Search results for: illuminated both culture

Authors: Jihong Liang, Huiling Feng, Linqing Ma, Tianjiao Qi

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Confucius is the first sage in Chinese culture. Confucianism, the theories represented by Confucius, has long been at the core of Chinese traditional society, as the dominating political ideology of centralized feudal monarchy for more than two thousand years. Confucius Worship Ceremony held in the Confucian Temple in Qufu (Confucius’s birthplace), which is dedicated to commemorate Confucius and other 170 elites in Confucianism with a whole set of formal rites, pertains to “Auspicious Rites”, which worship heaven and earth, humans and ghosts. It was first a medium-scaled ritual activity but then upgraded to the supreme one at national level in the Qing Dynasty. As a national event, it was celebrated by Emperor as well as common intellectuals in traditional China. The Ceremony can be solemn and respectful, with prescribed and complicated procedures, well-prepared utensil and matched offerings operated in rhythm with music and dances. Each participant has his place, and everyone follows the specified rules. This magnificent ritual Ceremony, while embedded with rich culture connotation, actually symbolizes the social acknowledgment for orthodox culture represented by Confucianism. Rites reflected in this Ceremony, is one of the most important features of Chinese culture, serving as the key bond in the identification and continuation of Chinese culture. These rites and ritual ceremonies, as culture memories themselves, are not only treasures of China, but of the whole world. However, while the ancient Chinese Rite has been one of the thorniest and most complicated topics for academics, the more regrettable is that due to their interruption in practice and historical changes, these rites and ritual ceremonies have already become a vague language in today’s academic discourse and strange terms of the past for common people. Luckily, we, today, by virtue of modern digital technology, may be able to reproduce these ritual ceremonies, as most of them can still be found in ancient manuscripts, through which Chinese ancestors tell the beauty and gravity of their dignified rites and more importantly, their spiritual pursuits with vivid language and lively pictures. This research, based on review and interpretation of the ancient literature, intends to construct the ancient ritual ceremonies, with the Confucius Worship Ceremony as a case and by use of digital technology. Using 3D technology, the spatial scenes in the Confucian Temple can be reconstructed by virtual reality; the memorial tablet exhibited in the temple by GIS and different rites in the ceremonies by animation technology. With reference to the lyrics, melodies and lively pictures recorded in ancient scripts, it is also possible to reproduce the live dancing site. Also, image rendering technology can help to show the life experience and accomplishments of Confucius. Finally, lining up all the elements in a multimedia narrative form, a complete digitalized Confucius Worship Ceremony can be reproduced, which will provide an excellent virtual experience that goes beyond time and space by bringing its audience back to that specific historical time. This digital project, once completed, will play an important role in the inheritance and dissemination of cultural heritage.

Keywords: Confucius worship ceremony, multimedia narrative form, GIS, visual representation

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Authors: Tahisa M. Pedroso, Hérida R. N. Salgado

