Search results for: social autopsy research design
Commenced in January 2007
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Paper Count: 37197

Search results for: social autopsy research design

567 Temperature Dependence of the Optoelectronic Properties of InAs(Sb)-Based LED Heterostructures

Authors: Antonina Semakova, Karim Mynbaev, Nikolai Bazhenov, Anton Chernyaev, Sergei Kizhaev, Nikolai Stoyanov

Abstract:

At present, heterostructures are used for fabrication of almost all types of optoelectronic devices. Our research focuses on the optoelectronic properties of InAs(Sb) solid solutions that are widely used in fabrication of light emitting diodes (LEDs) operating in middle wavelength infrared range (MWIR). This spectral range (2-6 μm) is relevant for laser diode spectroscopy of gases and molecules, for systems for the detection of explosive substances, medical applications, and for environmental monitoring. The fabrication of MWIR LEDs that operate efficiently at room temperature is mainly hindered by the predominance of non-radiative Auger recombination of charge carriers over the process of radiative recombination, which makes practical application of LEDs difficult. However, non-radiative recombination can be partly suppressed in quantum-well structures. In this regard, studies of such structures are quite topical. In this work, electroluminescence (EL) of LED heterostructures based on InAs(Sb) epitaxial films with the molar fraction of InSb ranging from 0 to 0.09 and multi quantum-well (MQW) structures was studied in the temperature range 4.2-300 K. The growth of the heterostructures was performed by metal-organic chemical vapour deposition on InAs substrates. On top of the active layer, a wide-bandgap InAsSb(Ga,P) barrier was formed. At low temperatures (4.2-100 K) stimulated emission was observed. As the temperature increased, the emission became spontaneous. The transition from stimulated emission to spontaneous one occurred at different temperatures for structures with different InSb contents in the active region. The temperature-dependent carrier lifetime, limited by radiative recombination and the most probable Auger processes (for the materials under consideration, CHHS and CHCC), were calculated within the framework of the Kane model. The effect of various recombination processes on the carrier lifetime was studied, and the dominant role of Auger processes was established. For MQW structures quantization energies for electrons, light and heavy holes were calculated. A characteristic feature of the experimental EL spectra of these structures was the presence of peaks with energy different from that of calculated optical transitions between the first quantization levels for electrons and heavy holes. The obtained results showed strong effect of the specific electronic structure of InAsSb on the energy and intensity of optical transitions in nanostructures based on this material. For the structure with MQWs in the active layer, a very weak temperature dependence of EL peak was observed at high temperatures (>150 K), which makes it attractive for fabricating temperature-resistant gas sensors operating in the middle-infrared range.

Keywords: Electroluminescence, InAsSb, light emitting diode, quantum wells

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566 Cytotoxicity and Genotoxicity of Glyphosate and Its Two Impurities in Human Peripheral Blood Mononuclear Cells

Authors: Marta Kwiatkowska, Paweł Jarosiewicz, Bożena Bukowska

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Glyphosate (N-phosphonomethylglycine) is a non-selected broad spectrum ingredient in the herbicide (Roundup) used for over 35 years for the protection of agricultural and horticultural crops. Glyphosate was believed to be environmentally friendly but recently, a large body of evidence has revealed that glyphosate can negatively affect on environment and humans. It has been found that glyphosate is present in the soil and groundwater. It can also enter human body which results in its occurrence in blood in low concentrations of 73.6 ± 28.2 ng/ml. Research conducted for potential genotoxicity and cytotoxicity can be an important element in determining the toxic effect of glyphosate. Due to regulation of European Parliament 1107/2009 it is important to assess genotoxicity and cytotoxicity not only for the parent substance but also its impurities, which are formed at different stages of production of major substance – glyphosate. Moreover verifying, which of these compounds are more toxic is required. Understanding of the molecular pathways of action is extremely important in the context of the environmental risk assessment. In 2002, the European Union has decided that glyphosate is not genotoxic. Unfortunately, recently performed studies around the world achieved results which contest decision taken by the committee of the European Union. World Health Organization (WHO) in March 2015 has decided to change the classification of glyphosate to category 2A, which means that the compound is considered to "probably carcinogenic to humans". This category relates to compounds for which there is limited evidence of carcinogenicity to humans and sufficient evidence of carcinogenicity on experimental animals. That is why we have investigated genotoxicity and cytotoxicity effects of the most commonly used pesticide: glyphosate and its impurities: N-(phosphonomethyl)iminodiacetic acid (PMIDA) and bis-(phosphonomethyl)amine on human peripheral blood mononuclear cells (PBMCs), mostly lymphocytes. DNA damage (analysis of DNA strand-breaks) using the single cell gel electrophoresis (comet assay) and ATP level were assessed. Cells were incubated with glyphosate and its impurities: PMIDA and bis-(phosphonomethyl)amine at concentrations from 0.01 to 10 mM for 24 hours. Evaluating genotoxicity using the comet assay showed a concentration-dependent increase in DNA damage for all compounds studied. ATP level was decreased to zero as a result of using the highest concentration of two investigated impurities, like bis-(phosphonomethyl)amine and PMIDA. Changes were observed using the highest concentration at which a person can be exposed as a result of acute intoxication. Our survey leads to a conclusion that the investigated compounds exhibited genotoxic and cytotoxic potential but only in high concentrations, to which people are not exposed environmentally. Acknowledgments: This work was supported by the Polish National Science Centre (Contract-2013/11/N/NZ7/00371), MSc Marta Kwiatkowska, project manager.

Keywords: cell viability, DNA damage, glyphosate, impurities, peripheral blood mononuclear cells

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565 Functional Switching of Serratia marcescens Transcriptional Regulator from Activator to Inhibitor of Quorum Sensing by Exogenous Addition

Authors: Norihiro Kato, Yuriko Takayama

Abstract:

Some gram-negative bacteria enable the simultaneous activation of gene expression involved in N-acylhomoserine lactone (AHL) dependent cell-to-cell communication system. Such regulatory system for the bacterial group behavior is termed as quorum sensing (QS) because a diffusible AHL signal can accumulate around the cell during the increase of the cell density and trigger activation of the sequential QS process. By blocking the QS, the expression of diverse genes related to infection, antibiotic production, and biofilm formation is inhibited. Conditioning of QS by regulation of the DNA-receptor-AHL interaction is a potential target for enhancing host defenses against pathogenicity. We focused on engineered application of transcriptional regulator SpnR produced in opportunistic human pathogen Serratia marcescens. The SpnR can interact with AHL signals at an N-terminal domain and also with a promoter region of a QS target gene at a C-terminal domain. As the initial process of the QS activation, the SpnR forms a complex with the AHL to enhance the expression of pig cluster; the SpnR normally acts as an activator for the expression of the QS-dependent gene. In this research, we attempt to artificially control QS by changing the role of SpnR. The QS-dependent prodigiosin production is expected to inhibit by externally added SpnR in the culture broth of AS-1 strain because the AHL concentration was kept below the threshold by AHL-SpnR complex formation. Maltose-binding protein (MBP)-tagged SpnR (MBP-SpnR) was overexpressed in Escherichia coli and purified using an affinity chromatography equipped with an amylose resin column. The specific interaction between AHL and MBP-SpnR was demonstrated by quartz crystal microbalance (QCM) sensor. AHL with amino end-group was coupled with COOH-terminated self-assembled monolayer prepared on a gold electrode of 27-MHz quartz crystal sensor using water-soluble carbodiimide. After the injection of MBP-SpnR into a cup-type sensor cell filled with the buffer solution, time course of resonant frequency change (ΔFs) was determined. A decrease of ΔFs clearly showed the uptake of MBP-SpnR onto the AHL-immobilized electrode. Furthermore, no binding affinity was observed after the heat-inactivation of MBP-SpnR at 80ºC. These results suggest that MBP-SpnR possesses a specific affinity for AHL. MBP-SpnR was added to the culture medium as an AHL trap to study inhibitory effects on intracellularly accumulated prodigiosin. With approximately 2 µM MBP-SpnR, the amount of prodigiosin induced was half that of the control without any additives. In conclusion, the function of SpnR could be switched by adding it to the cell culture. Exogenously added MBP-SpnR possesses high affinity for AHL derived from cells and acts as an inhibitor of AHL-mediated QS.

Keywords: intracellular signaling, microbial biotechnology, quorum sensing, transcriptional regulator

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564 The Charge Exchange and Mixture Formation Model in the ASz-62IR Radial Aircraft Engine

Authors: Pawel Magryta, Tytus Tulwin, Paweł Karpiński

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The ASz62IR engine is a radial aircraft engine with 9 cylinders. This object is produced by the Polish company WSK "PZL-KALISZ" S.A. This is engine is currently being developed by the above company and Lublin University of Technology. In order to provide an effective work of the technological development of this unit it was decided to made the simulation model. The model of ASz-62IR was developed with AVL BOOST software which is a tool dedicated to the one-dimensional modeling of internal combustion engines. This model can be used to calculate parameters of an air and fuel flow in an intake system including charging devices as well as combustion and exhaust flow to the environment. The main purpose of this model is the analysis of the charge exchange and mixture formation in this engine. For this purpose, the model consists of elements such: as air inlet, throttle system, compressor connector, charging compressor, inlet pipes and injectors, outlet pipes, fuel injection and model of fuel mixing and evaporation. The model of charge exchange and mixture formation was based on the model of mass flow rate in intake and exhaust pipes, and also on the calculation of gas properties values like gas constant or thermal capacity. This model was based on the equations to describe isentropic flow. The energy equation to describe flow under steady conditions was transformed into the mass flow equation. In the model the flow coefficient μσ was used, that varies with the stroke/valve opening and was determined in a steady flow state. The geometry of the inlet channels and other key components was mapped with reference to the technical documentation of the engine and empirical measurements of the structure elements. The volume of elements on the charge flow path between the air inlet and the exhaust outlet was measured by the CAD mapping of the structure. Taken from the technical documentation, the original characteristics of the compressor engine was entered into the model. Additionally, the model uses a general model for the transport of chemical compounds of the mixture. There are 7 compounds used, i.e. fuel, O2, N2, CO2, H2O, CO, H2. A gasoline fuel of a calorific value of 43.5 MJ/kg and an air mass fraction for stoichiometric mixture of 14.5 were used. Indirect injection into the intake manifold is used in this model. The model assumes the following simplifications: the mixture is homogenous at the beginning of combustion, accordingly, mixture stoichiometric coefficient A/F remains constant during combustion, combusted and non-combusted charges show identical pressures and temperatures although their compositions change. As a result of the simulation studies based on the model described above, the basic parameters of combustion process, charge exchange, mixture formation in cylinders were obtained. The AVL Boost software is very useful for the piston engine performance simulations. This work has been financed by the Polish National Centre for Research and Development, INNOLOT, under Grant Agreement No. INNOLOT/I/1/NCBR/2013.

