion and microdilution methods. The possible inhibitory activity of ethanolic extract was analyzed against the cholinesterases enzymes (AChE and BChE). The DNA protection activity of A. vulgare ethanolic extract was estimated using the agarose gel electrophoresis method. The capacities of the extract to protect plasmid DNA (pBR322) from the oxidizing effects of H2O2 and UV treatment were evaluated by their DNA-breaking forms. The in vivo wound healing potential of a traditional ointment containing 5% of A. vulgare ethanolic extract was also investigated. The LC-MS/MS profiling of the extract revealed the presence of various bioactive compounds, including naringenin, chlorogenic, vanillic, cafeic, coumaric acids, trans-cinnamic and trans ferrulic acids. The plant extract presented considerable antioxidant potential, being the most active for Reducing power (0,07326±0.001 mg/ml) and DPPH (0.14±0.004 mg/ml). The extract showed the highest inhibition zone diameter against Enterococcus feacium (36±0.1 mm). The ethanolic extract of A. vulgare suppressed the growth of Staphylococus aureus, Escherichia coli and Salmonella typhimurium according to the MIC values. The extract of the plant significantly inhibited both AChE and BChE enzymes. DNA protection activity of the A. vulgare extract was determined as 90.41% for form I and 51.92% for form II. The in vivo experiments showed that 5% ethanolic extract ointment accelerated the wound healing process. The topical application of the traditional formulation enhanced wound closure (95,36±0,6 %) and improved histological parameters in the treated group compared to the control groups. The promising biological properties of Arisarum vulgare revealed that the plant could be appraised as a potential origin of bioactive molecules having multifunctional medicinal uses.

Keywords: arisarum vulgare, LC-MS/MS, antioxidant activity, antimicrobial activity, cholinesterases enzymes inhibition, dna-damage activity, in vivo wound healing

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Authors: Tarun Kumar Dalakoti, Debajyoti Majumder, Aditya Prasad Das, Samir Chandra Saxena

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India’s ambitious target to achieve a 50 percent share of energy from non-fossil fuels and the 500-gigawatt (GW) renewable energy capacity before the deadline of 2030, coupled with the global pursuit of sustainable development, will compel the nation to embark on a rapid clean energy transition. As a result, electricity market reforms will emerge as critical policy instruments to facilitate this transition and achieve ambitious environmental targets. This paper will present a comprehensive analysis of the various electricity market reforms to be introduced in the Indian Electricity sector to facilitate the integration of clean energy sources and will assess their impact on the overall energy landscape. The first section of this paper will delve into the policy mechanisms to be introduced by the Government of India and the Central Electricity Regulatory Commission to promote clean energy deployment. These mechanisms include extensive provisions for the integration of renewables in the Indian Electricity Grid Code, 2023. The section will also cover the projection of RE Generation as highlighted in the National Electricity Plan, 2023. It will discuss the introduction of Green Energy Market segments, the waiver of Inter-State Transmission System (ISTS) charges for inter-state sale of solar and wind power, the notification of Promoting Renewable Energy through Green Energy Open Access Rules, and the bundling of conventional generating stations with renewable energy sources. The second section will evaluate the tangible impact of these electricity market reforms. By drawing on empirical studies and real-world case examples, the paper will assess the penetration rate of renewable energy sources in India’s electricity markets, the decline of conventional fuel-based generation, and the consequent reduction in carbon emissions. Furthermore, it will explore the influence of these reforms on electricity prices, the impact on various market segments due to the introduction of green contracts, and grid stability. The paper will also discuss the operational challenges to be faced due to the surge of RE Generation sources as a result of the implementation of the above-mentioned electricity market reforms, including grid integration issues, intermittency concerns with renewable energy sources, and the need for increasing grid resilience for future high RE in generation mix scenarios. In conclusion, this paper will emphasize that electricity market reforms will be pivotal in accelerating the global transition towards clean energy systems. It will underscore the importance of a holistic approach that combines effective policy design, robust regulatory frameworks, and active participation from market actors. Through a comprehensive examination of the impact of these reforms, the paper will shed light on the significance of India’s sustained commitment to a cleaner, more sustainable energy future.