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The microbiological turbidimetric assay allows the determination of potency of the drug, by measuring the turbidity (absorbance), caused by inhibition of microorganisms by ertapenem sodium. Ertapenem sodium (ERTM), a synthetic antimicrobial agent of the class of carbapenems, shows action against Gram-negative, Gram-positive, aerobic and anaerobic microorganisms. Turbidimetric assays are described in the literature for some antibiotics, but this method is not described for ertapenem. The objective of the present study was to develop and validate a simple, sensitive, precise and accurate microbiological assay by turbidimetry to quantify ertapenem sodium injectable as an alternative to the physicochemical methods described in the literature. Several preliminary tests were performed to choose the following parameters: Staphylococcus aureus ATCC 25923, IAL 1851, 8 % of inoculum, BHI culture medium, and aqueous solution of ertapenem sodium. 10.0 mL of sterile BHI culture medium were distributed in 20 tubes. 0.2 mL of solutions (standard and test), were added in tube, respectively S1, S2 and S3, and T1, T2 and T3, 0.8 mL of culture medium inoculated were transferred to each tube, according parallel lines 3 x 3 test. The tubes were incubated in shaker Marconi MA 420 at a temperature of 35.0 °C ± 2.0 °C for 4 hours. After this period, the growth of microorganisms was inhibited by addition of 0.5 mL of 12% formaldehyde solution in each tube. The absorbance was determined in Quimis Q-798DRM spectrophotometer at a wavelength of 530 nm. An analytical curve was constructed to obtain the equation of the line by the least-squares method and the linearity and parallelism was detected by ANOVA. The specificity of the method was proven by comparing the response obtained for the standard and the finished product. The precision was checked by testing the determination of ertapenem sodium in three days. The accuracy was determined by recovery test. The robustness was determined by comparing the results obtained by varying wavelength, brand of culture medium and volume of culture medium in the tubes. Statistical analysis showed that there is no deviation from linearity in the analytical curves of standard and test samples. The correlation coefficients were 0.9996 and 0.9998 for the standard and test samples, respectively. The specificity was confirmed by comparing the absorbance of the reference substance and test samples. The values obtained for intraday, interday and between analyst precision were 1.25%; 0.26%, 0.15% respectively. The amount of ertapenem sodium present in the samples analyzed, 99.87%, is consistent. The accuracy was proven by the recovery test, with value of 98.20%. The parameters varied did not affect the analysis of ertapenem sodium, confirming the robustness of this method. The turbidimetric assay is more versatile, faster and easier to apply than agar diffusion assay. The method is simple, rapid and accurate and can be used in routine analysis of quality control of formulations containing ertapenem sodium.

Keywords: ertapenem sodium, turbidimetric assay, quality control, validation

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Authors: Dane Morace-Court

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This paper offers a reflective and reflexive examination of the lived experience of a group of young, white, working-class males engaging in secondary-education in England at a time when this population is widely recognised as the lowest attaining ethnic group within British schools. The focus of the paper is an exploration of the development of identities and aspirations, alongside contemporary demographic and ideological shifts in the British population, in their intersection with neoliberal education policies and the emerging ideological conflict between identity conservatism and liberalism. The construction and performance of intersecting social-class, gender, ethnic and national identities is considered as well as the process through which socially constructed narratives inform identities, values, and aspirations. Evocative autoethnography is then employed to offer reflections on working-class habitus and, in particular, classed and gendered codes that underpin expectations of manhood in post-industrial culture within an education system which seemingly requires the abandonment of aspects of a working-class background. Findings from the study identify the emergence of a culture of hyper-individualisation amongst white, working-class males in schools and a belief in the meritocratic ideologies of the New Right. In particular, the breakdown of the social contract, including notions of political and civic responsibility, coupled with the symbolic violence perpetrated against working-class culture and solidarity in British schools, have all informed the construction of a working-class masculinity which values the individual entrepreneur over the collective, and depoliticizes students to an extent where a focus on the spectacle and performance of success has replaced individual and collective investment in community.

Keywords: education, identity, masculinity, neoliberalism, working-class, intersectionality, autoethnography

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Authors: Nazirah Hassan, Andrew Kendrick

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Punishment inside the penal institution has always been practiced in order to maintain discipline and keep order. Nonetheless, criminologists have long debated that the enforcement of discipline by punishing inmates is often ineffective and has a detrimental impact on inmates’ conduct. This paper uses data from a sample of 289 incarcerated young offenders to investigate the prevalence of institutional misconduct. It explores punitive cultural practices inside institutions and how this culture affects the inmates’ conduct during confinement. The project focused on male and female young offenders aged 12 to 21 years old, in eight juvenile justice institutions. The research collected quantitative and qualitative data using a mixed-method approach. All participants completed the Direct and Indirect Prisoner behavior Checklist-Scaled Version Revised (DIPC-SCALED-R). In addition, exploratory interviews were carried out with sixteen inmates and eight institutional staff. Results of the questionnaire survey show that almost half of the inmates reported a higher level of involvement in perpetration. It demonstrates a remarkable convergence of direct, rather than indirect, perpetration. Also, inmates reported a higher level of tobacco used and behavior associated with negative attitudes towards staff and institutional rules. In addition to this, the qualitative data suggests that the punitive culture encourages the onset of misconduct by increasing the stressful and oppressive conditions within the institution. In general, physical exercise and locking up inmates are two forms of punishment that were ubiquitous throughout the institutions. Interestingly, physical exercise is not only enforced by institutional staff but also inmates. These findings are discussed in terms of existing literature and their practical implications are considered.