Keywords: aviation propulsion, AVL Boost, engine model, charge exchange, mixture formation

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563 Assessment of the Impact of Regular Pilates Exercises on Static Balance in Healthy Adult Women: Preliminary Report

Authors: Anna Słupik, Krzysztof Jaworski, Anna Mosiołek, Dariusz Białoszewski

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Background: Maintaining the correct body balance is essential in the prevention of falls in the elderly, which is especially important for women because of postmenopausal osteoporosis and the serious consequences of falls. One of the exercise methods which is very popular among adults, and which may affect body balance in a positive way is the pilates method. The aim of the study was to evaluate the effect of regular pilates exercises on the ability to maintain body balance in static conditions in adult healthy women. Material and methods: The study group consisted of 20 healthy women attending pilates twice a week for at least 1 year. The control group consisted of 20 healthy women physically inactive. Women in the age range from 35 to 50 years old without pain in musculoskeletal system or other pain were only qualified to the groups. Body balance was assessed using MatScan VersaTek platform with Sway Analysis Module based on Matscan Clinical 6.7 software. The balance was evaluated under the following conditions: standing on both feet with eyes open, standing on both feet with eyes closed, one-leg standing (separately on the right and left foot) with eyes open. Each test lasted 30 seconds. The following parameters were calculated: estimated size of the ellipse of 95% confidence, the distance covered by the Center of Gravity (COG), the size of the maximum shift in the sagittal and frontal planes and load distribution between the left and right foot, as well as between rear- and forefoot. Results: It was found that there is significant difference between the groups in favor of the study group in the size of the confidence ellipse and maximum shifts of COG in the sagittal plane during standing on both feet, both with the eyes open and closed (p < 0.05). While standing on one leg both on the right and left leg, with eyes opened there was a significant difference in favor of the study group, in terms of the size of confidence ellipse, the size of the maximum shifts in the sagittal and in the frontal plane (p < 0.05). There were no differences between the distribution of load between the right and left foot (standing with both feet), nor between fore- and rear foot (in standing with both feet or one-leg). Conclusions: 1. Static balance in women exercising regularly by pilates method is better than in inactive women, which may in the future prevent falls and their consequences. 2. The observed differences in maintaining balance in frontal plane in one-leg standing may indicate a positive impact of pilates exercises on the ability to maintain global balance in terms of the reduced support surface. 3. Pilates method can be used as a form preventive therapy for all people who are expected to have problems with body balance in the future, for example in chronic neurological disorders or vestibular problems. 4. The results have shown that further prospective randomized research on a larger and more representative group is needed.

Keywords: balance exercises, body balance, pilates, pressure distribution, women

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562 The Relationship between Wasting and Stunting in Young Children: A Systematic Review

Authors: Susan Thurstans, Natalie Sessions, Carmel Dolan, Kate Sadler, Bernardette Cichon, Shelia Isanaka, Dominique Roberfroid, Heather Stobagh, Patrick Webb, Tanya Khara

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For many years, wasting and stunting have been viewed as separate conditions without clear evidence supporting this distinction. In 2014, the Emergency Nutrition Network (ENN) examined the relationship between wasting and stunting and published a report highlighting the evidence for linkages between the two forms of undernutrition. This systematic review aimed to update the evidence generated since this 2014 report to better understand the implications for improving child nutrition, health and survival. Following PRISMA guidelines, this review was conducted using search terms to describe the relationship between wasting and stunting. Studies related to children under five from low- and middle-income countries that assessed both ponderal growth/wasting and linear growth/stunting, as well as the association between the two, were included. Risk of bias was assessed in all included studies using SIGN checklists. 45 studies met the inclusion criteria- 39 peer reviewed studies, 1 manual chapter, 3 pre-print publications and 2 published reports. The review found that there is a strong association between the two conditions whereby episodes of wasting contribute to stunting and, to a lesser extent, stunting leads to wasting. Possible interconnected physiological processes and common risk factors drive an accumulation of vulnerabilities. Peak incidence of both wasting and stunting was found to be between birth and three months. A significant proportion of children experience concurrent wasting and stunting- Country level data suggests that up to 8% of children under 5 may be both wasted and stunted at the same time, global estimates translate to around 16 million children. Children with concurrent wasting and stunting have an elevated risk of mortality when compared to children with one deficit alone. These children should therefore be considered a high-risk group in the targeting of treatment. Wasting, stunting and concurrent wasting and stunting appear to be more prevalent in boys than girls and it appears that concurrent wasting and stunting peaks between 12- 30 months of age with younger children being the most affected. Seasonal patterns in prevalence of both wasting and stunting are seen in longitudinal and cross sectional data and in particular season of birth has been shown to have an impact on a child’s subsequent experience of wasting and stunting. Evidence suggests that the use of mid-upper-arm circumference combined with weight-for-age Z-score might effectively identify children most at risk of near-term mortality, including those concurrently wasted and stunted. Wasting and stunting frequently occur in the same child, either simultaneously or at different moments through their life course. Evidence suggests there is a process of accumulation of nutritional deficits and therefore risk over the life course of a child demonstrates the need for a more integrated approach to prevention and treatment strategies to interrupt this process. To achieve this, undernutrition policies, programmes, financing and research must become more unified.

Keywords: Concurrent wasting and stunting, Review, Risk factors, Undernutrition

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561 Ukrainians Professors in a Luso-Hispanophone Brazilian Border Region: a Case-Study on the Management of Multilingualism in Higher Education

Authors: Isis Ribeiro Berger

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In view of recent war conflicts between Russia and Ukraine, the government of Paraná State, in Brazil, started a program to host Ukrainian scientists in state universities in 2022. The initiative aimed at integrating these scientists into the Brazilian academic community, strengthening the role of universities in producing science and innovation even in times of war, as well as fostering Higher Education internationalization. Paraná state was a pioneer in this initiative due to the fact it has been home to the largest contingent of immigrants and descendants of Ukrainians in Brazil because of migratory processes that began at the end of the 19th century. One of the universities receiving Ukrainian scientists is in Foz do Iguaçu, a city that borders Argentina and Paraguay. It is a multilingual environment, whose majority languages are Portuguese (the official language of Brazil), Spanish (the official language of both Argentina and Paraguay), as well as Guarani (the co-official indigenous language of Paraguay). It is in such a sociolinguistic environment that two Ukrainian professors began their activities within the scope of an Interdisciplinary Postgraduate Program (master’s and doctorate degree). This case study, whose theme is the management of multilingualism, was developed within the scope of Language Policy. It aimed at identifying the attitudes of both Ukrainian professors and postgraduate students towards multilingualism in this context, given the plural linguistic repertoire of the academic community, as well as identifying the language management strategies for the construction of knowledge implemented by the program and in the classroom by these participants. Therefore, the study was conducted under a qualitative approach, for which surveys and interviews were adopted as part of its methodological procedures. Data revealed the presence of different languages in the classroom (Portuguese, Spanish, English and Ukrainian), which made pedagogical practices challenging for both professors and students, whose levels of knowledge in the different languages varied significantly. The results indicate that multilingualism was the norm as the means of instruction adopted in this context, in which bilingual Portuguese-English-Ukrainian instruction was used by the professors in their lectures. Although English has been privileged for the internationalization of Higher Education in various contexts, it was not used as an exclusive means of instruction in this case, mostly because it is a predominantly Portuguese-Spanish-speaking environment. In addition, the professors counted on the mediation of an interpreter hired by the program since not every student had sufficient knowledge of English as part of their repertoires. The findings also suggest Portuguese is the language that most of the participants of this study prefer, both because it is the mother tongue of majority, and because it is the official language of the host country to the professors, who have sought to integrate to the local culture and community. This research is inserted in the Axis: Multilingualism and Education, of the UNESCO Chair on Language Policies for Multilingualism to which this study is related.