Keywords: renewables, Indian electricity grid code, national electricity plan, green energy market

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Authors: Purushottam P. Khatiwada, Bikash Paudel, Ram B. Rana, Parshuram Biswakarma, Roshan Pudasaini

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Nepal has been putting its efforts into securing food and nutrition security for its citizens adopting different models and approaches. Home Garden approach, that integrates vegetables, fruits, small livestock, poultry along with other components like fish, honeybee, mushroom, spices for the promotion of nutritional security of resource-poor and disadvantaged groups was implemented during March 2009 to July 2013 spreading over 16 districts of Nepal covering 115 farmers groups, directly working with 3500 households. Sustained long-term impact of development interventions targeted to the resource-poor and disadvantaged groups has been a recurrent issue for donors, policymakers and practitioners alike. Considering the issue, a post-project evaluation was carried out in a selected project group (Dangibari of Jhapa) after four years of project completion in 2017 in order to evaluate the impact and understand the factors associated with its success. Qualitative information was collected through focus group discussion with group members and associated local institutions. For quantitative information, a quick survey was carried out to the same group members only selecting few indicators. The results are compared with the data obtained from the baseline study conducted by the project in March 2009. The impact of project intervention was evident as compared to the benchmarks established during the baseline, even after four years of project completion. The area under home garden is increased to 729 m² from 386 m² and average food self-sufficiency months increased to 10.22 from 8.11. Seven to eleven fruit species are maintained in the home gardens. An average number of vegetable species grown increased to 15.85 from 9.86. It has resulted in an increase in vegetables self-sufficient month to 8.74 from 4.74 and a huge increase in cash income NPR 6142.8 (USD 59.6) from NPR 385.7 (USD 3.9) from the sale of surplus vegetables. Coaching and mentoring including nutrition sensitization by the project staff at the beginning, inputs and technical support during the project implementation phase and projects effort on the institutional building of disadvantaged farmers were the key drivers of home garden sustainability and expansion. Specifically, package of home garden management trainings provided by the project staff, availability of group funds for buying inputs even after the project, uniting home garden group members in a cooperative, resource leveraging by local institutions through group lobbying, farmers innovations for maintaining home garden diversity and continuous backstopping support by few active members as local resource persons to other members are some additional factors contributing to sustain and/or improve the home garden status by the resource-poor and disadvantaged group.

Keywords: food-based nutrition, home garden, resource-poor and disadvantaged group, sustained impact

Procedia PDF Downloads 118

Authors: F. Infante, W. Kaal, S. Perfetto, S. Herold

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In the last decade the demand of wireless sensors and low-power electric devices for condition monitoring in mechanical structures has been strongly increased. Networks of wireless sensors can potentially be applied in a huge variety of applications. Due to the reduction of both size and power consumption of the electric components and the increasing complexity of mechanical systems, the interest of creating dense nodes sensor networks has become very salient. Nevertheless, with the development of large sensor networks with numerous nodes, the critical problem of powering them is drawing more and more attention. Batteries are not a valid alternative for consideration regarding lifetime, size and effort in replacing them. Between possible alternative solutions for durable power sources useable in mechanical components, vibrations represent a suitable source for the amount of power required to feed a wireless sensor network. For this purpose, energy harvesting from structural vibrations has received much attention in the past few years. Suitable vibrations can be found in numerous mechanical environments including automotive moving structures, household applications, but also civil engineering structures like buildings and bridges. Similarly, a dynamic vibration absorber (DVA) is one of the most used devices to mitigate unwanted vibration of structures. This device is used to transfer the primary structural vibration to the auxiliary system. Thus, the related energy is effectively localized in the secondary less sensitive structure. Then, the additional benefit of harvesting part of the energy can be obtained by implementing dedicated components. This paper describes the design process of an energy harvesting tuned vibration absorber (EHTVA) for rotating systems using piezoelectric elements. The energy of the vibration is converted into electricity rather than dissipated. The device proposed is indeed designed to mitigate torsional vibrations as with a conventional rotational TVA, while harvesting energy as a power source for immediate use or storage. The resultant rotational multi degree of freedom (MDOF) system is initially reduced in an equivalent single degree of freedom (SDOF) system. The Den Hartog’s theory is used for evaluating the optimal mechanical parameters of the initial DVA for the SDOF systems defined. The performance of the TVA is operationally assessed and the vibration reduction at the original resonance frequency is measured. Then, the design is modified for the integration of active piezoelectric patches without detuning the TVA. In order to estimate the real power generated, a complex storage circuit is implemented. A DC-DC step-down converter is connected to the device through a rectifier to return a fixed output voltage. Introducing a big capacitor, the energy stored is measured at different frequencies. Finally, the electromechanical prototype is tested and validated achieving simultaneously reduction and harvesting functions.

