Search results for: culture and civilization
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 3726

Search results for: culture and civilization

2856 The Regional Expression of New Rural Dwellings Design in Linhai, Zhejiang: A Case of New Rural Dwellings Design in Badie Village

Authors: Fan Zhang

Abstract:

In the process of urbanization in China, the new rural construction is in the ascendant, which is becoming more and more popular. Under the driving effect of rural urbanization, the house pattern and tectonic methods of traditional vernacular houses have shown great differences from the family structure and values of contemporary peasant families. Therefore, it is particularly important to find a prototype, form and strategy, to make a balance between the traditional memory and modern functional requirements. In order for research to combine the regional culture with modern life, under the situation of the current batch production of new rural residence, Badie village, in Linhai, Zhejiang province, is taken as the case. This paper aims to put forward a prototype which can not only meet the demand of modern life but also ensure the continuation of traditional culture and historical context for the new rural dwellings design. This research not only helps to extend the local context in the construction of the new site but also contributes to the fusion of old and new rural dwellings in the old site construction. Through the study and research of this case, the research methodology and results can be drawn as reference for the new rural construction in other areas.

Keywords: badie village, design strategy, new rural dwellings, regional context, regional expression

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2855 Homosexuality in Burundi and Homosexuals Rights

Authors: Ciza Didier

Abstract:

By definition, homosexuality designates the sexual or amorous attraction towards a person of the same sex or of the same gender as one's own. The Burundi country has superficially 27834km2 with 13 millions of population. There are groups of certain people assuming that they are homosexual and that they want to claim their rights. Burundian homosexuals often organise seminars in the premises of the National Health Security Agency (NHSA) located at Kigobe quarter, in Bujumbura, this is the place where they meet to try to exchange and create their association for claim their rights. There are 2 categories of homosexuals: - gays: homosexuality between men (male sex) - lesbians: homosexuality between women (female sex) In the gay couple, there is one who behaves like a woman and often wears feminine styles while the other always remains like a man and always wears masculine styles. In the lesbian couple, there is one who behaves like a man and wears men's styles while the other remains as she is like a woman. In general, Burundian society is against homosexuality. Our society sees them as pariahs carrying a curse. According to Burundian culture and customs, homosexuality is satanic, therefore it is a great sin. In April 2011, Burundian President Pierre Nkurunziza signed a law criminalizing homosexual acts and providing for a sentence of three months to two years in prison, as well as a fine of BIF 50,000 to BIF 100,000 for any homosexual behavior. The investigation recently done shows that out of 300 people questioned, 299 were against homosexuality saying that it is against Burundian culture and 1 was for homosexuality. All Burundians are not against homosexuality. Their country must therefore take into consideration the small party of people who are for homosexuality. Homosexuals, too, need to live like others.

Keywords: homosexuality, lesbian, gay, law

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2854 Derivation of Human NK Cells from T Cell-Derived Induced Pluripotent Stem Cells Using Xenogeneic Serum-Free and Feeder Cell-Free Culture System

Authors: Aliya Sekenova, Vyacheslav Ogay

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The derivation of human induced pluripotent stem cells (iPSCs) from somatic cells by direct reprogramming opens wide perspectives in the regenerative medicine. It means the possibility to develop the personal and, consequently, any immunologically compatible cells for applications in cell-based therapy. The purpose of our study was to develop the technology for the production of NK cells from T cell-derived induced pluripotent stem cells (TiPSCs) for subsequent application in adoptive cancer immunotherapy. Methods: In this study iPSCs were derived from peripheral blood T cells using Sendai virus vectors expressing Oct4, Sox2, Klf4 and c-Myc. Pluripotent characteristics of TiPSCs were examined and confirmed with alkaline phosphatase staining, immunocytochemistry and RT-PCR analysis. For NK cell differentiation, embryoid bodies (EB) formed from (TiPSCs) were cultured in xenogeneic serum-free medium containing human serum, IL-3, IL-7, IL-15, SCF, FLT3L without using M210-B4 and AFT-024 stromal feeder cells. After differentiation, NK cells were characterized with immunofluorescence analysis, flow cytometry and cytotoxicity assay. Results: Here, we for the first time demonstrate that TiPSCs can effectively differentiate into functionally active NK cells without M210-B4 and AFT-024 xenogeneic stroma cells. Immunofluorescence and flow cytometry analysis showed that EB-derived cells can differentiate into a homogeneous population of NK cell expressing high levels of CD56, CD45 and CD16 specific markers. Moreover, these cells significantly express killing activation receptors such as NKp44 and NKp46. In the comparative analysis, we observed that NK cells derived using feeder-free culture system have more high killing activity against K-562 tumor cells, than NK cells derived by feeder-dependent method. Thus, we think that our obtained data will be useful for the development of large-scale production of NK cells for translation into cancer immunotherapy.

Keywords: induced pluripotent stem cells, NK cells, T cells, cell diffentiation, feeder cell-free culture system

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2853 UV-Reactive Electrospinning: Preparation, Characterization and Cell Culture Applications of Nanofiber Scaffolds Containing Keratin

Authors: Duygu Yüksel Deniz, Memet Vezir Kahraman, Serap Erdem Kuruca, Mediha Süleymanoğlu

Abstract:

Our first aim was to synthesize Hydroxy Apatite (HAP) and then modify its surface by adding 4-Vinylbenzene boronic acid (4-VBBA). The characterization was done by FT-IR. By adding Polyvinyl alcohol (PVA) to 4- VBBA-HAP, we obtained a suitable electrospinning solution. PVA solution which was also modified by using alkoxy silanes, in order to prevent the scaffolds from being damaged by aqueous cell medium, was added. Keratin was dissolved and then added into the electrospinning solution. Keratin containing 4-VBBA- HAP/PVA composite was used to fabricate nanofiber scaffolds with the simultaneous UV-reactive electrospinning technique. The structural characterization was done by FT-IR. Thermal gravimetric analysis was also performed by using TGA. The morphological characterization was determined by SEM analyses. Our second aim was to create a scaffold where cells could grow. With this purpose, suitable nanofibers were choosen according to their SEM analysis. Keratin containing nanofibers were seeded with 3T3, ECV and SAOS cells and their cytotoxicity and cell proliferation were investigated by using MTT assay. After cell culturing process morphological characterization was determined by SEM analyses. These scaffolds were designed to be nontoxic biomaterials. Here, a comparision was made between keratin containing 3T3, ECV and SAOS seeded nanofiber scaffolds and the results were presented and discussed.