Keywords: attitudes, border region, multilingualism management, Ukrainian professors

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560 Coordinative Remote Sensing Observation Technology for a High Altitude Barrier Lake

Authors: Zhang Xin

Abstract:

Barrier lakes are lakes formed by storing water in valleys, river valleys or riverbeds after being blocked by landslide, earthquake, debris flow, and other factors. They have great potential safety hazards. When the water is stored to a certain extent, it may burst in case of strong earthquake or rainstorm, and the lake water overflows, resulting in large-scale flood disasters. In order to ensure the safety of people's lives and property in the downstream, it is very necessary to monitor the barrier lake. However, it is very difficult and time-consuming to manually monitor the barrier lake in high altitude areas due to the harsh climate and steep terrain. With the development of earth observation technology, remote sensing monitoring has become one of the main ways to obtain observation data. Compared with a single satellite, multi-satellite remote sensing cooperative observation has more advantages; its spatial coverage is extensive, observation time is continuous, imaging types and bands are abundant, it can monitor and respond quickly to emergencies, and complete complex monitoring tasks. Monitoring with multi-temporal and multi-platform remote sensing satellites can obtain a variety of observation data in time, acquire key information such as water level and water storage capacity of the barrier lake, scientifically judge the situation of the barrier lake and reasonably predict its future development trend. In this study, The Sarez Lake, which formed on February 18, 1911, in the central part of the Pamir as a result of blockage of the Murgab River valley by a landslide triggered by a strong earthquake with magnitude of 7.4 and intensity of 9, is selected as the research area. Since the formation of Lake Sarez, it has aroused widespread international concern about its safety. At present, the use of mechanical methods in the international analysis of the safety of Lake Sarez is more common, and remote sensing methods are seldom used. This study combines remote sensing data with field observation data, and uses the 'space-air-ground' joint observation technology to study the changes in water level and water storage capacity of Lake Sarez in recent decades, and evaluate its safety. The situation of the collapse is simulated, and the future development trend of Lake Sarez is predicted. The results show that: 1) in recent decades, the water level of Lake Sarez has not changed much and remained at a stable level; 2) unless there is a strong earthquake or heavy rain, it is less likely that the Lake Sarez will be broken under normal conditions, 3) lake Sarez will remain stable in the future, but it is necessary to establish an early warning system in the Lake Sarez area for remote sensing of the area, 4) the coordinative remote sensing observation technology is feasible for the high altitude barrier lake of Sarez.

Keywords: coordinative observation, disaster, remote sensing, geographic information system, GIS

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559 The Burmese Exodus of 1942: Towards Evolving Policy Protocols for a Refugee Archive

Authors: Vinod Balakrishnan, Chrisalice Ela Joseph

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The Burmese Exodus of 1942, which left more than 4 lakh as refugees and thousands dead, is one of the worst forced migrations in recorded history. Adding to the woes of the refugees is the lack of credible documentation of their lived experiences, trauma, and stories and their erasure from recorded history. Media reports, national records, and mainstream narratives that have registered the exodus provide sanitized versions which have reduced the refugees to a nameless, faceless mass of travelers and obliterated their lived experiences, trauma, and sufferings. This attitudinal problem compels the need to stem the insensitivity that accompanies institutional memory by making a case for a more humanistically evolved policy that puts in place protocols for the way the humanities would voice the concern for the refugee. A definite step in this direction and a far more relevant project in our times is the need to build a comprehensive refugee archive that can be a repository of the refugee experiences and perspectives. The paper draws on Hannah Arendt’s position on the Jewish refugee crisis, Agamben’s work on statelessness and citizenship, Foucault’s notion of governmentality and biopolitics, Edward Said’s concepts on Exile, Fanon’s work on the dispossessed, Derrida’s work on ‘the foreigner and hospitality’ in order to conceptualize the refugee condition which will form the theoretical framework for the paper. It also refers to the existing scholarship in the field of refugee studies such as Roger Zetter’s work on the ‘refugee label’, Philip Marfleet’s work on ‘refugees and history’, Lisa Malkki’s research on the anthropological discourse of the refugee and refugee studies. The paper is also informed by the work that has been done by the international organizations to address the refugee crisis. The emphasis is on building a strong argument for the establishment of the refugee archive that finds but a passing and a none too convincing reference in refugee studies in order to enable a multi-dimensional understanding of the refugee crisis. Some of the old questions cannot be dismissed as outdated as the continuing travails of the refugees in different parts of the world only remind us that they are still, largely, unanswered. The questions are -What is the nature of a Refugee Archive? How is it different from the existing historical and political archives? What are the implications of the refugee archive? What is its contribution to refugee studies? The paper draws on Diana Taylor’s concept of the archive and the repertoire to theorize the refugee archive as a repository that has the documentary function of the ‘archive’ and the ‘agency’ function of the repertoire. It then reads Ayya’s Accounts- a memoir by Anand Pandian -in the light of Hannah Arendt’s concepts of the ‘refugee as vanguard’ and ‘story telling as political action’- to illustrate how the memoir contributes to the refugee archive that provides the refugee a place and agency in history. The paper argues for a refugee archive that has implications for the formulation of inclusive refugee policies.

Keywords: Ayya’s Accounts, Burmese Exodus, policy protocol, refugee archive

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558 Ascidian Styela rustica Proteins’ Structural Domains Predicted to Participate in the Tunic Formation

Authors: M. I. Tyletc, O. I. Podgornya, T. G. Shaposhnikova, S. V. Shabelnikov, A. G. Mittenberg, M. A. Daugavet

Abstract:

Ascidiacea is the most numerous class of the Tunicata subtype. These chordates' distinctive feature of the anatomical structure is a tunic consisting of cellulose fibrils, protein molecules, and single cells. The mechanisms of the tunic formation are not known in detail; tunic formation could be used as the model system for studying the interaction of cells with the extracellular matrix. Our model species is the ascidian Styela rustica, which is prevalent in benthic communities of the White Sea. As previously shown, the tunic formation involves morula blood cells, which contain the major 48 kDa protein p48. P48 participation in the tunic formation was proved using antibodies against the protein. The nature of the protein and its function remains unknown. The current research aims to determine the amino acid sequence of p48, as well as to clarify its role in the tunic formation. The peptides that make up the p48 amino acid sequence were determined by mass spectrometry. A search for peptides in protein sequence databases identified sequences homologous to p48 in Styela clava, Styela plicata, and Styela canopus. Based on sequence alignment, their level of similarity was determined as 81-87%. The correspondent sequence of ascidian Styela canopus was used for further analysis. The Styela rustica p48 sequence begins with a signal peptide, which could indicate that the protein is secretory. This is consistent with experimentally obtained data: the contents of morula cells secreted in the tunic matrix. The isoelectric point of p48 is 9.77, which is consistent with the experimental results of acid electrophoresis of morula cell proteins. However, the molecular weight of the amino acid sequence of ascidian Styela canopus is 103 kDa, so p48 of Styela rustica is a shorter homolog. The search for conservative functional domains revealed the presence of two Ca-binding EGF-like domains, thrombospondin (TSP1) and tyrosinase domains. The p48 peptides determined by mass spectrometry fall into the region of the sequence corresponding to the last two domains and have amino acid substitutions as compared to Styela canopus homolog. The tyrosinase domain (pfam00264) is known to be part of the phenoloxidase enzyme, which participates in melanization processes and the immune response. The thrombospondin domain (smart00209) interacts with a wide range of proteins, and is involved in several biological processes, including coagulation, cell adhesion, modulation of intercellular and cell-matrix interactions, angiogenesis, wound healing and tissue remodeling. It can be assumed that the tyrosinase domain in p48 plays the role of the phenoloxidase enzyme, and TSP1 provides a link between the extracellular matrix and cell surface receptors, and may also be responsible for the repair of the tunic. The results obtained are consistent with experimental data on p48. The domain organization of protein suggests that p48 is an enzyme involved in the tunic tunning and is an important regulator of the organization of the extracellular matrix.

Keywords: ascidian, p48, thrombospondin, tyrosinase, tunic, tunning

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557 Migratory Diaspora: The Media and the Human Element

Authors: Peter R. Alfieri

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The principal aim of this research and presentation is to give global and personal perspective of the migratory diaspora and how it is perceived by a substantial majority that relies on the media’s portrayal of migratory movements. Since its Greek origins the word “diaspora” has taken on several connotations, but none has surpassed its use in regard to the human element; because since before the dawn of history, man has had to struggle for survival. That survival was a struggle against the elements and other natural enemies, but none as tenacious and relentless as other men. Many have used the term diaspora to describe the spread of certain ethnic groups resulting in new generations in new places; but has the human diaspora been as haphazard as that of spores? The quest for survival has spawned migrations that are not quite that simple, even though it has several similarities to plant spores or dandelion seeds flying throughout the atmosphere. Man kind has constantly migrated in search of food, shelter, and safety. When they were able to find food and shelter, they would inform others who would venture to the new place. Information, whether through word of mouth, written material, or visual communications, has been a moving force in man’s life; and it spurred migrants in their quest for better environments. Today we pride ourselves in being able to communicate instantly with anyone anywhere in the world, and we are privileged to see most of what is happening in the world thanks to the highly developed modern media. Is Media a “wind/force” instrumental in propelling the diaspora throughout the world? The media has been the tool that has incentive many migratory, but unfortunately it is also the means responsible for many misconceptions regarding migrants and their hosts. Has the Media presented an unbiased view of the migrant or has it been the means that generated negative or prejudiced views of the migrant and, perhaps, the host environment? Some examples were easily seen in 19th century the United States where they advertised the following, “Help needed, Irish need not apply”. How do immigrants circumvent latent barriers that are not as obvious as the ones just mentioned? Some immigrants return home and have children that decide to emigrate. It is a perpetual cycle in the search for self-improvement. The stories that are brought back might be inspiration for the new generation of emigrants. Poverty, hunger, and political turmoil spur most migrations. The majority learn from others or through the media about certain destinations that will provide one or several opportunities to improve their existence. Many of those migrants suffer untold hardships to succeed. When they succeed, they provide a great incentive for their children to obtain an education or skill that will insure them a better life. Although the new environment may contribute greatly to a successful career, most immigrants do not forget their own struggle. They see the media’s portrayal of other migrants from all over the globe. Some try to communicate to others the true feelings of despair felt by immigrants, because they are all brothers and sisters in the perennial struggle for a better life. “HOPE” for a better life drives the immigrant toward the unknown and it has helped overcome the obstacles that present themselves challenging every newcomer. Hope and perseverance strengthen the resolve of the migrant in his struggle to survive.