Keywords: institutional punishment, incarcerated young offenders, punitive culture, institutional misconduct

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Authors: Ainur Mukhambetova, Miras Karzhauov, Vyacheslav Ogay

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Introduction: Mesenchymal stem cells (MSCs) have the capacity to be differentiated into several cell lineages and are a promising source for cell therapy and tissue engineering. However, currently most MSCs culturing protocols use media supplemented with fetal bovine serum (FBS), which limits their application in clinic due to the possibility of zoonotic infections, contamination and immunological reactions. Consequently, formulating effective serum free culture medium becomes one of the important problems in contemporary cell biotechnology. Objectives: The aim of this study was to define an optimal serum-free medium for culturing of periosteum derived MSCs. Materials and methods: The MSCs were extracted from human periosteum and transferred to the culture flasks pretreated with CELLstart™. Immunophenotypic characterization, proliferation and in vitro differentiation of cells grown on STEM PRO® MSC SFM were compared to the cells cultured in the standard FBS containing media. Chromosome analysis and flow cytometry were also performed. Results: We have shown that cells were grown on STEM PRO® MSC SFM retained all the morphological, immunophenotypic (CD73, CD90, CD105, vimentin and Stro-1) and cell differentiation characteristics specific to MSCs. Chromosome analysis indicated no anomalies in the chromosome structure. Flow cytometry showed a high expression of cell adhesion molecules CD44 (98,8%), CD90 (97,4%), CD105 (99,1%). In addition, we have shown that cell is grown on STEM PRO® MSC SFM have higher proliferation capacity compared to cell expanded on standard FBS containing the medium. Conclusion: We have shown that STEM PRO® MSC SFM is optimal for culturing periosteum derived human MSCs which subsequently can be safely used in cell therapy.

Keywords: cell technologies, periosteum-derived MSCs, regenerative medicine, serum-free medium

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Authors: Qingyu Zhang, Mei Cao

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As the environments become increasingly uncertain, firms have attempted to achieve greater supply chain collaboration. Supply chain collaboration can generate significant benefits to its members, e.g., reducing risks and decreasing transaction costs. However, a strong relationship is often related to firm’s culture (e.g., short-term vs. long-term interests). The objective of the study is to explore the effect of long-term oriented culture on product quality in a supply chain. Data was collected through a Web survey of U.S. manufacturing firms. Structural equation modeling (LISREL) was used to analyze the data. The results support the mediating roles of goal congruence and communication in the relationship between long-term orientation and product quality in the supply chain. Goal congruence partially mediates the relationship between long-term orientation and communication; communication completely mediates the relationship between goal congruence and product quality. Without high levels of communication, goal congruence cannot improve product quality in a positive way.

Keywords: communication, long-term orientation, product quality, supply chain

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Authors: Urvi Panwar, Kanchan Mishra, Kanjaksha Ghosh, ShankerLal Kothari