Keywords: cell culture, keratin, nanofibers, UV-reactive electrospinning

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2852 Bamboo Resilience: Mentoring Asian Students to Develop their Self-Leadership via Online Seminars

Authors: Tam Nguyen

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Self-leadership is strongly tied to the ability to be resilient in the face of adversity. This study aims to demonstrate how a strategy based on a culturally relevant "bamboo metaphor" enables Asian students to cross cultural boundaries and to engage in online discussions to unlock their self-leadership potential. Asian students are influenced to varying degrees by the Confucian heritage culture, which educates students to respect authority, maintain harmony, and avoid public confrontations. This has a significant impact on the cultural readiness of Asian students to express their development as self-leaders. In this research project, researchers as mentors individually assist students, cultivate cognitive progress, encourage and personally ask students to join a process of mentorship program. This study analyzes and interprets the data from a large online seminar in Ho Chi Minh City, Vietnam, where students were trained in self-leadership skills. Focus-group interviews were implemented among 90 students in the program. Findings reveal the emotional needs of Asian students and suggest a cognitive model for developing students' self-awareness, self-confidence, and self-efficacy. The research results are anticipated to be applicable to a broader Asian population with a comparable cultural environment to Vietnam.

Keywords: self-leadership, bamboo resilience, cognitive modeling, Asian culture

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2851 Cross-Cultural Competence Development through 'Learning by Reflection': A Case Study of Chinese International Students Learning through Taking Part-Time Jobs in the UK

Authors: Xin Zhao

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The project aims to expand the notion of narrative learning and address the importance of learning by reflection in our learning and teaching context at a British university. Drawing on the key concepts such as development ZPD, transition and reflection-in and –on-action, this project analyses the learning experiences of a small sample of Chinese postgraduate students in a British University, who use part-time job experience to develop cross-cultural communication skills. The project adopts a mixed methods approach. Questionnaires and focus group interviews are used to examine the way in which students adapt (or not adapt) to the culture of learning in a British university and develop a renewed sense of self in transitions from one culture to the other. The project also looks at how the students appropriate opportunities for learning not just from classrooms but outside classrooms from everyday encounters. The project aims to address the implication of learning by reflection as development in transition. Time in and for learning, or duration, is taken for granted in theorising narrative learning. The project shall explore this very issue of time in relation to learning by reflection in considering time in/of/for learning as duration.

Keywords: cross-cultural competence, learning by refection, international student transition, part-time work experience

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2850 Hyaluronic Acid - Alginate Hydrogel for the Transdifferentiation of Testis Cells into Erythrocyte and Hepatocyte-like Cells; A Practice Within an Effective Agent Choice

Authors: Leila Rashki Ghaleno, Mohamad Amin Hajari, Leila Montazeri, Abdolhossein Shahverdi, Mojtaba Rezazadeh Valojerdi

Abstract:

Background: Spermatogonia stem cells (SSCs) exhibit pluripotency, enabling them to undergo differentiation into many cell lineages, including neurons, glia, endothelial cells, and hepatocytes when cultured in vitro. Although the specific mechanisms are not yet fully understood, it has been observed that biopolymer agents, such as hyaluronic acid (HA) and alginate (Alg), have the potential to induce transdifferentiation of SSCs. The current work aimed to examine the process of in vitro spermatogenesis and the conversion of mouse testicular cells into hepatocytes and erythrocyte-like cells utilizing the HA-Alg hydrogel. Method: After being extracted from the testes of a 5-day postpartum mouse (5 DPP), the testicular cells were separated into two enzymatic stages and then put into a composite hydrogel containing 0.5% HA and 1% alginate. On days 14 and 28 of culture, the colonies' growth, the cells' viability, and their histology were assessed. Result: Despite observing significant cell proliferation on day 14 and the development of circular-shaped organoids on day 28, it was noted that the organoids generated in the HA-Alg medium tended to maintain their circular morphology on day 28. Notably, the testicular cells underwent transdifferentiation into cell types resembling erythrocytes and hepatocytes. The hepatocyte-like cells exhibited the presence of glycogen and lipid deposits, indicating their hepatocyte-like characteristics. Interestingly, immunostaining analysis revealed the secretion of albumin and the presence of VEGFR on day 14. However, on day 28, albumin expression was not detected, while the expression of Sox9 (a marker for hepatocytes), Vegf, CD34, and C-kit (markers for erythrocytes) showed increased levels in the gene expression evaluation. Conclusion: The present findings indicated that HA-Alg could be a potent and effective agent for the transdifferentiation of testis cells into erythrocyte and hepatocyte-like cells, as recent studies have confirmed the transformation of SSCs into hepatocyte cells during in vitro culture.

Keywords: 3D culture, mouse testicular cell, hyaluronic acid, liver organoids

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2849 Safety Culture Implementation Based on Occupational Health and Safety Assessment

Authors: Nyambayar Davaadorj, Ichiro Koshijima

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Safety or the state of being safe can be described as a condition of being not dangerous or not harmful. It is necessary for an individual to avoid dangerous situations every day. Also, an organization is subject to legal requirements for the health and safety of persons inside and around the immediate workplace, or who are exposed to the workplace activities. Although it might be difficult to keep a situation where complete safety is ensured, efforts must nonetheless be made to consider ways of removing any potential danger within an organization. In order to ensure a safe working environment, the capability of responding (i.e., resilience) to signals (i.e., information concerning events that could pose future problems that must be taken into account) that occur in and around corporations is necessary. The ability to evaluate this essential point is thus one way in which safety and security can be managed. This study focuses on OHSAS18001, an internationally applied standard for the construction and operation of occupational health and safety management systems, by using IDEF0 for Function Modeling (IDEF0) and the Resilience Matrix originally made by Bracco. Further, this study discusses a method for evaluating a manner in which Occupational Health and Safety Assessment Series (OHSAS) systematically functions within corporations. Based on the findings, this study clarifies the potential structural objection for corporations when implementing and operating the OHSAS standard.

Keywords: OHSAS18001, IDEF0, resilience engineering, safety culture

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2848 The Convention of Culture: A Comprehensive Study on Dispute Resolution Pertaining to Heritage and Related Issues