Keywords: media, migration, heath, education, obstacles

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556 The Improved Therapeutic Effect of Trans-Cinnamaldehyde on Adipose-Derived Stem Cells without Chemical Induction

Authors: Karthyayani Rajamani, Yi-Chun Lin, Tung-Chou Wen, Jeanne Hsieh, Yi-Maun Subeq, Jen-Wei Liu, Po-Cheng Lin, Horng-Jyh Harn, Shinn-Zong Lin, Tzyy-Wen Chiou

Abstract:

Assuring cell quality is an essential parameter for the success of stem cell therapy, utilization of various components to improve this potential has been the primary goal of stem cell research. The aim of this study was not only to demonstrate the capacity of trans-cinnamaldehyde (TC) to reverse stress-induced senescence but also improve the therapeutic abilities of stem cells. Because of the availability and the promising application potential in regenerative medicine, adipose-derived stem cells (ADSCs) were chosen for the study. We found that H2O2 treatment resulted in the expression of senescence characteristics in the ADSCs, including decreased proliferation rate, increased senescence-associated- β-galactosidase (SA-β-gal) activity, decreased SIRT1 (silent mating type information regulation 2 homologs) expression and decreased telomerase activity. However, TC treatment was sufficient to rescue or reduce the effects of H2O2 induction, ultimately leading to an increased proliferation rate, a decrease in the percentage of SA-β-gal positive cells, upregulation of SIRT1 expression, and increased telomerase activity of the senescent ADSCs at the cellular level. Further recently it was observed that the ADSCs were treated with TC without induction of senescence, all the before said positives were observed. Moreover, a chemically induced liver fibrosis animal model was used to evaluate the functionality of these rescued cells in vivo. Liver dysfunction was established by injecting 200 mg/kg thioacetamide (TAA) intraperitoneally into Wistar rats every third day for 60 days. The experimental rats were separated into groups; normal group (rats without TAA induction), sham group (without ADSC transplantation), positive control group (transplanted with normal ADSCs); H2O2 group (transplanted with H2O2 -induced senescent ADSCs), H2O2+TC group (transplanted with ADSCs pretreated with H2O2 and then further treated with TC) and TC group (ADSC treated with TC without H2O2 treatment). In the transplantation group, 1 × 106 human ADSCs were introduced into each rat via direct liver injection. Based on the biochemical analysis and immunohistochemical staining results, it was determined that the therapeutic effects on liver fibrosis by the induced senescent ADSCs (H2O2 group) were not as significant as those exerted by the normal ADSCs (the positive control group). However, the H2O2+TC group showed significant reversal of liver damage when compared to the H2O2 group 1 week post-transplantation. Further ADSCs without H2O2 treatment but with just TC treatment performed much better than all the groups. These data confirmed that the TC treatment had the potential to improve the therapeutic effect of ADSCs. It is therefore suggested that TC has potential applications in maintaining stem cell quality and could possibly aid in the treatment of senescence-related disorders.

Keywords: senescence, SIRT1, adipose derived stem cells, liver fibrosis

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555 The Dynamics of a Droplet Spreading on a Steel Surface

Authors: Evgeniya Orlova, Dmitriy Feoktistov, Geniy Kuznetsov

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Spreading of a droplet over a solid substrate is a key phenomenon observed in the following engineering applications: thin film coating, oil extraction, inkjet printing, and spray cooling of heated surfaces. Droplet cooling systems are known to be more effective than film or rivulet cooling systems. It is caused by the greater evaporation surface area of droplets compared with the film of the same mass and wetting surface. And the greater surface area of droplets is connected with the curvature of the interface. Location of the droplets on the cooling surface influences on the heat transfer conditions. The close distance between the droplets provides intensive heat removal, but there is a possibility of their coalescence in the liquid film. The long distance leads to overheating of the local areas of the cooling surface and the occurrence of thermal stresses. To control the location of droplets is possible by changing the roughness, structure and chemical composition of the surface. Thus, control of spreading can be implemented. The most important characteristic of spreading of droplets on solid surfaces is a dynamic contact angle, which is a function of the contact line speed or capillary number. However, there is currently no universal equation, which would describe the relationship between these parameters. This paper presents the results of the experimental studies of water droplet spreading on metal substrates with different surface roughness. The effect of the droplet growth rate and the surface roughness on spreading characteristics was studied at low capillary numbers. The shadow method using high speed video cameras recording up to 10,000 frames per seconds was implemented. A droplet profile was analyzed by Axisymmetric Drop Shape Analyses techniques. According to change of the dynamic contact angle and the contact line speed three sequential spreading stages were observed: rapid increase in the dynamic contact angle; monotonous decrease in the contact angle and the contact line speed; and form of the equilibrium contact angle at constant contact line. At low droplet growth rate, the dynamic contact angle of the droplet spreading on the surfaces with the maximum roughness is found to increase throughout the spreading time. It is due to the fact that the friction force on such surfaces is significantly greater than the inertia force; and the contact line is pinned on microasperities of a relief. At high droplet growth rate the contact angle decreases during the second stage even on the surfaces with the maximum roughness, as in this case, the liquid does not fill the microcavities, and the droplet moves over the “air cushion”, i.e. the interface is a liquid/gas/solid system. Also at such growth rates pulsation of liquid flow was detected; and the droplet oscillates during the spreading. Thus, obtained results allow to conclude that it is possible to control spreading by using the surface roughness and the growth rate of droplets on surfaces as varied factors. Also, the research findings may be used for analyzing heat transfer in rivulet and drop cooling systems of high energy equipment.

Keywords: contact line speed, droplet growth rate, dynamic contact angle, shadow system, spreading

Procedia PDF Downloads 329
554 Using Convolutional Neural Networks to Distinguish Different Sign Language Alphanumerics

Authors: Stephen L. Green, Alexander N. Gorban, Ivan Y. Tyukin

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Within the past decade, using Convolutional Neural Networks (CNN)’s to create Deep Learning systems capable of translating Sign Language into text has been a breakthrough in breaking the communication barrier for deaf-mute people. Conventional research on this subject has been concerned with training the network to recognize the fingerspelling gestures of a given language and produce their corresponding alphanumerics. One of the problems with the current developing technology is that images are scarce, with little variations in the gestures being presented to the recognition program, often skewed towards single skin tones and hand sizes that makes a percentage of the population’s fingerspelling harder to detect. Along with this, current gesture detection programs are only trained on one finger spelling language despite there being one hundred and forty-two known variants so far. All of this presents a limitation for traditional exploitation for the state of current technologies such as CNN’s, due to their large number of required parameters. This work aims to present a technology that aims to resolve this issue by combining a pretrained legacy AI system for a generic object recognition task with a corrector method to uptrain the legacy network. This is a computationally efficient procedure that does not require large volumes of data even when covering a broad range of sign languages such as American Sign Language, British Sign Language and Chinese Sign Language (Pinyin). Implementing recent results on method concentration, namely the stochastic separation theorem, an AI system is supposed as an operate mapping an input present in the set of images u ∈ U to an output that exists in a set of predicted class labels q ∈ Q of the alphanumeric that q represents and the language it comes from. These inputs and outputs, along with the interval variables z ∈ Z represent the system’s current state which implies a mapping that assigns an element x ∈ ℝⁿ to the triple (u, z, q). As all xi are i.i.d vectors drawn from a product mean distribution, over a period of time the AI generates a large set of measurements xi called S that are grouped into two categories: the correct predictions M and the incorrect predictions Y. Once the network has made its predictions, a corrector can then be applied through centering S and Y by subtracting their means. The data is then regularized by applying the Kaiser rule to the resulting eigenmatrix and then whitened before being split into pairwise, positively correlated clusters. Each of these clusters produces a unique hyperplane and if any element x falls outside the region bounded by these lines then it is reported as an error. As a result of this methodology, a self-correcting recognition process is created that can identify fingerspelling from a variety of sign language and successfully identify the corresponding alphanumeric and what language the gesture originates from which no other neural network has been able to replicate.

Keywords: convolutional neural networks, deep learning, shallow correctors, sign language

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553 Recovering Trust in Institutions through Networked Governance: An Analytical Approach via the Study of the Provincial Government of Gipuzkoa

Authors: Xabier Barandiaran, Igone Guerra

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The economic and financial crisis that hit European countries in 2008 revealed the inability of governments to respond unilaterally to the so-called “wicked” problems that affect our societies. Closely linked to this, the increasing disaffection of citizens towards politics has resulted in growing distrust of the citizenry not only in the institutions in general but also in the political system, in particular. Precisely, these two factors provoked the action of the local government of Gipuzkoa (Basque Country) to move from old ways of “doing politics” to a new way of “thinking politics” based on a collaborative approach, in which innovative modes of public decision making are prominent. In this context, in 2015, the initiative Etorkizuna Eraikiz (Building the Future), a contemporary form of networked governance, was launched by the Provincial Government. The paper focuses on the Etorkizuna Eraikiz initiative, a sound commitment from a local government to build jointly with the citizens the future of the territory. This paper will present preliminary results obtained from three different experiences of co-creation developed within Etorkizuna Eraikiz in which the formulation of networked governance is a mandatory pre-requisite. These experiences show how the network building approach among the different agents of the territory as well as the co-creation of public policies is the cornerstone of this challenging mission. Through the analysis of the information and documentation gathered during the four years of Etorkizuna-Eraikiz, and, specifically by delving into the strategy promoted by the initiative, some emerging analytical conclusions resulting from the promotion of this collaborative culture will be presented. For example, some preliminary results have shown a significant positive relationship between shared leadership and the formulation of the public good. In the period 2016-2018, a total of 73 projects were launched and funding by the Provincial Government of Gipuzkoa within the Etorkizuna Eraikiz initiative, that indicates greater engagement of the citizenry in the process of policy-making and therefore improving, somehow, the quality of the public policies. These statements have been supported by the last survey about the perspectives of the citizens toward politics and policies. Some of the more prominent results show us that there is still a high level of distrust in Politics (78,9% of respondents) but a greater trust in institutions such the Political Government of Gipuzkoa (40,8% of respondents declared as “good” the performance of this provincial institution). Regarding the Etorkizuna Eraikiz Initiative, it is being more readily recognized by citizens over this period of time (25,4% of the respondents in June 2018 agreed to know about the initiative giving it a mark of 5,89 ) and thus build trust and a sense of ownership. Although, there is a clear requirement for further research on the linkages between collaborative governance and level of trust, the paper, based on these findings, will provide some managerial and theoretical implications for collaborative governance in the territory.