Keywords: energy harvesting, piezoelectricity, torsional vibration, vibration absorber

Procedia PDF Downloads 118

Authors: Rahaba Marima, Clement Penny

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The Health-related quality of life (HRQoL) for HIV positive patients has improved since the introduction of the highly active antiretroviral treatment (HAART). However, in the present HAART era, HIV co-morbidities such as lung cancer, a non-AIDS (NAIDS) defining cancer have been documented to be on the rise. Under normal physiological conditions, cells grow, repair and proliferate through the cell-cycle as cellular homeostasis is important in the maintenance and proper regulation of tissues and organs. Contrarily, the deregulation of the cell-cycle is a hallmark of cancer, including lung cancer. The association between lung cancer and the use of HAART components such as Efavirenz (EFV) is poorly understood. This study aimed at elucidating the effects of EFV on the cell-cycle genes’ expression in lung cancer. For this purpose, the human cell-cycle gene array composed of 84 genes was evaluated on both normal lung fibroblasts (MRC-5) cells and adenocarcinoma (A549) lung cells, in response to 13µM EFV or 0.01% vehicle. The ±2 up or down fold change was used as a basis of target selection, with p < 0.05. Additionally, RT-qPCR was done to validate the gene array results. Next, In-silico bio-informatics tools, Search Tool for the Retrieval of Interacting Genes/Proteins (STRING), Reactome, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway and Ingenuity Pathway Analysis (IPA) were used for gene/gene interaction studies as well as to map the molecular and biological pathways influenced by the identified targets. Interestingly, the DNA damage response (DDR) pathway genes such as p53, Ataxia telangiectasia mutated and Rad3 related (ATR), Growth arrest and DNA damage inducible alpha (GADD45A), HUS1 checkpoint homolog (HUS1) and Role of radiation (RAD) genes were shown to be upregulated following EFV treatment, as revealed by STRING analysis. Additionally, functional enrichment analysis by the KEGG pathway revealed that most of the differentially expressed gene targets function at the cell-cycle checkpoint such as p21, Aurora kinase B (AURKB) and Mitotic Arrest Deficient-Like 2 (MAD2L2). Core analysis by IPA revealed that p53 downstream targets such as survivin, Bcl2, and cyclin/cyclin dependent kinases (CDKs) complexes are down-regulated, following exposure to EFV. Furthermore, Reactome analysis showed a significant increase in cellular response to stress genes, DNA repair genes, and apoptosis genes, as observed in both normal and cancerous cells. These findings implicate the genotoxic effects of EFV on lung cells, provoking the DDR pathway. Notably, the constitutive expression of this pathway (DDR) often leads to uncontrolled cell proliferation and eventually tumourigenesis, which could be the attribute of HAART components’ (such as EFV) effect on human cancers. Targeting the cell-cycle and its regulation holds a promising therapeutic intervention to the potential HAART associated carcinogenesis, particularly lung cancer.