Authors: Bhargavi G. Iyer, Ojaswi Bhagat

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In recent years, there has been a lot of discussion about ethnic imbalance and diversity in the international context. Arbitration is now subject to the hegemony of a small number of people who are constantly reappointed. When a court system becomes exclusionary, the quality of adjudication suffers significantly. In such a framework, there is a misalignment between adjudicators' preconceived views and the interests of the parties, resulting in a biased view of the proceedings. The world is currently witnessing a slew of intellectual property battles around cultural appropriation. The term "cultural appropriation" refers to the industrial west's theft of indigenous culture, usually for fashion, aesthetic, or dramatic purposes. Selena Gomez exemplifies cultural appropriation by commercially using the “bindi,” which is sacred to Hinduism, as a fashion symbol. In another case, Victoria's Secret insulted indigenous peoples' genocide by stealing native Indian headdresses. In the case of yoga, a similar process can be witnessed, with Vedic philosophy being reduced to a type of physical practice. Such a viewpoint is problematic since indigenous groups have worked hard for generations to ensure the survival of their culture, and its appropriation by the western world for purely aesthetic and theatrical purposes is upsetting to those who practise such cultures. Because such conflicts involve numerous jurisdictions, they must be resolved through international arbitration. However, these conflicts are already being litigated, and the aggrieved parties, namely developing nations, do not believe it prudent to use the World Intellectual Property Organization's (WIPO) already established arbitration procedure. This practise, it is suggested in this study, is the outcome of Europe's exclusionary arbitral system, which fails to recognise the non-legal and non-commercial nature of indigenous culture issues. This research paper proposes a more comprehensive, inclusive approach that recognises the non-legal and non-commercial aspects of IP disputes involving cultural appropriation, which can only be achieved through an ethnically balanced arbitration structure. This paper also aspires to expound upon the benefits of arbitration and other means of alternative dispute resolution (ADR) in the context of disputes pertaining to cultural issues; positing that inclusivity is a solution to the existing discord between international practices and localised cultural points of dispute. This paper also hopes to explicate measures that will facilitate ensuring inclusion and ideal practices in the domain of arbitration law, particularly pertaining to cultural heritage and indigenous expression.

Keywords: arbitration law, cultural appropriation, dispute resolution, heritage, intellectual property

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2847 Capacities of Early Childhood Education Professionals for the Prevention of Social Exclusion of Children

Authors: Dejana Bouillet, Vlatka Domović

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Both policymakers and researchers recognize that participating in early childhood education and care (ECEC) is useful for all children, especially for those who are exposed to the high risk of social exclusion. Social exclusion of children is understood as a multidimensional construct including economic, social, cultural, health, and other aspects of disadvantage and deprivation, which individually or combined can have an unfavorable effect on the current life and development of a child, as well as on the child’s development and on disadvantaged life chances in adult life. ECEC institutions should be able to promote educational approaches that portray developmental, cultural, language, and other diversity amongst children. However, little is known about the ways in which Croatian ECEC institutions recognize and respect the diversity of children and their families and how they respond to their educational needs. That is why this paper is dedicated to the analysis of the capacities of ECEC professionals to respond to the demands of educational needs of this very diverse group of children and their families. The results obtained in the frame of the project “Models of response to educational needs of children at risk of social exclusion in ECEC institutions,” funded by the Croatian Science Foundation, will be presented. The research methodology arises from explanations of educational processes and risks of social exclusion as a complex and heterogeneous phenomenon. The preliminary results of the qualitative data analysis of educational practices regarding capacities to identify and appropriately respond to the requirements of children at risk of social exclusion will be presented. The data have been collected by interviewing educational staff in 10 Croatian ECEC institutions (n = 10). The questions in the interviews were related to various aspects of inclusive institutional policy, culture, and practices. According to the analysis, it is possible to conclude that Croatian ECEC professionals are still faced with great challenges in the process of implementation of inclusive policies, culture, and practices. There are several baselines of this conclusion. The interviewed educational professionals are not familiar enough with the whole complexity and diversity of needs of children at risk of social exclusion, and the ECEC institutions do not have enough resources to provide all interventions that these children and their families need.

Keywords: children at risk of social exclusion, ECEC professionals, inclusive policies, culture and practices, quallitative analysis

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2846 Preparation and Application of Biocompatible Nanobioactive Glass as Therapeutic Agents for Bone Tissue Engineering

Authors: P. Shrivastava, S. Vijayalakshmi, A. K. Singh, S. Dalai, R. Teotia, P. Sharma, J. Bellare

Abstract:

This paper focuses on the synthesis and application of nanobioactive glass for bone regeneration studies. Nanobioactive glass has been synthesized by sol gel method having a combination of silicon, calcium and phosphorous in the molar ratio of 75:21:4. The prepared particles were analyzed for surface morphology by FEG SEM and FEG TEM. Physiochemical properties were investigated using ICP AES, FTIR spectroscopy and X-ray diffraction (XRD) techniques. To ascertain their use for therapeutic use, biocompatibility evaluation of the particles was done by performing soaking studies in SBF and in vitro cell culture studies on MG63 cell lines. Cell morphology was observed by FE SEM and phase contrast microscopy. Nanobioactive glasses (NBG) thus prepared were of 30-200 nm in size, which makes them suitable for nano-biomedical applications. The spherical shape of the particles imparts high surface to volume ratio, promoting fast growth of hydroxyapatite (HA), which is the mineral component of bone. As evaluated by in vitro cell culture studies the NBG was found to enhance the surface activation which enhances osteoblast adhesion. This is an essential parameter to improve bone tissue integration, thereby making nanobioactive glass therapeutically suitable for correcting bone defects.

Keywords: biocompatibility, bone tissue engineering, hydroxyapatite, nanobioactive glass

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2845 Irish Film Tourism, Neocolonialism and Star Wars: Charting a Course Towards Ecologically and Culturally Considered Representation and Tourism on Skellig Michael

Authors: Rachel Gough

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In 2014, Skellig Michael, an island off Ireland’s western seaboard and UNESCO world heritage site became a major setting in Disney’s Star Wars franchise. The subsequent influx of tourists to the site has proven to be a point of contention nationally. The increased visitor numbers have uplifted certain areas of the local economy, the mainland, but have caused irreparable damage to historic monuments and to endangered bird populations who breed on the island. Recent research carried out by a state body suggests far-reaching and longterm negative impacts on the island’s culture and environment, should the association with the Star Wars franchise persist. In spite of this, the film has been widely endorsed by the Irish government as providing a vital economic boost to historically marginalised rural areas through film tourism. This paper argues quite plainly that what is taking place on Skellig is neocolonialism. Skellig Michael’s unique resources, its aesthetic qualities, its ecosystem, and its cultural currency have been sold by the state to a multinational corporation, who profit from their use. Meanwhile, locals are left to do their best to turn a market trend into sustainable business at the expense of culture ecology and community. This paper intends to be the first dedicated study into the psychogeographic and cultural impact of Skellig Michael’s deterioration as a result of film tourism. It will discuss the projected impact of this incident on Irish culture more broadly and finally will attempt to lay out a roadmap for more collaborative filmmaking and touristic approach, which allows local cultures and ecosystem’s to thrive without drastically inhibiting cultural production. This paper will ultimately find that the consequences of this representation call for a requirement to read tourism as a split concept — namely into what we might loosely call “eco-tourism” and more capital-based “profit-bottom-line tourism.”