Keywords: network governance, collaborative governance, public sector innovation, citizen participation, trust

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552 The Role of Non-Governmental Organizations in Promoting Humanitarian Development: A Case Study in Saudi Arabia

Authors: Muamar Salameh, Rania Sinno

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Non-governmental organizations in Saudi Arabia play a vital role in promoting humanitarian development. Though this paper will emphasize this role and will provide a specific case study on the role of Prince Mohammad Bin Fahd Foundation for Humanitarian Development, yet many organizations do not provide transparent information for the accomplishments of the NGOs. This study will provide answers to the main research question regarding this role that NGOs play in promoting humanitarian development. The recent law regulating associations and foundations in Saudi Arabia was issued in December 2015 and went into effect March 2016. Any new association or foundation will need to follow these regulations. Though the registration, implementation, and workflow of the organizations still need major improvement and development, yet, the currently-registered organizations have several notable achievements. Most of these organizations adopt a centralized administration approach which in many cases still hinders progress and may be an obstacle in achieving and reaching a larger population of beneficiaries. A large portion of the existing organizations are charities, some of which have some sort of government affiliation. The laws and regulations limit registration of new organizations. Any violations to Islamic Sharia, contradictions to public order, breach to national unity, foreign and foreign-affiliation organizations prohibits any organization from registration. The lack of transparency in the operations and inner-working of NGOs in Saudi Arabia is apparent for the public. However, the regulations invoke full transparency with the governing ministry. This transparency should be available to the public and in specific to the target population that are eligible to benefit from the NGOs services. In this study, we will provide an extensive review of all related laws, regulations, policies and procedures related to all NGOs in the Eastern Province of Saudi Arabia. This review will include some examples of current NGOs, services and target population. The study will determine the main accomplishments of reputable NGOs that have impacted positively the Saudi communities. The results will highlight and concentrate on actions, services and accomplishments that achieve sustainable assistance in promoting humanitarian development and advance living conditions of target populations of the Saudi community. In particular, we will concentrate on a case study related to PMFHD; one of the largest foundations in the Eastern Province of Saudi Arabia. The authors have access to the data related to this foundation and have access to the foundation administration to gather, analyze and conclude the findings of this group. The study will also analyze whether the practices, budgets, services and annual accomplishments of the foundation have fulfilled the humanitarian role of the foundation while meeting the governmental requirements, with an analysis in the light of the new laws. The findings of the study show that great accomplishments for advancing and promoting humanitarian development in Saudi community and international communities have been achieved. Several examples will be included from several NGOs, with specific examples from PMFHD.

Keywords: development, foundation, humanitarian, non-governmental organization, Saudi Arabia

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551 Anti-Hyperglycemic Effects and Chemical Analysis of Allium sativum Bulbs Growing in Sudan

Authors: Ikram Mohamed Eltayeb Elsiddig, Yacouba Amina Djamila, Amna El Hassan Hamad

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Hyperglycemia and diabetes have been treated with several medicinal plants for a long time, meanwhile reduce associated side effects than the synthetic ones. Therefore, the search for more effective and safer anti-diabetic agents derived from plants has become an interest area of active research. A. sativum, belonging to the Liliaceae family is well known for its medicinal uses in African traditional medicine, it used for treating of many human diseases mainly diabetes, high cholesterol, and high blood pressure. The present study was carried out to investigate the anti-hyperglycemic effect of the extracts of A. sativum bulb growing in Sudan on glucose-loaded Wistar albino rats. A. sativum bulbs were collected from local vegetable market at Khourtoum/ Sudan in a fresh form, identified and authenticated by taxonomist, then dried, and extracted with solvents of increasing polarity: petroleum ether, chloroform, ethyl acetate and methanol by using Soxhlet apparatus. The effect of the extracts on glucose uptake was evaluated by using the isolated rats hemidiaphgrams after loading the fasting rats with glucose, and the anti-hyperglycemic effect was investigated on glucose-loaded Wistar albino rats. Their effects were compared to control rats administered with the vehicle and to a standard group administered with Metformin standard drug. The most active extract was analyzed chemically using GC-MS analysis compared to NIST library. The results showed significant anti-diabetic effect of extracts of A. sativum bulb growing in Sudan. Addition to the hypoglycemic activity of A. sativum extracts was found to be decreased with increase in the polarity of the extraction solvent; this may explain the less polarity of substance responsible for the activity and their concentration decreased with polarity increase. The petroleum ether extract possess anti-hyperglycemic activity more significant than the other extracts and the Metformin standard drug with p-value 0.000** of 400mg/kg at 1 hour, 2 hour and four hour; and p-value 0.019*, 0.015* and 0.010* of 200mg/kg at 1 hour, 2 hour and four hour respectively. The GC-MS analysis of petroleum ether extract, with highest anti -diabetes activity showed the presence of Methyl linolate (42.75%), Hexadecanoic acid, methyl ester (10.54%), Methyl α-linolenate (8.36%), Dotriacontane (6.83), Tetrapentacontane (6.33), Methyl 18-methylnonadecanoate (4.8), Phenol,2,2’-methylenebis[6-(1,1-dimethylethyl)-4-methyl] (3.25), Methyl 20-methyl-heneicosanoate (2.70), Pentatriacontane (2.13) and many other minor compounds. The most of these compounds are well known for their anti-diabetic activity. The study concluded that A. sativum bulbs extracts were found to enhanced the reuptake of glucose in the isolated rat hemidiaphragm and have antihyperglycemic effect when evaluated on glucose-loaded albino rats with petroleum ether extract activity more significant than the Metformin standard drug.

Keywords: Allium, anti-hyperglycemic, bulbs, sativum

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550 Thermal-Mechanical Analysis of a Bridge Deck to Determine Residual Weld Stresses

Authors: Evy Van Puymbroeck, Wim Nagy, Ken Schotte, Heng Fang, Hans De Backer

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The knowledge of residual stresses for welded bridge components is essential to determine the effect of the residual stresses on the fatigue life behavior. The residual stresses of an orthotropic bridge deck are determined by simulating the welding process with finite element modelling. The stiffener is placed on top of the deck plate before welding. A chained thermal-mechanical analysis is set up to determine the distribution of residual stresses for the bridge deck. First, a thermal analysis is used to determine the temperatures of the orthotropic deck for different time steps during the welding process. Twin wire submerged arc welding is used to construct the orthotropic plate. A double ellipsoidal volume heat source model is used to describe the heat flow through a material for a moving heat source. The heat input is used to determine the heat flux which is applied as a thermal load during the thermal analysis. The heat flux for each element is calculated for different time steps to simulate the passage of the welding torch with the considered welding speed. This results in a time dependent heat flux that is applied as a thermal loading. Thermal material behavior is specified by assigning the properties of the material in function of the high temperatures during welding. Isotropic hardening behavior is included in the model. The thermal analysis simulates the heat introduced in the two plates of the orthotropic deck and calculates the temperatures during the welding process. After the calculation of the temperatures introduced during the welding process in the thermal analysis, a subsequent mechanical analysis is performed. For the boundary conditions of the mechanical analysis, the actual welding conditions are considered. Before welding, the stiffener is connected to the deck plate by using tack welds. These tack welds are implemented in the model. The deck plate is allowed to expand freely in an upwards direction while it rests on a firm and flat surface. This behavior is modelled by using grounded springs. Furthermore, symmetry points and lines are used to prevent the model to move freely in other directions. In the thermal analysis, a mechanical material model is used. The calculated temperatures during the thermal analysis are introduced during the mechanical analysis as a time dependent load. The connection of the elements of the two plates in the fusion zone is realized with a glued connection which is activated when the welding temperature is reached. The mechanical analysis results in a distribution of the residual stresses. The distribution of the residual stresses of the orthotropic bridge deck is compared with results from literature. Literature proposes uniform tensile yield stresses in the weld while the finite element modelling showed tensile yield stresses at a short distance from the weld root or the weld toe. The chained thermal-mechanical analysis results in a distribution of residual weld stresses for an orthotropic bridge deck. In future research, the effect of these residual stresses on the fatigue life behavior of welded bridge components can be studied.

Keywords: finite element modelling, residual stresses, thermal-mechanical analysis, welding simulation

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549 Understanding the Experiences of School Teachers and Administrators Involved in a Multi-Sectoral Approach to the Creation of a Physical Literacy Enriched Community

Authors: M. Louise Humbert, Karen E. Chad, Natalie E. Houser, Marta E. Erlandson

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Physical literacy is the motivation, confidence, physical competence, knowledge, and understanding to value and takes responsibility for engagement in physical activities for life. In recent years, physical literacy has emerged as a determinant of health, promoting a positive lifelong physical activity trajectory. Physical literacy’s holistic approach and emphasis on the intrinsic valuation of movement provide an encouraging avenue for intervention among children to develop competent and confident movers. Although there is research on physical literacy interventions, no evidence exists on the outcomes of multi-sectoral interventions involving a combination of home, school, and community contexts. Since children interact with and in a wide range of contexts (home, school, community) daily, interventions designed to address a combination of these contexts are critical to the development of physical literacy. Working with school administrators and teachers, sports and recreation leaders, and community members, our team of university and community researchers conducted and evaluated one of the first multi-contextual and multi-sectoral physical literacy interventions in Canada. Schools played a critical role in this multi-sector intervention, and in this project, teachers and administrators focused their actions on developing physical literacy in students 10 to 14 years of age through the instruction of physical literacy-focused physical education lessons. Little is known about the experiences of educators when they work alongside an array of community representatives to develop physical literacy in school-aged children. Given the uniqueness of this intervention, we sought to answer the question, ‘What were the experiences of school-based educators involved in a multi-sectoral partnership focused on creating a physical literacy enriched community intervention?’ A thematic analysis approach was used to analyze data collected from interviews with educators and administrators, informal conversations, documents, and observations at workshops and meetings. Results indicated that schools and educators played the largest role in this multi-sector intervention. Educators initially reported a limited understanding of physical literacy and expressed a need for resources linked to the physical education curriculum. Some anxiety was expressed by the teachers as their students were measured, and educators noted they wanted to increase their understanding and become more involved in the assessment of physical literacy. Teachers reported that the intervention’s focus on physical literacy positively impacted the scheduling and their instruction of physical education. Administrators shared their desire for school and division-level actions targeting physical literacy development like the current focus on numeracy and literacy, treaty education, and safe schools. As this was one of the first multi-contextual and multi-sectoral physical literacy interventions, it was important to document creation and delivery experiences to encourage future growth in the area and develop suggested best practices.