Keywords: cell-cycle, DNA damage response, Efavirenz, lung cancer

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Authors: Juanita Wallace, Mark Jackson, Bethany Jurs

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Introduction: Exergames, VR games used as a form of exercise, are being used to reduce sedentary lifestyles in a vast number of populations. However, there is a distinct lack of research comparing the physiological response during VR exergaming to that of traditional exercises. The purpose of this study was to create a foundationary investigation establishing changes in physiological responses resulting from VR exergaming in a college aged population. Methods: In this IRB approved study, college aged students were recruited to play a virtual reality exergame (Beat Saber) on the Oculus Quest 2 (Facebook, 2021) in either a control group (CG) or training group (TG). Both groups consisted of subjects who were not habitual users of virtual reality. The CG played VR one time per week for three weeks and the TG played 150 min/week three weeks. Each group played the same nine Beat Saber songs, in a randomized order, during 30 minute sessions. Song difficulty was increased during play based on song performance. Subjects completed a pre- and posttests at which the following was collected: • Beat Saber Game Metrics: song level played, song score, number of beats completed per song and accuracy (beats completed/total beats) • Physiological Data: heart rate (max and avg.), active calories • Demographics Results: A total of 20 subjects completed the study; nine in the CG (3 males, 6 females) and 11 (5 males, 6 females) in the TG. • Beat Saber Song Metrics: The TG improved performance from a normal/hard difficulty to hard/expert. The CG stayed at the normal/hard difficulty. At the pretest there was no difference in game accuracy between groups. However, at the posttest the CG had a higher accuracy. • Physiological Data (Table 1): Average heart rates were similar between the TG and CG at both the pre- and posttest. However, the TG expended more total calories. Discussion: Due to the lack of peer reviewed literature on c exergaming using Beat Saber, the results of this study cannot be directly compared. However, the results of this study can be compared with the previously established trends for traditional exercise. In traditional exercise, an increase in training volume equates to increased efficiency at the activity. The TG should naturally increase in difficulty at a faster rate than the CG because they played 150 hours per week. Heart rate and caloric responses also increase during traditional exercise as load increases (i.e. speed or resistance). The TG reported an increase in total calories due to a higher difficulty of play. The song accuracy decreases in the TG can be explained by the increased difficulty of play. Conclusion: VR exergaming is comparable to traditional exercise for loads within the 50-70% of maximum heart rate. The ability to use VR for health could motivate individuals who do not engage in traditional exercise. In addition, individuals in health professions can and should promote VR exergaming as a viable way to increase physical activity and improve health in their clients/patients.

Keywords: virtual reality, exergaming, health, heart rate, wellness

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Authors: Jitka Rychtarikova

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Regional differences in mortality in the Czech Republic (CR) may be moderate from a broader European perspective, but important discrepancies in life expectancy can be found between smaller territorial units. In this study territorial units are based on Administrative Districts of Municipalities with Extended Powers (MEP). This definition came into force January 1, 2003. There are 205 units and the city of Prague. MEP represents the smallest unit for which mortality patterns based on life tables can be investigated and the Czech Statistical Office has been calculating such life tables (every five-years) since 2004. MEP life tables from 2009-2013 for males and females allowed the investigation of three main life cycles with the use of temporary life expectancies between the exact ages of 0 and 35; 35 and 65; and the life expectancy at exact age 65. The results showed regional survival inequalities primarily in adult and older ages. Consequently, only mortality indicators for adult and elderly population were related to census 2011 unlinked data for the same age groups. The most relevant socio-economic factors taken from the census are: having a partner, educational level and unemployment rate. The unemployment rate was measured for adults aged 35-64 completed years. Exploratory spatial data analysis methods were used to detect regional patterns in spatially contiguous units of MEP. The presence of spatial non-stationarity (spatial autocorrelation) of mortality levels for male and female adults (35-64), and elderly males and females (65+) was tested using global Moran’s I. Spatial autocorrelation of mortality patterns was mapped using local Moran’s I with the intention to depict clusters of low or high mortality and spatial outliers for two age groups (35-64 and 65+). The highest Moran’s I was observed for male temporary life expectancy between exact ages 35 and 65 (0.52) and the lowest was among women with life expectancy of 65 (0.26). Generally, men showed stronger spatial autocorrelation compared to women. The relationship between mortality indicators such as life expectancies and socio-economic factors like the percentage of males/females having a partner; percentage of males/females with at least higher secondary education; and percentage of unemployed males/females from economically active population aged 35-64 years, was evaluated using multiple regression (OLS). The results were then compared to outputs from geographically weighted regression (GWR). In the Czech Republic, there are two broader territories North-West Bohemia (NWB) and North Moravia (NM), in which excess mortality is well established. Results of the t-test of spatial regression showed that for males aged 30-64 the association between mortality and unemployment (when adjusted for education and partnership) was stronger in NM compared to NWB, while educational level impacted the length of survival more in NWB. Geographic variation and relationships in mortality of the CR MEP will also be tested using the spatial Durbin approach. The calculations were conducted by means of ArcGIS 10.6 and SAS 9.4.