Keywords: ecology, film tourism, neocolonialism, sustainability

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2844 Efficiency of Pre-Treatment Methods for Biodiesel Production from Mixed Culture of Microalgae

Authors: Malith Premarathne, Shehan Bandara, Kaushalya G. Batawala, Thilini U. Ariyadasa

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The rapid depletion of fossil fuel supplies and the emission of carbon dioxide by their continued combustion have paved the way for increased production of carbon-neutral biodiesel from naturally occurring oil sources. The high biomass growth rate and lipid production of microalgae make it a viable source for biodiesel production compared to conventional feedstock. In Sri Lanka, the production of biodiesel by employing indigenous microalgae species is at its emerging stage. This work was an attempt to compare the various pre-treatment methods before extracting lipids such as autoclaving, microwaving and sonication. A mixed culture of microalgae predominantly consisting of Chlorella sp. was obtained from Beire Lake which is an algae rich, organically polluted water body located in Colombo, Sri Lanka. After each pre-treatment method, a standard solvent extraction using Bligh and Dyer’s method was used to compare the total lipid content in percentage dry weight (% dwt). The fatty acid profiles of the oils extracted with each pretreatment method were analyzed using gas chromatography-mass spectrometry (GC-MS). The properties of the biodiesels were predicted by Biodiesel Analyzer© Version 1.1, in order to compare with ASTM 6751-08 biodiesel standard.

Keywords: biodiesel, lipid extraction, microalgae, pre-treatment

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2843 Voices and Pictures from an Online Course and a Face to Face Course

Authors: Eti Gilad, Shosh Millet

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In light of the technological development and its introduction into the field of education, an online course was designed in parallel to the 'conventional' course for teaching the ''Qualitative Research Methods''. This course aimed to characterize learning-teaching processes in a 'Qualitative Research Methods' course studied in two different frameworks. Moreover its objective was to explore the difference between the culture of a physical learning environment and that of online learning. The research monitored four learner groups, a total of 72 students, for two years, two groups from the two course frameworks each year. The courses were obligatory for M.Ed. students at an academic college of education and were given by one female-lecturer. The research was conducted in the qualitative method as a case study in order to attain insights about occurrences in the actual contexts and sites in which they transpire. The research tools were open-ended questionnaire and reflections in the form of vignettes (meaningful short pictures) to all students as well as an interview with the lecturer. The tools facilitated not only triangulation but also collecting data consisting of voices and pictures of teaching and learning. The most prominent findings are: differences between the two courses in the change features of the learning environment culture for the acquisition of contents and qualitative research tools. They were manifested by teaching methods, illustration aids, lecturer's profile and students' profile.

Keywords: face to face course, online course, qualitative research, vignettes

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2842 Media Manipulations and the Culture of Beneficial Endophytic Fungi in the Leaves and Stem Bark of Grewia lasiocarpa E. Mey. Ex Harv

Authors: Akwu A. Nneka, Naidoo, Yougasphree

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A significantly high number of microbes exist in higher plants; these microbes include bacteria, fungi, and actinomycetes. There are reports on the benefits of endophytic fungi and their products of metabolism to the host plant and man, consequently, it is expedient to explore the changes that could arise as a result of manipulating their growth media. Grewia lasiocarpa E. Mey. ex Harv. (Malvaceae) is an indigenous Southern African plant, that belongs to a genus with known medicinal properties. Three media were used to culture the endophytic fungi viz., Potato Dextrose Agar (PDA), Malt Extract Agar (MEA), and Bacteriological Agar (BA) were used singly, and supplemented with three dilutions of the leaves and stem bark extracts. The manipulated growth media composition had a significant effect on the diversity of the isolated fungal populations. Several endophytic fungi were isolated; their distribution and diversity revealed a significant relatedness with the manipulated media. The media supplemented with the plant extracts was observed to give a significant increase in the growth rate and yield of the endophytes. To the best of our knowledge, this is the first study describing the endophytic fungi present in the leaves and stem bark of G. lasiocarpa E. Mey. ex Harv.

Keywords: Grewia lasiocarpa, plant-based extracts, endophytic fungi, Malvaceae

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2841 Study of the Architectural Heritage and Culture of Bene Israeli Community in Raigad, Maharashtra

Authors: Nikita Mahajani

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The Bene Israel is the oldest Jewish community on the subcontinent, and their shipwreck off the coast of Navgaon (a coastal settlement south of Alibaug), in which only seven men and women survived, anticipated the first millennia of their residence in the Konkan. The community was cut off from mainstream Judaism for decades as a result of its poverty. Almost all of the Bene Israel people in the northern Konkan region have relocated to Israel. The few who remained have settled down in Mumbai and Thane. Despite this, they have left a rich cultural legacy, as seen by their houses, cemeteries and synagogues. Even though the population has reduced owing to outside migration, much of this built legacy has survived. This paper attempts to examine the Jewish architectural heritage in Raigad with an aim to recreate their cultural, social and economic history. Oral histories by interviews of the Bene Israel community from Revdanda helped gain information about naming customs, migrations, professions, religious customs and funeral practices. The findings of this research reveal that most synagogues in Raigad district are shut due to a lack of Bene Israelis coming for prayers. The cemeteries are in a dilapidated condition. The little-known Bene Israeli community of Raigad is a seamless blend of Maharashtrian and Jew culture and feels more homely in India.

Keywords: Konkan, Alibag, Revdanda, Pen, Bene Israeli, Indian jews, synagogue, cemetry

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2840 Extracellular Production of the Oncolytic Enzyme, Glutaminase Free L-Asparaginase, from Newly Isolated Streptomyces Olivaceus NEAE-119: Optimization of Culture Conditions Using Response Surface Methodology

Authors: Noura El-Ahmady El-Naggar

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Among the antitumour drugs, bacterial enzyme L-asparaginase has been employed as the most effective chemotherapeutic agent in pediatric oncotherapy especially for acute lymphoblastic leukemia. Glutaminase free L-asparaginase producing actinomycetes were isolated from soil samples collected from Egypt. Among them, a potential culture, strain NEAE-119, was selected and identified on the basis of morphological, cultural, physiological and biochemical properties, together with 16S rDNA sequence as Streptomyces olivaceus NEAE-119 and sequencing product(1509 bp) was deposited in the GenBank database under accession number KJ200342. The optimization of different process parameters for L-asparaginase production by Streptomyces olivaceus NEAE-119 using Plackett–Burman experimental design and response surface methodology was carried out. Fifteen nutritional variables (temperature, pH, incubation time, inoculum size, inoculum age, agitation speed, dextrose, starch, L-asparagine, KNO3, yeast extract, K2HPO4, MgSO4.7H2O, NaCl and FeSO4. 7H2O) were screened using Plackett–Burman experimental design. The most positive significant independent variables affecting enzyme production (temperature, inoculum age and agitation speed) were further optimized by the central composite face-centered design -response surface methodology. As a result, a medium of the following formula is the optimum for producing an extracellular L-asparaginase in the culture filtrate of Streptomyces olivaceus NEAE-119: Dextrose 3g, starch 20g, L-asparagine 10g, KNO3 1g, K2HPO4 1g, MgSO4.7H2O 0.1g, NaCl 0.1g, pH 7, temperature 37°C, agitation speed 200 rpm/min, inoculum size 4%, v/v, inoculum age 72 h and fermentation period 5 days.