Keywords: physical literacy, multi sector intervention, physical education, teachers

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548 Ammonia Bunkering Spill Scenarios: Modelling Plume’s Behaviour and Potential to Trigger Harmful Algal Blooms in the Singapore Straits

Authors: Bryan Low

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In the coming decades, the global maritime industry will face a most formidable environmental challenge -achieving net zero carbon emissions by 2050. To meet this target, the Maritime Port Authority of Singapore (MPA) has worked to establish green shipping and digital corridors with ports of several other countries around the world where ships will use low-carbon alternative fuels such as ammonia for power generation. While this paradigm shift to the bunkering of greener fuels is encouraging, fuels like ammonia will also introduce a new and unique type of environmental risk in the unlikely scenario of a spill. While numerous modelling studies have been conducted for oil spills and their associated environmental impact on coastal and marine ecosystems, ammonia spills are comparatively less well understood. For example, there is a knowledge gap regarding how the complex hydrodynamic conditions of the Singapore Straits may influence the dispersion of a hypothetical ammonia plume, which has different physical and chemical properties compared to an oil slick. Chemically, ammonia can be absorbed by phytoplankton, thus altering the balance of the marine nitrogen cycle. Biologically, ammonia generally serves the role of a nutrient in coastal ecosystems at lower concentrations. However, at higher concentrations, it has been found to be toxic to many local species. It may also have the potential to trigger eutrophication and harmful algal blooms (HABs) in coastal waters, depending on local hydrodynamic conditions. Thus, the key objective of this research paper is to support the development of a model-based forecasting system that can predict ammonia plume behaviour in coastal waters, given prevailing hydrodynamic conditions and their environmental impact. This will be essential as ammonia bunkering becomes more commonplace in Singapore’s ports and around the world. Specifically, this system must be able to assess the HAB-triggering potential of an ammonia plume, as well as its lethal and sub-lethal toxic effects on local species. This will allow the relevant authorities to better plan risk mitigation measures or choose a time window with the ideal hydrodynamic conditions to conduct ammonia bunkering operations with minimal risk. In this paper, we present the first part of such a forecasting system: a jointly coupled hydrodynamic-water quality model that can capture how advection-diffusion processes driven by ocean currents influence plume behaviour and how the plume interacts with the marine nitrogen cycle. The model is then applied to various ammonia spill scenarios where the results are discussed in the context of current ammonia toxicity guidelines, impact on local ecosystems, and mitigation measures for future bunkering operations conducted in the Singapore Straits.

Keywords: ammonia bunkering, forecasting, harmful algal blooms, hydrodynamics, marine nitrogen cycle, oceanography, water quality modeling

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547 Solids and Nutrient Loads Exported by Preserved and Impacted Low-Order Streams: A Comparison among Water Bodies in Different Latitudes in Brazil

Authors: Nicolas R. Finkler, Wesley A. Saltarelli, Taison A. Bortolin, Vania E. Schneider, Davi G. F. Cunha

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Estimating the relative contribution of nonpoint or point sources of pollution in low-orders streams is an important tool for the water resources management. The location of headwaters in areas with anthropogenic impacts from urbanization and agriculture is a common scenario in developing countries. This condition can lead to conflicts among different water users and compromise ecosystem services. Water pollution also contributes to exporting organic loads to downstream areas, including higher order rivers. The purpose of this research is to preliminarily assess nutrients and solids loads exported by water bodies located in watersheds with different types of land uses in São Carlos - SP (Latitude. -22.0087; Longitude. -47.8909) and Caxias do Sul - RS (Latitude. -29.1634, Longitude. -51.1796), Brazil, using regression analysis. The variables analyzed in this study were Total Kjeldahl Nitrogen (TKN), Nitrate (NO3-), Total Phosphorus (TP) and Total Suspended Solids (TSS). Data were obtained in October and December 2015 for São Carlos (SC) and in November 2012 and March 2013 for Caxias do Sul (CXS). Such periods had similar weather patterns regarding precipitation and temperature. Altogether, 11 sites were divided into two groups, some classified as more pristine (SC1, SC4, SC5, SC6 and CXS2), with predominance of native forest; and others considered as impacted (SC2, SC3, CXS1, CXS3, CXS4 and CXS5), presenting larger urban and/or agricultural areas. Previous linear regression was applied for data on flow and drainage area of each site (R² = 0.9741), suggesting that the loads to be assessed had a significant relationship with the drainage areas. Thereafter, regression analysis was conducted between the drainage areas and the total loads for the two land use groups. The R² values were 0.070, 0.830, 0.752 e 0.455 respectively for SST, TKN, NO3- and TP loads in the more preserved areas, suggesting that the loads generated by runoff are significant in these locations. However, the respective R² values for sites located in impacted areas were respectively 0.488, 0.054, 0.519 e 0.059 for SST, TKN, NO3- and P loads, indicating a less important relationship between total loads and runoff as compared to the previous scenario. This study suggests three possible conclusions that will be further explored in the full-text article, with more sampling sites and periods: a) In preserved areas, nonpoint sources of pollution are more significant in determining water quality in relation to the studied variables; b) The nutrient (TKN and P) loads in impacted areas may be associated with point sources such as domestic wastewater discharges with inadequate treatment levels; and c) The presence of NO3- in impacted areas can be associated to the runoff, particularly in agricultural areas, where the application of fertilizers is common at certain times of the year.

Keywords: land use, linear regression, point and non-point pollution sources, streams, water resources management

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546 Graphene Metamaterials Supported Tunable Terahertz Fano Resonance

Authors: Xiaoyong He

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The manipulation of THz waves is still a challenging task due to lack of natural materials interacted with it strongly. Designed by tailoring the characters of unit cells (meta-molecules), the advance of metamaterials (MMs) may solve this problem. However, because of Ohmic and radiation losses, the performance of MMs devices is subjected to the dissipation and low quality factor (Q-factor). This dilemma may be circumvented by Fano resonance, which arises from the destructive interference between a bright continuum mode and dark discrete mode (or a narrow resonance). Different from symmetric Lorentz spectral curve, Fano resonance indicates a distinct asymmetric line-shape, ultrahigh quality factor, steep variations in spectrum curves. Fano resonance is usually realized through symmetry breaking. However, if concentric double rings (DR) are placed closely to each other, the near-field coupling between them gives rise to two hybridized modes (bright and narrowband dark modes) because of the local asymmetry, resulting into the characteristic Fano line shape. Furthermore, from the practical viewpoint, it is highly desirable requirement that to achieve the modulation of Fano spectral curves conveniently, which is an important and interesting research topics. For current Fano systems, the tunable spectral curves can be realized by adjusting the geometrical structural parameters or magnetic fields biased the ferrite-based structure. But due to limited dispersion properties of active materials, it is still a tough work to tailor Fano resonance conveniently with the fixed structural parameters. With the favorable properties of extreme confinement and high tunability, graphene is a strong candidate to achieve this goal. The DR-structure possesses the excitation of so-called “trapped modes,” with the merits of simple structure and high quality of resonances in thin structures. By depositing graphene circular DR on the SiO2/Si/ polymer substrate, the tunable Fano resonance has been theoretically investigated in the terahertz regime, including the effects of graphene Fermi level, structural parameters and operation frequency. The results manifest that the obvious Fano peak can be efficiently modulated because of the strong coupling between incident waves and graphene ribbons. As Fermi level increases, the peak amplitude of Fano curve increases, and the resonant peak position shifts to high frequency. The amplitude modulation depth of Fano curves is about 30% if Fermi level changes in the scope of 0.1-1.0 eV. The optimum gap distance between DR is about 8-12 μm, where the value of figure of merit shows a peak. As the graphene ribbon width increases, the Fano spectral curves become broad, and the resonant peak denotes blue shift. The results are very helpful to develop novel graphene plasmonic devices, e.g. sensors and modulators.