Keywords: Czech Republic, mortality, municipality, socio-economic factors, spatial analysis

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Authors: D. Olszewska, J. Niewiedzial

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In some types of Li-ion batteries carbon in the form of graphite is used. Unfortunately, carbon materials, in particular graphite, have very good electrochemical properties, but increase their volume during charge/discharge cycles, which may even lead to an explosion of the cell. The cell element may be replaced by a composite material consisting of lithium-titanium oxide Li4Ti5O12 (LTO) modified with copper and nickel ions and carbon derived from sucrose. This way you can improve the conductivity of the material. LTO is appropriate only for applications which do not require high energy density because of its high operating voltage (ca. 1.5 V vs. Li/Li+). Specific capacity of Li4Ti5O12 is high enough for utilization in Li-ion batteries (theoretical capacity 175 mAh·g-1) but it is lower than capacity of graphite anodes. Materials based on Li4Ti5O12 do not change their volume during charging/discharging cycles, however, LTO has low conductivity. Another positive aspect of the use of sucrose in the carbon composite material is to eliminate the addition of carbon black from the anode of the battery. Therefore, the proposed materials contribute significantly to environmental protection and safety of selected lithium cells. New anode materials in order to obtain Li3.8Cu0.1Ni0.1Ti5O12 have been prepared by solid state synthesis using three-way: i) stoichiometric composition of Li2CO3, TiO2, CuO, NiO (A- Li3.8Cu0.1Ni0.1Ti5O12); ii) stoichiometric composition of Li2CO3, TiO2, Cu(NO3)2, Ni(NO3)2 (B-Li3.8Cu0.1Ni0.1Ti5O12); and iii) stoichiometric composition of Li2CO3, TiO2, CuO, NiO calcined with 10% of saccharose (Li3.8Cu0.1Ni0.1Ti5O12-C). Structure of materials was studied by X-ray diffraction (XRD). The electrochemical properties were performed using appropriately prepared cell Li|Li+|Li3.8Cu0.1Ni0.1Ti5O12 for cyclic voltammetry and discharge/charge measurements. The cells were periodically charged and discharged in the voltage range from 1.3 to 2.0 V applying constant charge/discharge current in order to determine the specific capacity of each electrode. Measurements at various values of the charge/discharge current (from C/10 to 5C) were carried out. Cyclic voltammetry investigation was carried out by applying to the cells a voltage linearly changing over time at a rate of 0.1 mV·s-1 (in the range from 2.0 to 1.3 V and from 1.3 to 2.0 V). The XRD method analyzes show that composite powders were obtained containing, in addition to the main phase, 4.78% and 4% TiO2 in A-Li3.8Cu0.1Ni0.1O12 and B-Li3.8Cu0.1Ni0.1O12, respectively. However, Li3.8Cu0.1Ni0.1O12-C material is three-phase: 63.84% of the main phase, 17.49 TiO2 and 18.67 Li2TiO3. Voltammograms of electrodes containing materials A-Li3.8Cu0.1Ni0.1O12 and B-Li3.8Cu0.1Ni0.1O12 are correct and repeatable. Peak cathode occurs for both samples at a potential approx. 1.52±0.01 V relative to a lithium electrode, while the anodic peak at potential approx. 1.65±0.05 V relative to a lithium electrode. Voltammogram of Li3.8Cu0.1Ni0.1Ti5O12-C (especially for the first measurement cycle) is not correct. There are large variations in values of specific current, which are not characteristic for materials LTO. From the point of view of safety and environmentally friendly production of Li-ion cells eliminating soot and applying Li3.8Cu0.1Ni0.1Ti5O12-C as an active material of an anode in lithium-ion batteries seems to be a good alternative to currently used materials.