Keywords: Streptomyces olivaceus NEAE-119, glutaminase free L-asparaginase, production, Plackett-Burman design, central composite face-centered design, 16S rRNA, scanning electron microscope

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2839 Story Telling Method as a Bastion of Local Wisdom in the Frame of Education Technology Development in Medan, North Sumatra-Indonesia

Authors: Mardianto

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Education and learning are now grown rapidly. Synergy of techonology especially instructional technology in the learning activities are very big influence on the effectiveness of learning and creativity to achieve optimal results. But on the other hand there is a education value that is difficult to be articulated through character-forming technology such as honesty, discipline, hard work, heroism, and so forth. Learning strategy and storytelling from the past until today is still an option for teachers to convey the message of character values. With the material was loaded from the local culture (stories folklore), the combination of learning objectives (build character child) strategy, and traditional methods (storytelling and story), and the preservation of local culture (dig tale folklore) is critical to maintaining the nation's culture. In the context of maintaining the nation's culture, then since the age of the child at the level of government elementary school a necessity. Globalization, the internet and technology sometimes feel can displace the role of the teacher in the learning activities. To the oral tradition is a mainstay of storytelling should be maintained and preserved. This research was conducted at the elementary school in the city of Medan, North Sumatra Indonesia, with a random sampling technique, the 27 class teachers were respondents who were randomly assigned to the Madrasah Ibtdaiyah (Islamic Elementary School) both public and private. Research conducted at the beginning of 2014 refers to a curriculum that is being transformed in the environment ministry Republic Religion Indonesia. The results of this study indicate that; the declining skills of teachers to develop storytelling this can be seen from; 74.07% of teachers have never attended a special training storytelling, 85.19% no longer nasakah new stories, only 22.22% are teachers who incorporate methods of stories in the learning plan. Most teachers are no longer concerned with storytelling, among those experiencing difficulty in developing methods because the story; 66.67% of children are more interested in children's cartoons like Bobo boy, Angrybirds and others, 59.26 children prefer other activities than listening to a story. The teachers hope, folklore books should be preserved, storytelling training should be provided by the government through the ministry of religion, race or competition of storytelling should be scheduled, writing a new script-based populist storytelling should be provided immediately. The teachers’ hope certainly not excessive, by realizing the story method becomes articulation as the efforts of child character development based populist, therefore the local knowledge can be a strong fortress facing society in the era of progress as at present, and future.

Keywords: story telling, local wisdom, education, technology development

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2838 Comparison of Methods for the Detection of Biofilm Formation in Yeast and Lactic Acid Bacteria Species Isolated from Dairy Products

Authors: Goksen Arik, Mihriban Korukluoglu

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Lactic acid bacteria (LAB) and some yeast species are common microorganisms found in dairy products and most of them are responsible for the fermentation of foods. Such cultures are isolated and used as a starter culture in the food industry because of providing standardisation of the final product during the food processing. Choice of starter culture is the most important step for the production of fermented food. Isolated LAB and yeast cultures which have the ability to create a biofilm layer can be preferred as a starter in the food industry. The biofilm formation could be beneficial to extend the period of usage time of microorganisms as a starter. On the other hand, it is an undesirable property in pathogens, since biofilm structure allows a microorganism become more resistant to stress conditions such as antibiotic presence. It is thought that the resistance mechanism could be turned into an advantage by promoting the effective microorganisms which are used in the food industry as starter culture and also which have potential to stimulate the gastrointestinal system. Development of the biofilm layer is observed in some LAB and yeast strains. The resistance could make LAB and yeast strains dominant microflora in the human gastrointestinal system; thus, competition against pathogen microorganisms can be provided more easily. Based on this circumstance, in the study, 10 LAB and 10 yeast strains were isolated from various dairy products, such as cheese, yoghurt, kefir, and cream. Samples were obtained from farmer markets and bazaars in Bursa, Turkey. As a part of this research, all isolated strains were identified and their ability of biofilm formation was detected with two different methods and compared with each other. The first goal of this research was to determine whether isolates have the potential for biofilm production, and the second was to compare the validity of two different methods, which are known as “Tube method” and “96-well plate-based method”. This study may offer an insight into developing a point of view about biofilm formation and its beneficial properties in LAB and yeast cultures used as a starter in the food industry.

Keywords: biofilm, dairy products, lactic acid bacteria, yeast

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2837 Determinants of Corporate Social Responsibility Adoption: Evidence from China

Authors: Jing (Claire) LI

Abstract:

More than two decades from 2000 to 2020 of economic reforms have brought China unprecedented economic growth. There is an urgent call of research towards corporate social responsibility (CSR) in the context of China because while China continues to develop into a global trading market, it suffers from various serious problems relating to CSR. This study analyses the factors affecting the adoption of CSR practices by Chinese listed companies. The author proposes a new framework of factors of CSR adoption. Following common organisational factors and external factors in the literature (including organisational support, company size, shareholder pressures, and government support), this study introduces two additional factors, dynamic capability and regional culture. A survey questionnaire was conducted on the CSR adoption of Chinese listed companies in Shen Zhen and Shang Hai index from December 2019 to March 2020. The survey was conducted to collect data on the factors that affect the adoption of CSR. After collection of data, this study performed factor analysis to reduce the number of measurement items to several main factors. This procedure is to confirm the proposed framework and ensure the significant factors. Through analysis, this study identifies four grouped factors as determinants of the CSR adoption. The first factor loading includes dynamic capability and organisational support. The study finds that they are positively related to the first factor, so the first factor mainly reflects the capabilities of companies, which is one component in internal factors. In the second factor, measurement items of stakeholder pressures mainly are from regulatory bodies, customer and supplier, employees and community, and shareholders. In sum, they are positively related to the second factor and they reflect stakeholder pressures, which is one component of external factors. The third factor reflects organisational characteristics. Variables include company size and cultural score. Among these variables, company size is negatively related to the third factor. The resulted factor loading of the third factor implies that organisational factor is an important determinant of CSR adoption. Cultural consistency, the variable in the fourth factor, is positively related to the factor. It represents the difference between perception of managers and actual culture of the organisations in terms of cultural dimensions, which is one component in internal factors. It implies that regional culture is an important factor of CSR adoption. Overall, the results are consistent with previous literature. This study is of significance from both theoretical and empirical perspectives. First, from the significance of theoretical perspective, this research combines stakeholder theory, dynamic capability view of a firm, and neo-institutional theory in CSR research. Based on association of these three theories, this study introduces two new factors (dynamic capability and regional culture) to have a better framework for CSR adoption. Second, this study contributes to empirical literature of CSR in the context of China. Extant Chinese companies lack recognition of the importance of CSR practices adoption. This study built a framework and may help companies to design resource allocation strategies and evaluate future CSR and management practices in an early stage.