Keywords: graphene, metamaterials, terahertz, tunable

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545 Consensus, Federalism and Inter-State Water Disputes in India

Authors: Amrisha Pandey

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Indian constitution has distributed the powers to govern and legislate between the centre and the state governments based on the list of subject-matter provided in the seventh schedule. By that schedule, the states are authorized to regulate the water resource within their territory. However, the centre/union government is authorized to regulate the inter-state water disputes. The powers entrusted to the union government mainly deals with the sharing of river water which flows through the territory of two or more states. For that purpose, a provision enumerated in Article 262 of the Constitution of India which empowers the parliament to resolve any such inter-state river water dispute. Therefore, the parliament has enacted the - ‘Inter-State River Water Dispute Tribunal, Act’, which allows the central/union government to constitute the tribunal for the adjudication of the disputes and expressly bars the jurisdiction of the judiciary in the concerned matter. This arrangement was intended to resolve the dispute using political or diplomatic means, without deliberately interfering with the sovereign power of the states to govern the water resource. The situation in present context is complicated and sensitive. Due to the change in climatic conditions; increasing demand for the limited resource; and the advanced understanding of the freshwater cycle, which is missing from the existing legal regime. The obsolete legal and political tools, the existing legislative mechanism and the institutional units do not seem to accommodate the rising challenge to regulate the resource. Therefore, resulting in the rise of the politicization of the inter-state water disputes. Against this background, this paper will investigate the inter-state river water dispute in India and will critically analyze the ability of the existing constitutional, and institutional units involved in the task. Moreover, the competence of the tribunal as the adjudicating body in present context will be analyzed using the long ongoing inter-state water dispute in India – The Cauvery Water Dispute, as the case study. To conduct the task undertaken in this paper the doctrinal methodology of the research is adopted. The disputes will also be investigated through the lens of sovereignty, which is accorded to the states using the theory of ‘separation of power’ and the ‘grant of internal sovereignty’, to its federal units of governance. The issue of sovereignty in this paper is discussed in two ways: 1) as the responsibility of the state - to govern the resource; and 2) as the obligation of the state - to govern the resource, arising from the sovereign power of the state. Furthermore, the duality of the sovereign power coexists in this analysis; the overall sovereign authority of the nation-state, and the internal sovereignty of the states as its federal units of governance. As a result, this investigation will propose institutional, legislative and judicial reforms. Additionally, it will suggest certain amendments to the existing constitutional provisions in order to avoid the contradictions in their scope and meaning in the light of the advanced hydrological understanding.

Keywords: constitution of India, federalism, inter-state river water dispute tribunal of India, sovereignty

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544 Exploring the Use of Augmented Reality for Laboratory Lectures in Distance Learning

Authors: Michele Gattullo, Vito M. Manghisi, Alessandro Evangelista, Enricoandrea Laviola

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In this work, we explored the use of Augmented Reality (AR) to support students in laboratory lectures in Distance Learning (DL), designing an application that proved to be ready for use next semester. AR could help students in the understanding of complex concepts as well as increase their motivation in the learning process. However, despite many prototypes in the literature, it is still less used in schools and universities. This is mainly due to the perceived limited advantages to the investment costs, especially regarding changes needed in the teaching modalities. However, with the spread of epidemiological emergency due to SARS-CoV-2, schools and universities were forced to a very rapid redefinition of consolidated processes towards forms of Distance Learning. Despite its many advantages, it suffers from the impossibility to carry out practical activities that are of crucial importance in STEM ("Science, Technology, Engineering e Math") didactics. In this context, AR perceived advantages increased a lot since teachers are more prepared for new teaching modalities, exploiting AR that allows students to carry on practical activities on their own instead of being physically present in laboratories. In this work, we designed an AR application for the support of engineering students in the understanding of assembly drawings of complex machines. Traditionally, this skill is acquired in the first years of the bachelor's degree in industrial engineering, through laboratory activities where the teacher shows the corresponding components (e.g., bearings, screws, shafts) in a real machine and their representation in the assembly drawing. This research aims to explore the effectiveness of AR to allow students to acquire this skill on their own without physically being in the laboratory. In a preliminary phase, we interviewed students to understand the main issues in the learning of this subject. This survey revealed that students had difficulty identifying machine components in an assembly drawing, matching between the 2D representation of a component and its real shape, and understanding the functionality of a component within the machine. We developed a mobile application using Unity3D, aiming to solve the mentioned issues. We designed the application in collaboration with the course professors. Natural feature tracking was used to associate the 2D printed assembly drawing with the corresponding 3D virtual model. The application can be displayed on students’ tablets or smartphones. Users could interact with selecting a component from a part list on the device. Then, 3D representations of components appear on the printed drawing, coupled with 3D virtual labels for their location and identification. Users could also interact with watching a 3D animation to learn how components are assembled. Students evaluated the application through a questionnaire based on the System Usability Scale (SUS). The survey was provided to 15 students selected among those we participated in the preliminary interview. The mean SUS score was 83 (SD 12.9) over a maximum of 100, allowing teachers to use the AR application in their courses. Another important finding is that almost all the students revealed that this application would provide significant power for comprehension on their own.

Keywords: augmented reality, distance learning, STEM didactics, technology in education

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543 Effect of the Polymer Modification on the Cytocompatibility of Human and Rat Cells

Authors: N. Slepickova Kasalkova, P. Slepicka, L. Bacakova, V. Svorcik

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Tissue engineering includes combination of materials and techniques used for the improvement, repair or replacement of the tissue. Scaffolds, permanent or temporally material, are used as support for the creation of the "new cell structures". For this important component (scaffold), a variety of materials can be used. The advantage of some polymeric materials is their cytocompatibility and possibility of biodegradation. Poly(L-lactic acid) (PLLA) is a biodegradable,  semi-crystalline thermoplastic polymer. PLLA can be fully degraded into H2O and CO2. In this experiment, the effect of the surface modification of biodegradable polymer (performed by plasma treatment) on the various cell types was studied. The surface parameters and changes of the physicochemical properties of modified PLLA substrates were studied by different methods. Surface wettability was determined by goniometry, surface morphology and roughness study were performed with atomic force microscopy and chemical composition was determined using photoelectron spectroscopy. The physicochemical properties were studied in relation to cytocompatibility of human osteoblast (MG 63 cells), rat vascular smooth muscle cells (VSMC), and human stem cells (ASC) of the adipose tissue in vitro. A fluorescence microscopy was chosen to study and compare cell-material interaction. Important parameters of the cytocompatibility like adhesion, proliferation, viability, shape, spreading of the cells were evaluated. It was found that the modification leads to the change of the surface wettability depending on the time of modification. Short time of exposition (10-120 s) can reduce the wettability of the aged samples, exposition longer than 150 s causes to increase of contact angle of the aged PLLA. The surface morphology is significantly influenced by duration of modification, too. The plasma treatment involves the formation of the crystallites, whose number increases with increasing time of modification. On the basis of physicochemical properties evaluation, the cells were cultivated on the selected samples. Cell-material interactions are strongly affected by material chemical structure and surface morphology. It was proved that the plasma treatment of PLLA has a positive effect on the adhesion, spreading, homogeneity of distribution and viability of all cultivated cells. This effect was even more apparent for the VSMCs and ASCs which homogeneously covered almost the whole surface of the substrate after 7 days of cultivation. The viability of these cells was high (more than 98% for VSMCs, 89-96% for ASCs). This experiment is one part of the basic research, which aims to easily create scaffolds for tissue engineering with subsequent use of stem cells and their subsequent "reorientation" towards the bone cells or smooth muscle cells.

Keywords: poly(L-lactic acid), plasma treatment, surface characterization, cytocompatibility, human osteoblast, rat vascular smooth muscle cells, human stem cells

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542 Role of Empirical Evidence in Law-Making: Case Study from India

Authors: Kaushiki Sanyal, Rajesh Chakrabarti

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In India, on average, about 60 Bills are passed every year in both Houses of Parliament – Lok Sabha and Rajya Sabha (calculated from information on websites of both Houses). These are debated in both Lok Sabha (House of Commons) and Rajya Sabha (Council of States) before they are passed. However, lawmakers rarely use empirical evidence to make a case for a law. Most of the time, they support a law on the basis of anecdote, intuition, and common sense. While these do play a role in law-making, without the necessary empirical evidence, laws often fail to achieve their desired results. The quality of legislative debates is an indicator of the efficacy of the legislative process through which a Bill is enacted. However, the study of legislative debates has not received much attention either in India or worldwide due to the difficulty of objectively measuring the quality of a debate. Broadly, three approaches have emerged in the study of legislative debates. The rational-choice or formal approach shows that speeches vary based on different institutional arrangements, intra-party politics, and the political culture of a country. The discourse approach focuses on the underlying rules and conventions and how they impact the content of the debates. The deliberative approach posits that legislative speech can be reasoned, respectful, and informed. This paper aims to (a) develop a framework to judge the quality of debates by using the deliberative approach; (b) examine the legislative debates of three Bills passed in different periods as a demonstration of the framework, and (c) examine the broader structural issues that disincentive MPs from scrutinizing Bills. The framework would include qualitative and quantitative indicators to judge a debate. The idea is that the framework would provide useful insights into the legislators’ knowledge of the subject, the depth of their scrutiny of Bills, and their inclination toward evidence-based research. The three Bills that the paper plans to examine are as follows: 1. The Narcotics Drugs and Psychotropic Substances Act, 1985: This act was passed to curb drug trafficking and abuse. However, it mostly failed to fulfill its purpose. Consequently, it was amended thrice but without much impact on the ground. 2. The Criminal Laws (Amendment) Act, 2013: This act amended the Indian Penal Code to add a section on human trafficking. The purpose was to curb trafficking and penalise traffickers, pimps, and middlemen. However, the crime rate remains high while the conviction rate is low. 3. The Surrogacy (Regulation) Act, 2021: This act bans commercial surrogacy allowing only relatives to act as surrogates as long as there is no monetary payment. Experts fear that instead of preventing commercial surrogacy, it would drive the activity underground. The consequences would be borne by the surrogate, who would not be protected by law. The purpose of the paper is to objectively analyse the quality of parliamentary debates, get insights into how MPs understand the evidence and deliberate on steps to incentivise them to use empirical evidence.