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Background: Day-by-day the increasing prevalence of neurodegenerative diseases have become a global issue to manage them by medical sciences. The adult neural stem cells are rare and require an invasive and painful procedure to obtain it from central nervous system. Mesenchymal stem cell (MSCs) therapies have shown remarkable application in treatment of various cell injuries and cell loss. MSCs can be derived from various sources like adult tissues, human bone marrow, umbilical cord blood and cord tissue. MSCs have similar proliferation and differentiation capability, but the human umbilical cord-derived mesenchymal stem cells (hUCMSCs) are proved to be more beneficial with respect to cell procurement, differentiation to other cells, preservation, and transplantation. Material and method: Human umbilical cord is easily obtainable and non-controversial comparative to bone marrow and other adult tissues. The umbilical cord can be collected after delivery of baby, and its tissue can be cultured using explant culture method. Cell culture medium such as DMEMF12+10% FBS and DMEMF12+Neural growth factors (bFGF, human noggin, B27) with antibiotics (Streptomycin/Gentamycin) were used to culture and differentiate mesenchymal stem cells into neural cells, respectively. The characterisations of MSCs were done with Flow Cytometer for surface markers CD90, CD73 and CD105 and colony forming unit assay. The differentiated various neural cells will be characterised by fluorescence markers for neurons, astrocytes, and oligodendrocytes; quantitative PCR for genes Nestin and NeuroD1 and Western blotting technique for gap43 protein. Result and discussion: The high quality and number of MSCs were isolated from human umbilical cord via explant culture method. The obtained MSCs were differentiated into neural cells like neurons, astrocytes and oligodendrocytes. The differentiated neural cells can be used to treat neural injuries and neural cell loss by delivering cells by non-invasive administration via cerebrospinal fluid (CSF) or blood. Moreover, the MSCs can also be directly delivered to different injured sites where they differentiate into neural cells. Therefore, human umbilical cord is demonstrated to be an inexpensive and easily available source for MSCs. Moreover, the hUCMSCs can be a potential source for neural cell therapies and neural cell regeneration for neural cell injuries and neural cell loss. This new way of research will be helpful to treat and manage neural cell damages and neurodegenerative diseases like Alzheimer and Parkinson. Still the study has a long way to go but it is a promising approach for many neural disorders for which at present no satisfactory management is available.

Keywords: bone marrow, cell therapy, explant culture method, flow cytometer, human umbilical cord, mesenchymal stem cells, neurodegenerative diseases, neuroprotective, regeneration

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Authors: Dieter Veldsman, Theo Heyns Veldsman

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The knowledge economy demands an organisation that is flexible, adaptable and able to navigate the ever-changing environment. This fast-paced environment has however resulted in an organizational landscape that battles to engage employees, retain top talent and create meaningful work for its members. In the knowledge economy, the concept of organizational identity has become an important consideration as organisations aim to create a compelling and inviting narrative for all stakeholders across the business value chain. Values are often seen as the behavioural framework that informs organisational culture, yet often values are perceived to be inauthentic and misaligned with the true character or identity of the organisation and how it is perceived by different role players. This paper focuses on exploring the relationship between organisational identity and value systems by focusing on a case study within a multi-national organisation within South Africa. The paper evaluates the implementation of mixed methods OD approach that gathered collaborative inputs of more than 4500 employees who participated in crafting the newly established values system post a retrenchment process. The paper will evaluate the relationship between the newly crafted value system and the identity of the organisation as described by various internal and external stakeholders in order to explore potential alignment, dissonance and key insights into understanding the relationship between organisational identity and values. The case study will be reported from the perspective of an OD consultant who supported the transformation process over a period of 8 months and aims to provide key insights into values and identity alignment within knowledge economy organisations. From a practical perspective, the paper provides insights into how values are created, perceived and lived within organisations and the impact on employee engagement and culture.

Keywords: culture, organisational development, organisational identity, values

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Authors: Marina Sartor Hoffer

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The multicultural composition of today's societies poses new challenges to educational contexts. Schools are therefore called first to develop dialogic aptitudes and communicative skills adapted to the complex reality of post-modern societies. It is indispensable for educators and for young people to learn theoretical and practical tools during their scholastic path, in order to allow the knowledge of themselves and of the others with the aim of recognizing the value of the others regardless of their culture. Dialogic Skills help to understand and manage individual differences by allowing the solution of problems and preventing conflicts. The Educational Sector of Eric Berne’s Transactional Analysis offers a range of methods and techniques for this purpose. Educational Transactional Analysis is firmly anchored in the Personalist Philosophy and deserves to be promoted as a theoretical frame suitable to face the challenges of contemporary education. The goal of this paper is therefore to outline some conceptual and methodological signposts for the education to dialogue by drawing concepts and methodologies from educational transactional analysis.