Keywords: China, corporate social responsibility, CSR adoption, dynamic capability, regional culture

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2836 Mindfulness in a Secular Age: Framing and Contextualising the Conversation in the Irish Context

Authors: Thomas P. Carroll

Abstract:

The phenomenon of mindfulness has become ever more popular in an increasingly pluralist Western society. Mindfulness practice has penetrated secular contexts that would otherwise be closed to religious influence, including state schools, hospitals, and commerce. The contemporary understanding of mindfulness has its origins in Buddhist meditation. However, since Jon Kabat-Zinn’s pioneering work in Mindfulness-Based Interventions, the concept has developed and sometimes mutated into various forms of practice which are disembedded from their original spiritual philosophy. This project will explore the spiritual climate within which mindfulness is currently flourishing through dialogue with three interlocutors. The first interlocutor is the Canadian philosopher Charles Taylor whose seminal work, ‘A Secular Age’, outlines three distinct modes of secularity. Taylor examines how the conditions of belief have changed and how the self seeks meaning in an age where belief in the divine is no longer axiomatic. The next interlocutor is Czech theologian and psychotherapist Tomáš Halík who offers a unique perspective of a Catholic who belongs to a section of society outnumbered by secular counterparts, with a theological hermeneutic best described as 'Den Fremden verstehen- understanding the stranger'. Finally, Irish theologian Michael Paul Gallagher offers a theological perspective on how the Christian faith can be translated into dialogue with Irish secular culture, as well as addressing the crisis of imagination and culture rather than the crisis of faith in Ireland. These interlocutors will illustrate that there are sometimes striking differences in how to interpret the religious signs of the times. However, these approaches also reveal significant similarities in how they address and explore the meaning of religious belief and experience today. In this way, themes will emerge that will help to frame the conversation about mindfulness in the West. These themes will include; the failure of the secularization thesis to pass, the growth of a diverse marketplace of religions and beliefs and the growth of a demographic who identify as spiritual but not religious. Such research is paramount in enabling a richer dialogue between Christian faith and mindfulness in a fragmented, postmodern Western context.

Keywords: culture, mindfulness, secularism, spirituality

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2835 Microfluidic Fluid Shear Mechanotransduction Device Using Linear Optimization of Hydraulic Channels

Authors: Sanat K. Dash, Rama S. Verma, Sarit K. Das

Abstract:

A logarithmic microfluidic shear device was designed and fabricated for cellular mechanotransduction studies. The device contains four cell culture chambers in which flow was modulated to achieve a logarithmic increment. Resistance values were optimized to make the device compact. The network of resistances was developed according to a unique combination of series and parallel resistances as found via optimization. Simulation results done in Ansys 16.1 matched the analytical calculations and showed the shear stress distribution at different inlet flow rates. Fabrication of the device was carried out using conventional photolithography and PDMS soft lithography. Flow profile was validated taking DI water as working fluid and measuring the volume collected at all four outlets. Volumes collected at the outlets were in accordance with the simulation results at inlet flow rates ranging from 1 ml/min to 0.1 ml/min. The device can exert fluid shear stresses ranging four orders of magnitude on the culture chamber walls which will cover shear stress values from interstitial flow to blood flow. This will allow studying cell behavior in the long physiological range of shear stress in a single run reducing number of experiments.

Keywords: microfluidics, mechanotransduction, fluid shear stress, physiological shear

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2834 A Genetic Identification of Candida Species Causing Intravenous Catheter-Associated Candidemia in Heart Failure Patients

Authors: Seyed Reza Aghili, Tahereh Shokohi, Shirin Sadat Hashemi Fesharaki, Mohammad Ali Boroumand, Bahar Salmanian

Abstract:

Introduction: Intravenous catheter-associated fungal infection as nosocomial infection continue to be a deep problem among hospitalized patients, decreasing quality of life and adding healthcare costs. The capacity of catheters in the spread of candidemia in heart failure patients is obvious. The aim of this study was to evaluate the prevalence and genetic identification of Candida species in heart disorder patients. Material and Methods: This study was conducted in Tehran Hospital of Cardiology Center (Tehran, Iran, 2014) during 1.5 years on the patients hospitalized for at least 7 days and who had central or peripheral vein catheter. Culture of catheters, blood and skin of the location of catheter insertion were applied for detecting Candida colonies in 223 patients. Identification of Candida species was made on the basis of a combination of various phenotypic methods and confirmed by sequencing the ITS1-5.8S-ITS2 region amplified from the genomic DNA using PCR and the NCBI BLAST. Results: Of the 223 patients samples tested, we identified totally 15 Candida isolates obtained from 9 (4.04%) catheter cultures, 3 (1.35%) blood cultures and 2 (0.90%) skin cultures of the catheter insertion areas. On the base of ITS region sequencing, out of nine Candida isolates from catheter, 5(55.6%) C. albicans, 2(22.2%) C. glabrata, 1(11.1%) C. membranifiaciens and 1 (11.1%) C. tropicalis were identified. Among three Candida isolates from blood culture, C. tropicalis, C. carpophila and C. membranifiaciens were identified. Non-candida yeast isolated from one blood culture was Cryptococcus albidus. One case of C. glabrata and one case of Candida albicans were isolated from skin culture of the catheter insertion areas in patients with positive catheter culture. In these patients, ITS region of rDNA sequence showed a similarity between Candida isolated from the skin and catheter. However, the blood samples of these patients were negative for fungal growth. We report two cases of catheter-related candidemia caused by C. membranifiaciens and C. tropicalis on the base of genetic similarity of species isolated from blood and catheter which were treated successfully with intravenous fluconazole and catheter removal. In phenotypic identification methods, we could only identify C. albicans and C. tropicalis and other yeast isolates were diagnosed as Candida sp. Discussion: Although more than 200 species of Candida have been identified, only a few cause diseases in humans. There is some evidence that non-albicans infections are increasing. Many risk factors, including prior antibiotic therapy, use of a central venous catheter, surgery, and parenteral nutrition are considered to be associated with candidemia in hospitalized heart failure patients. Identifying the route of infection in candidemia is difficult. Non-albicans candida as the cause of candidemia is increasing dramatically. By using conventional method, many non-albicans isolates remain unidentified. So, using more sensitive and specific molecular genetic sequencing to clarify the aspects of epidemiology of the unknown candida species infections is essential. The positive blood and catheter cultures for candida isolates and high percentage of similarity of their ITS region of rDNA sequence in these two patients confirmed the diagnosis of intravenous catheter-associated candidemia.