Keywords: anode, Li-ion batteries, Li₄O₅O₁₂, spinel

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Authors: Ioanna Garefi, Irene Kalemaki

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Education is not solely about preparing young people for the world of work but also about equipping them with competences that will enable them to become socially proactive, empowered, responsible, and engaged citizens who will collectively contribute to and benefit from an inclusive and sustainable future. Hence, progress assessment towards competence development is an ongoing process where continuous efforts are needed. This paper abstract presents the work of the H2020 NEMESIS project that aims to investigate, experiment and co-create together with schools a model for introducing and embedding social innovation education (SIE henceforth) in European primary and secondary schools. All in all, during the 2018-2019 academic year, 8 schools from 5 European countries involving 56 teachers, 1030 students, and 80 external stakeholders, experimented with different methodologies for embedding SIE in their contexts. This paper captures briefly the impact of these efforts towards the cultivation and progression of students’ social innovation (SI henceforth) competences. As part of the model, 14 SI competences, whose progress was evaluated, have been introduced falling under 3 interrelated categories: competences for identifying opportunities for social and collective value creation, competences for developing collaborations and building meaningful relations and competences for taking action both on an individual and collective level. Methodologically wise, the evaluation strategy employed was informed by a realist approach, enabling the researchers to go beyond synthesizing 'what happened' and towards understanding 'why it happened', delving into ‘what works, for whom and in what circumstances’. The reason for choosing such an approach was because it goes beyond attempting to answer the basic yes or no question of evaluation and focus on an ‘explanatory quest’ tracing the limits of when and where intervention is effective. A rich mix of sources of evidence have been employed, from focus groups with 80 people from the 5 EU countries to an online survey to 206 students, classroom observations, students’ narratives granting them with the opportunity to freely express their opinions, short stories letting students express their feelings through their imagination and also, drawings so that younger children can express their perception of reality. All these evidences offered insights on the impact of SIE on the development of students’ competences. Research findings showed that students progressed in all 14 SI competences through their involvement in the different activities. This positive progression is attributed to the model’s three core principles: 1) the student-centered approach, rendering students active and self-determined producers of their own learning, 2) the co-creation process fostering intergenerational interactions, empowering thus students by making their voices heard and valued and also, 3) the transformative social action whereby through their projects, students are able to witness the impact they are bringing about with their actions. Concluding, these initial findings, together with the forthcoming evaluation research to a pool of 30 schools around Europe, have the potential to raise the dynamics of the under-investigated field of SIE and encourage its embeddedness in more schools around Europe.

Keywords: competence development, education, social innovation, students

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Authors: Maria Fatima Almeida, Rodrigo Calili, George Soares, João Krause, Myrthes Marcele Dos Santos, Anna Carolina Suzano E. Silva, Marcos Alexandre Da

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In Brazil, the building sector accounts for 1/6 of energy consumption and 50% of electricity consumption. A complex sector with several driving actors plays an essential role in the country's economy. Currently, the digitalization readiness in this sector is still low, mainly due to the high investment costs and the difficulty of estimating the benefits of digital technologies in buildings. Nevertheless, the potential contribution of digitalization for increasing energy efficiency in the building sector in Brazil has been pointed out as relevant in the political and sectoral contexts, both in the medium and long-term horizons. To contribute to the debate on the possible evolving trajectories of digitalization in the building sector in Brazil and to subsidize the formulation or revision of current public policies and managerial decisions, three future scenarios were created to anticipate the potential energy efficiency in the building sector in Brazil due to digitalization by 2050. This work aims to present these scenarios as a basis to foresight the potential energy efficiency in this sector, according to different digitalization paces - slow, moderate, or fast in the 2050 horizon. A methodological approach was proposed to create alternative prospective scenarios, combining the Global Business Network (GBN) and the Laboratory for Investigation in Prospective Strategy and Organisation (LIPSOR) methods. This approach consists of seven steps: (i) definition of the question to be foresighted and time horizon to be considered (2050); (ii) definition and classification of a set of key variables, using the prospective structural analysis; (iii) identification of the main actors with an active role in the digital and energy spheres; (iv) characterization of the current situation (2021) and identification of main uncertainties that were considered critical in the development of alternative future scenarios; (v) scanning possible futures using morphological analysis; (vi) selection and description of the most likely scenarios; (vii) foresighting the potential energy efficiency in each of the three scenarios, namely slow digitalization; moderate digitalization, and fast digitalization. Each scenario begins with a core logic and then encompasses potentially related elements, including potential energy efficiency. Then, the first scenario refers to digitalization at a slow pace, with induction by the government limited to public buildings. In the second scenario, digitalization is implemented at a moderate pace, induced by the government in public, commercial, and service buildings, through regulation integrating digitalization and energy efficiency mechanisms. Finally, in the third scenario, digitalization in the building sector is implemented at a fast pace in the country and is strongly induced by the government, but with broad participation of private investments and accelerated adoption of digital technologies. As a result of the slow pace of digitalization in the sector, the potential for energy efficiency stands at levels below 10% of the total of 161TWh by 2050. In the moderate digitalization scenario, the potential reaches 20 to 30% of the total 161TWh by 2050. Furthermore, in the rapid digitalization scenario, it will reach 30 to 40% of the total 161TWh by 2050.