Keywords: legislature, debates, empirical, India

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541 Digital Technology Relevance in Archival and Digitising Practices in the Republic of South Africa

Authors: Tashinga Matindike

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By means of definition, digital artworks encompass an array of artistic productions that are expressed in a technological form as an essential part of a creative process. Examples include illustrations, photos, videos, sculptures, and installations. Within the context of the visual arts, the process of repatriation involves the return of once-appropriated goods. Archiving denotes the preservation of a commodity for storage purposes in order to nurture its continuity. The aforementioned definitions form the foundation of the academic framework and premise of the argument, which is outlined in this paper. This paper aims to define, discuss and decipher the complexities involved in digitising artworks, whilst explaining the benefits of the process, particularly within the South African context, which is rich in tangible and intangible traditional cultural material, objects, and performances. With the internet having been introduced to the African Continent in the early 1990s, this new form of technology, in its own right, initiated a high degree of efficiency, which also resulted in the progressive transformation of computer-generated visual output. Subsequently, this caused a revolutionary influence on the manner in which technological software was developed and uterlised in art-making. Digital technology and the digitisation of creative processes then opened up new avenues of collating and recording information. One of the first visual artists to make use of digital technology software in his creative productions was United States-based artist John Whitney. His inventive work contributed greatly to the onset and development of digital animation. Comparable by technique and originality, South African contemporary visual artists who make digital artworks, both locally and internationally, include David Goldblatt, Katherine Bull, Fritha Langerman, David Masoga, Zinhle Sethebe, Alicia Mcfadzean, Ivan Van Der Walt, Siobhan Twomey, and Fhatuwani Mukheli. In conclusion, the main objective of this paper is to address the following questions: In which ways has the South African art community of visual artists made use of and benefited from technology, in its digital form, as a means to further advance creativity? What are the positive changes that have resulted in art production in South Africa since the onset and use of digital technological software? How has digitisation changed the manner in which we record, interpret, and archive both written and visual information? What is the role of South African art institutions in the development of digital technology and its use in the field of visual art. What role does digitisation play in the process of the repatriation of artworks and artefacts. The methodology in terms of the research process of this paper takes on a multifacted form, inclusive of data analysis of information attained by means of qualitative and quantitative approaches.

Keywords: digital art, digitisation, technology, archiving, transformation and repatriation

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540 Developing Granular Sludge and Maintaining High Nitrite Accumulation for Anammox to Treat Municipal Wastewater High-efficiently in a Flexible Two-stage Process

Authors: Zhihao Peng, Qiong Zhang, Xiyao Li, Yongzhen Peng

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Nowadays, conventional nitrogen removal process (nitrification and denitrification) was adopted in most wastewater treatment plants, but many problems have occurred, such as: high aeration energy consumption, extra carbon sources dosage and high sludge treatment costs. The emergence of anammox has bring about the great revolution to the nitrogen removal technology, and only the ammonia and nitrite were required to remove nitrogen autotrophically, no demand for aeration and sludge treatment. However, there existed many challenges in anammox applications: difficulty of biomass retention, insufficiency of nitrite substrate, damage from complex organic etc. Much effort was put into the research in overcoming the above challenges, and the payment was rewarded. It was also imperative to establish an innovative process that can settle the above problems synchronously, after all any obstacle above mentioned can cause the collapse of anammox system. Therefore, in this study, a two-stage process was established that the sequencing batch reactor (SBR) and upflow anaerobic sludge blanket (UASB) were used in the pre-stage and post-stage, respectively. The domestic wastewater entered into the SBR first and went through anaerobic/aerobic/anoxic (An/O/A) mode, and the draining at the aerobic end of SBR was mixed with domestic wastewater, the mixture then entering to the UASB. In the long term, organic and nitrogen removal performance was evaluated. All along the operation, most COD was removed in pre-stage (COD removal efficiency > 64.1%), including some macromolecular organic matter, like: tryptophan, tyrosinase and fulvic acid, which could weaken the damage of organic matter to anammox. And the An/O/A operating mode of SBR was beneficial to the achievement and maintenance of partial nitrification (PN). Hence, sufficient and steady nitrite supply was another favorable condition to anammox enhancement. Besides, the flexible mixing ratio helped to gain a substrate ratio appropriate to anammox (1.32-1.46), which further enhance the anammox. Further, the UASB was used and gas recirculation strategy was adopted in the post-stage, aiming to achieve granulation by the selection pressure. As expected, the granules formed rapidly during 38 days, which increased from 153.3 to 354.3 μm. Based on bioactivity and gene measurement, the anammox metabolism and abundance level rose evidently, by 2.35 mgN/gVss·h and 5.3 x109. The anammox bacteria mainly distributed in the large granules (>1000 μm), while the biomass in the flocs (<200 μm) and microgranules (200-500 μm) barely displayed anammox bioactivity. Enhanced anammox promoted the advanced autotrophic nitrogen removal, which increased from 71.9% to 93.4%, even when the temperature was only 12.9 ℃. Therefore, it was feasible to enhance anammox in the multiple favorable conditions created, and the strategy extended the application of anammox to the full-scale mainstream, enhanced the understanding of anammox in the aspects of culturing conditions.

Keywords: anammox, granules, nitrite accumulation, nitrogen removal efficiency

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539 Concussion: Clinical and Vocational Outcomes from Sport Related Mild Traumatic Brain Injury

Authors: Jack Nash, Chris Simpson, Holly Hurn, Ronel Terblanche, Alan Mistlin

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There is an increasing incidence of mild traumatic brain injury (mTBI) cases throughout sport and with this, a growing interest from governing bodies to ensure these are managed appropriately and player welfare is prioritised. The Berlin consensus statement on concussion in sport recommends a multidisciplinary approach when managing those patients who do not have full resolution of mTBI symptoms. There are as of yet no standardised guideline to follow in the treatment of complex cases mTBI in athletes. The aim of this project was to analyse the outcomes, both clinical and vocational, of all patients admitted to the mild Traumatic Brain Injury (mTBI) service at the UK’s Defence Military Rehabilitation Centre Headley Court between 1st June 2008 and 1st February 2017, as a result of a sport induced injury, and evaluate potential predictive indicators of outcome. Patients were identified from a database maintained by the mTBI service. Clinical and occupational outcomes were ascertained from medical and occupational employment records, recorded prospectively, at time of discharge from the mTBI service. Outcomes were graded based on the vocational independence scale (VIS) and clinical documentation at discharge. Predictive indicators including referral time, age at time of injury, previous mental health diagnosis and a financial claim in place at time of entry to service were assessed using logistic regression. 45 Patients were treated for sport-related mTBI during this time frame. Clinically 96% of patients had full resolution of their mTBI symptoms after input from the mTBI service. 51% of patients returned to work at their previous vocational level, 4% had ongoing mTBI symptoms, 22% had ongoing physical rehabilitation needs, 11% required mental health input and 11% required further vestibular rehabilitation. Neither age, time to referral, pre-existing mental health condition nor compensation seeking had a significant impact on either vocational or clinical outcome in this population. The vast majority of patients reviewed in the mTBI clinic had persistent symptoms which could not be managed in primary care. A consultant-led, multidisciplinary approach to the diagnosis and management of mTBI has resulted in excellent clinical outcomes in these complex cases. High levels of symptom resolution suggest that this referral and treatment pathway is successful and is a model which could be replicated in other organisations with consultant led input. Further understanding of both predictive and individual factors would allow clinicians to focus treatments on those who are most likely to develop long-term complications following mTBI. A consultant-led, multidisciplinary service ensures a large number of patients will have complete resolution of mTBI symptoms after sport-related mTBI. Further research is now required to ascertain the key predictive indicators of outcome following sport-related mTBI.

Keywords: brain injury, concussion, neurology, rehabilitation, sports injury

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538 MTT Assay-Guided Isolation of a Cytotoxic Lead from Hedyotis umbellata and Its Mechanism of Action against Non-Small Cell Lung Cancer A549 Cells

Authors: Kirti Hira, A. Sajeli Begum, S. Mahibalan, Poorna Chandra Rao

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Introduction: Cancer is one of the leading causes of death worldwide. Although existing therapy effectively kills cancer cells, they do affect normal growing cells leading to many undesirable side effects. Hence there is need to develop effective as well as safe drug molecules to combat cancer, which is possible through phyto-research. The currently available plant-derived blockbuster drugs are the example for this. In view of this, an investigation was done to identify cytotoxic lead molecules from Hedyotis umbellata (Family Rubiaceae), a widely distributed weed in India. Materials and Methods: The methanolic extract of the whole plant of H. umbellata (MHU), prepared through Soxhlet extraction method was further fractionated with diethyl ether and n-butanol, successively. MHU, ether fraction (EMHU) and butanol fraction (BMHU) were lyophilized and were tested for the cytotoxic effect using 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay against non-small cell lung cancer (NSCLC) A549 cell lines. The potentially active EMHU was subjected to chromatographic purification using normal-phase silica columns, in order to isolate the responsible bioactive compounds. The isolated pure compounds were tested for their cytotoxic effect by MTT assay against A549 cells. Compound-3, which was found to be most active, was characterized using IR, 1H- and 13C-NMR and MS analysis. The study was further extended to decipher the mechanism of action of cytotoxicity of compound-3 against A549 cells through various in vitro cellular models. Cell cycle analysis was done using flow cytometry following PI (Propidium Iodide) staining. Protein analysis was done using Western blot technique. Results: Among MHU, EMHU, and BMHU, the non-polar fraction EMHU demonstrated a significant dose-dependent cytotoxic effect with IC50 of 67.7μg/ml. Chromatography of EMHU yielded seven compounds. MTT assay of isolated compounds explored compound-3 as potentially active one, which inhibited the growth of A549 cells with IC50value of 14.2μM. Further, compound-3 was identified as cedrelopsin, a coumarin derivative having molecular weight of 260. Results of in vitro mechanistic studies explained that cedrelopsin induced cell cycle arrest at G2/M phase and down-regulated the expression of G2/M regulatory proteins such as cyclin B1, cdc2, and cdc25C, dose dependently. This is the first report that explores the cytotoxic mechanism of cedrelopsin. Conclusion: Thus a potential small lead molecule, cedrelopsin isolated from H. umbellata, showing antiproliferative effect mediated by G2/M arrest in A549 cells was discovered. The effect of cedrelopsin against other cancer cell lines followed by in vivo studies can be performed in future to develop a new drug candidate.

Keywords: A549, cedrelopsin, G2/M phase, Hedyotis umbellata

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