Keywords: catheter-associated infections, heart failure patient, molecular genetic sequencing, ITS region of rDNA, Candidemia

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2833 International Criminal Prosecution and Core International Crimes

Authors: Ikediobi Lottanna Samuel

Abstract:

Days are gone when perpetrators of core international crimes hide under the cloak of sovereignty to go with impunity. The principle of international criminal responsibility is a reality. This move to end impunity for violation of human rights has led to the creation of international and hybrid tribunals, a permanent international criminal court, and increased prosecution of human rights violations in domestic courts. This article examines the attempts by the international community to bring perpetrators of heinous crimes to book. The work reveals the inadequacy of the current international mechanism for prosecuting core international crimes in order to end the culture of impunity and entrench the culture of accountability. It also identifies that ad hoc international criminal tribunals and the international criminal court face similar challenges ranging from lack of cooperation by nation states, non-existence of hierarchy of crimes, lack of effective enforcement mechanism, limited prosecutorial capacity and agenda, difficulty in apprehending suspects, difficulty in blending different legal tradition, absence of a coherent sentencing guideline, distant location of courts, selective indictment, etc. These challenges adversely affect the functioning of these courts. It is suggested that a more helpful way to end impunity would be to have a more robust and synergistic relationship between national, regional, and international approaches to prosecuting core international crimes.

Keywords: prosecution, criminal, international, tribunal, justice, ad hoc

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2832 The Integration and Practice of Indigenous Knowledge System and Sustainable Environmental Education Concept

Authors: Shih-Tsung Chen, Yenchin Hsiao

Abstract:

Evergreen Lily is a newly-built school after Morakot Typhoon took place. The school is located on Majia farm, which is surrounded by mountains. The fund in the construction of the school is solely sponsored by Chang Yung-Fa Foundation. There are 483 permanent houses near the school belonging to three tribes, Dashe, Majia, and Haocha. Due to the most ancient heritages of Paiwan and Rukai in these three tribes, the school is full of cultural atmosphere. From modern and traditional perspectives, Evergreen Lily strives to establish and develop a long-lasting educational model to meet the expectation of the tribes, parents, and the public. This study is a case study of how to develop indigenous education in newly established schools after the Morakot Hurricane disaster to meet the concept of environmental education. The systematic curriculum construction of education and cultural integration and the systematic practice of curriculum practice will be discussed, and the concept and practice of tribal education curriculum and sustainable environmental education will be understood. This study found that this school integrates the spirit of natural philosophy, democratic education, ethnic and experimental education, and constructs a knowledge system that includes three levels of spiritual culture, institutional culture, and material culture, as well as six dimensions of life philosophy, natural ecology, organizational system, tribal literature and history, song and dance, and technical and artistic methods. Adhering to the concept of harmonious education and the sustainable common good, the development of school-based tribal academic courses accounts for about one-third of the total number of teaching sessions, and there are different cultural themes in grades one to six, and there are clear teaching modules to effectively enhance students' potential inspiration. The complete curriculum implementation model can be described as a model for the development of indigenous schools to sustainable environmental education.

Keywords: environmental education, indigenous education, sustainable development, school-based curriculum

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2831 Evaluation of Collagen Synthesis in Macrophages/Fibroblasts Co-Culture Using Polylactic Acid Particles as Stimulants

Authors: Feng Ju Chuang, Yu Wen Wang, Tai Jung Hsieh, Shyh Ming Kuo

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Polylactic acid is a synthetic polymer with good biocompatibility and degradability, is widely used in clinical applications. In this study, we utilized Polylactic acid particles as stimulants for macrophages and the collagen synthesis of co-cultured fibroblasts was evaluated. The results indicated that Polylactic acid particles were nontoxic to cells from 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide. No obvious inflammation effect was observed (under the PLLA concentration of 1 mg/mL) after 24-h co-culture of Raw264.7 and NIH3T3 cells (from TNF-α assay). The addition of PLLA particles to the Raw264.7 and NIH3T3 co-cultures increased the synthesis of collagen, the highest collagen synthesis from the fibroblast was the 0.2 mg/mL (approximately 60% increased as compared with without addition Polylactic acid particles). Moreover, a co-axial atomization delivery device was used to percutaneously introduce Polylactic acid particles into the dermis layer and stimulating macrophages to secrete growth factors promoting fibroblasts to produce collagen. The preliminary results demonstrated the synthesis of collagen was increased mildly after the introduction of Polylactic acid particles for 28-d post implantation. The Polylactic acid particles could be successfully introduced into the dermis layer from H&E staining examination, however, the optimum concentration of Polylactic acid particles and the time-period for collagen synthesis still need to be evaluated.

Keywords: collagen synthesis, macrophage, NIH3T3 cells, polylactic acid particles

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2830 The Impact of Artificial Intelligence on Rural Life

Authors: Triza Edwar Fawzi Deif

Abstract:

In the process of urbanization in China, new rural construction is on the ascendant, which is becoming more and more popular. Under the driving effect of rural urbanization, the house pattern and tectonic methods of traditional vernacular houses have shown great differences from the family structure and values of contemporary peasant families. Therefore, it is particularly important to find a prototype, form and strategy to make a balance between the traditional memory and modern functional requirements. In order for research to combine the regional culture with modern life, under the situation of the current batch production of new rural residences, Badie village, in Zhejiang province, is taken as the case. This paper aims to put forward a prototype which can not only meet the demand of modern life but also ensure the continuation of traditional culture and historical context for the new rural dwellings design. This research not only helps to extend the local context in the construction of the new site but also contributes to the fusion of old and new rural dwellings in the old site construction. Through the study and research of this case, the research methodology and results can be drawn as reference for the new rural construction in other areas.

Keywords: steel slag, co-product, primary coating, steel aggregate capital, rural areas, rural planning, rural governance village, design strategy, new rural dwellings, regional context, regional expression

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2829 Experimental Investigation of the Impact of Biosurfactants on Residual-Oil Recovery

Authors: S. V. Ukwungwu, A. J. Abbas, G. G. Nasr

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The increasing high price of natural gas and oil with attendant increase in energy demand on world markets in recent years has stimulated interest in recovering residual oil saturation across the globe. In order to meet the energy security, efforts have been made in developing new technologies of enhancing the recovery of oil and gas, utilizing techniques like CO2 flooding, water injection, hydraulic fracturing, surfactant flooding etc. Surfactant flooding however optimizes production but poses risk to the environment due to their toxic nature. Amongst proven records that have utilized other type of bacterial in producing biosurfactants for enhancing oil recovery, this research uses a technique to combine biosurfactants that will achieve a scale of EOR through lowering interfacial tension/contact angle. In this study, three biosurfactants were produced from three Bacillus species from freeze dried cultures using sucrose 3 % (w/v) as their carbon source. Two of these produced biosurfactants were screened with the TEMCO Pendant Drop Image Analysis for reduction in IFT and contact angle. Interfacial tension was greatly reduced from 56.95 mN.m-1 to 1.41 mN.m-1 when biosurfactants in cell-free culture (Bacillus licheniformis) were used compared to 4. 83mN.m-1 cell-free culture of Bacillus subtilis. As a result, cell-free culture of (Bacillus licheniformis) changes the wettability of the biosurfactant treatment for contact angle measurement to more water-wet as the angle decreased from 130.75o to 65.17o. The influence of microbial treatment on crushed rock samples was also observed by qualitative wettability experiments. Treated samples with biosurfactants remained in the aqueous phase, indicating a water-wet system. These results could prove that biosurfactants can effectively change the chemistry of the wetting conditions against diverse surfaces, providing a desirable condition for efficient oil transport in this way serving as a mechanism for EOR. The environmental friendly effect of biosurfactants applications for industrial purposes play important advantages over chemically synthesized surfactants, with various possible structures, low toxicity, eco-friendly and biodegradability.