Keywords: building digitalization, energy efficiency, scenario building, prospective structural analysis, morphological analysis

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Authors: J. Gabrielczyk, J. Kluitmann, T. Dammeyer, H. J. Jördening

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High-level expression of proteins in bacteria often causes production of insoluble protein aggregates, called inclusion bodies (IB). They contain mainly one type of protein and offer an easy and efficient way to get purified protein. On the other hand, proteins in IB are normally devoid of function and therefore need a special treatment to become active. Most refolding techniques aim at diluting the solubilizing chaotropic agents. Unfortunately, optimal refolding conditions have to be found empirically for every protein. For large-scale applications, a simple refolding process with high yields and high final enzyme concentrations is still missing. The constructed plasmid pASK-IBA63b containing the sequence of fructosyltransferase (FTF, EC 2.4.1.162) from Bacillus subtilis NCIMB 11871 was transformed into E. coli BL21 (DE3) Rosetta. The bacterium was cultivated in a fed-batch bioreactor. The produced FTF was obtained mainly as IB. For refolding experiments, five different amounts of IBs were solubilized in urea buffer with protein concentration of 0.2-8.5 g/L. Solubilizates were refolded with batch or continuous dialysis. The refolding yield was determined by measuring the protein concentration of the clear supernatant before and after the dialysis. Particle size was measured by dynamic light scattering. We tested the solubilization properties of fructosyltransferase IBs. The particle size measurements revealed that the solubilization of the aggregates is achieved at urea concentration of 5M or higher and confirmed by absorption spectroscopy. All results confirm previous investigations that refolding yields are dependent upon initial protein concentration. In batch dialysis, the yields dropped from 67% to 12% and 72% to 19% for continuous dialysis, in relation to initial concentrations from 0.2 to 8.5 g/L. Often used additives such as sucrose and glycerol had no effect on refolding yields. Buffer screening indicated a significant increase in activity but also temperature stability of FTF with citrate/phosphate buffer. By adding citrate to the dialysis buffer, we were able to increase the refolding yields to 82-47% in batch and 90-74% in the continuous process. Further experiments showed that in general, higher ionic strength of buffers had major impact on refolding yields; doubling the buffer concentration increased the yields up to threefold. Finally, we achieved corresponding high refolding yields by reducing the chamber volume by 75% and the amount of buffer needed. The refolded enzyme had an optimal activity of 12.5±0.3 x104 units/g. However, detailed experiments with native FTF revealed a reaggregation of the molecules and loss in specific activity depending on the enzyme concentration and particle size. For that reason, we actually focus on developing a process of simultaneous enzyme refolding and immobilization. The results of this study show a new approach in finding optimal refolding conditions for inclusion bodies at high concentrations. Straightforward buffer screening and increase of the ionic strength can optimize the refolding yield of the target protein by 400%. Gentle removal of chaotrope with continuous dialysis increases the yields by an additional 65%, independent of the refolding buffer applied. In general time is the crucial parameter for successful refolding of solubilized proteins.

Keywords: dialysis, inclusion body, refolding, solubilization

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