Keywords: bacillus, biosurfactant, enhanced oil recovery, residual oil, wettability

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2828 Examining the European Central Bank's Marginal Attention to Human Rights Concerns during the Eurozone Crisis through the Lens of Organizational Culture

Authors: Hila Levi

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Respect for human rights is a fundamental element of the European Union's (EU) identity and law. Surprisingly, however, the protection of human rights has been significantly restricted in the austerity programs ordered by the International Monetary Fund (IMF), the European Central Bank (ECB) and the European Commission (EC) (often labeled 'the Troika') in return for financial aid to the crisis-hit countries. This paper focuses on the role of the ECB in the crisis management. While other international financial institutions, such as the IMF or the World Bank, may opt to neglect human rights obligations, one might expect a greater respect of human rights from the ECB, which is bound by the EU Charter of Fundamental Rights. However, this paper argues that ECB officials made no significant effort to protect human rights or strike an adequate balance between competing financial and human rights needs while coping with the crisis. ECB officials were preoccupied with the need to stabilize the economy and prevent a collapse of the Eurozone, and paid only marginal attention to human rights concerns in the design and implementation of Troikas' programs. This paper explores the role of Organizational Culture (OC) in explaining this marginalization. While International Relations (IR) research on Intergovernmental Organizations (IGOs) behavior has traditionally focused on external interests of powerful member states, and on national and economic considerations, this study focuses on particular institutions' internal factors and independent processes. OC characteristics have been identified in OC literature as an important determinant of organizational behavior. This paper suggests that cultural characteristics are also vital for the examination of IGOs, and particularly for understanding the ECB's behavior during the crisis. In order to assess the OC of the ECB and the impact it had on its policies and decisions during the Eurozone crisis, the paper uses the results of numerous qualitative interviews conducted with high-ranking officials and staff members of the ECB involved in the crisis management. It further reviews primary sources of the ECB (such as ECB statutes, and the Memoranda of Understanding signed between the crisis countries and the Troika), and secondary sources (such as the report of the UN High Commissioner for Human Rights on Austerity measures and economic, social, and cultural rights). It thus analyzes the interaction between the ECBs culture and the almost complete absence of human rights considerations in the Eurozone crisis resolution scheme. This paper highlights the importance and influence of internal ideational factors on IGOs behavior. From a more practical perspective, this paper may contribute to understanding one of the obstacles in the process of human rights implementation in international organizations, and provide instruments for better protection of social and economic rights.

Keywords: European central bank, eurozone crisis, intergovernmental organizations, organizational culture

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2827 Deciphering Tumor Stroma Interactions in Retinoblastoma

Authors: Rajeswari Raguraman, Sowmya Parameswaran, Krishnakumar Subramanian, Jagat Kanwar, Rupinder Kanwar

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Background: Tumor microenvironment has been implicated in several cancers to regulate cell growth, invasion and metastasis culminating in outcome of therapy. Tumor stroma consists of multiple cell types that are in constant cross-talk with the tumor cells to favour a pro-tumorigenic environment. Not much is known about the existence of tumor microenvironment in the pediatric intraocular malignancy, Retinoblastoma (RB). In the present study, we aim to understand the multiple stromal cellular subtypes and tumor stromal interactions expressed in RB tumors. Materials and Methods: Immunohistochemistry for stromal cell markers CD31, CD68, alpha-smooth muscle (α-SMA), vimentin and glial fibrillary acidic protein (GFAP) was performed on formalin fixed paraffin embedded tissues sections of RB (n=12). The differential expression of stromal target molecules; fibroblast activation protein (FAP), tenascin-C (TNC), osteopontin (SPP1), bone marrow stromal antigen 2 (BST2), stromal derived factor 2 and 4 (SDF2 and SDF4) in primary RB tumors (n=20) and normal retina (n=5) was studied by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) and Western blotting. The differential expression was correlated with the histopathological features of RB. The interaction between RB cell lines (Weri-Rb-1, NCC-RbC-51) and Bone marrow stromal cells (BMSC) was also studied using direct co-culture and indirect co-culture methods. The functional effect of the co-culture methods on the RB cells was evaluated by invasion and proliferation assays. Global gene expression was studied by using Affymetrix 3’ IVT microarray. Pathway prediction was performed using KEGG and the key molecules were validated using qRT-PCR. Results: The immunohistochemistry revealed the presence of several stromal cell types such as endothelial cells (CD31+;Vim+/-); macrophages (CD68+;Vim+/-); Fibroblasts (Vim+; CD31-;CD68- );myofibroblasts (α-SMA+/ Vim+) and invading retinal astrocytes/ differentiated retinal glia (GFAP+; Vim+). A characteristic distribution of these stromal cell types was observed in the tumor microenvironment, with endothelial cells predominantly seen in blood vessels and macrophages near actively proliferating tumor or necrotic areas. Retinal astrocytes and glia were predominant near the optic nerve regions in invasive tumors with sparse distribution in tumor foci. Fibroblasts were widely distributed with rare evidence of myofibroblasts in the tumor. Both gene and protein expression revealed statistically significant (P<0.05) up-regulation of FAP, TNC and BST2 in primary RB tumors compared to the normal retina. Co-culture of BMSC with RB cells promoted invasion and proliferation of RB cells in direct and indirect contact methods respectively. Direct co-culture of RB cell lines with BMSC resulted in gene expression changes in ECM-receptor interaction, focal adhesion, IL-8 and TGF-β signaling pathways associated with cancer. In contrast, various metabolic pathways such a glucose, fructose and amino acid metabolism were significantly altered under the indirect co-culture condition. Conclusion: The study suggests that the close interaction between RB cells and the stroma might be involved in RB tumor invasion and progression which is likely to be mediated by ECM-receptor interactions and secretory factors. Targeting the tumor stroma would be an attractive option for redesigning treatment strategies for RB.

Keywords: gene expression profiles, retinoblastoma, stromal cells, tumor microenvironment

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