Keywords: dialogic process, education to dialogue, educational transactional analysis, personalism, the good of the relationship

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Authors: Martina Depentor

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While the cinematic medium contributes to bringing images of a culture to foreign audiences, Audiovisual Translation contributes to deciphering those cultural representations to those same audiences. Through Audiovisual Translation, in fact, elements permeate the reception system and contribute to forging a cultural image of the original/source system in the target/reception system. By analyzing a number of Italian critical reviews, blogs and forum posts, this paper examines the impact and reception in Italy of five of the most successful and influential New Zealand films of the last two decades - An Angel at my Table (1990), The Piano (1993), Heavenly Creatures (1994), Once Were Warriors (1994), Whale Rider (2002) - with the aim of exploring how the adaptation of New Zealand films might condition the representation of New Zealand in the Italian imaginary. The analysis seeks to identify whether a certain degree of cultural loss results from the 'translation' of these films. The films selected share common ground in that they all reveal cultural, social and historical characteristics of New Zealand, from aspects that are unique to this country and that on the surface may render it difficult to penetrate (unfamiliar landscapes, aspects of indigenous culture) to more universal themes (intimate family stories, dysfunctional relationship). They contributed to situating New Zealand on an international stage and to bringing images of the country to many audiences, the Italian one included, with little previous cultural knowledge of the social and political history of New Zealand. Differences in film types pose clearly different levels of interpretative challenges to non-New Zealander audiences, and examples from the films will show how these challenges are or are not overcome if the adaptations display misinterpretations or rendition gaps, and how the process of intercultural transfer further 'domesticates' or 'exoticises' the source culture.

Keywords: audiovisual translation, cultural representation, intercultural transfer, New Zealand Films

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Authors: Galina Mihaleva, C. Koh

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Fashion, like many other design fields, undergoes numerous evolutions throughout the ages. This paper aims to recognize and evaluate the significance of advance technology in fashion design and examine how it changes the role of modern fashion designers by modifying the creation process. It also touches on how modern culture is involved in such developments and how it affects fashion design in terms of conceptualizing and fabrication. The methodology used is through surveying the various examples of technological applications to fashion design and drawing parallels between what was achievable then and what is achievable now. By comparing case studies, existing fashion design examples and crafting method experimentations; we then spot patterns in which to predict the direction of future developments in the field. A breakdown on the elements of technology in fashion design helps us understand the driving force behind such a trend. The results from explorations in the paper have shown that there is an observed pattern of a distinct increase in interest and progress in the field of fashion technology, which leads to the birth of hybrid crafting methods. In conclusion, it is shown that as fashion technology continues to evolve, their role in clothing crafting becomes more prominent and grows far beyond the humble sewing machine.

Keywords: fashion design, functional aesthetics, smart textiles, 3D printing

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Authors: Patrícia Branco, Catarina Prista, Helena Albergaria

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Industrial fuel-ethanol fermentations are carried out under non-sterile conditions, which favors the development of microbial contaminants, leading to huge economic losses. Wild yeasts such as Brettanomyces bruxellensis and lactic acid bacteria are the main contaminants of industrial bioethanol fermentation, affecting Saccharomyces cerevisiae performance and decreasing ethanol yields and productivity. In order to control microbial contaminations, the fuel-ethanol industry uses different treatments, including acid washing and antibiotics. However, these control measures carry environmental risks such as acid toxicity and the rise of antibiotic-resistant bacteria. Therefore, it is crucial to develop and apply less toxic and more environmentally friendly biocontrol methods. In the present study, an industrial fuel-ethanol starter, S. cerevisiae Ethanol-Red, was genetically modified to over-express AMPs with activity against fuel-ethanol microbial contaminants and evaluated regarding its biocontrol effect during mixed-culture alcoholic fermentations artificially contaminated with B. bruxellensis. To achieve this goal, S. cerevisiae Ethanol-Red strain was transformed with a plasmid containing the AMPs-codifying genes, i.e., partial sequences of TDH1 (925-963 bp) and TDH2/3 (925-963 bp) and a geneticin resistance marker. The biocontrol effect of those genetically modified strains was evaluated against B. bruxellensis and compared with the antagonistic effect exerted by the modified strain with an empty plasmid (without the AMPs-codifying genes) and the non-modified strain S. cerevisiae Ethanol-Red. For that purpose, mixed-culture alcoholic fermentations were performed in a synthetic must use the modified S. cerevisiae Ethanol-Red strains together with B. bruxellensis. Single-culture fermentations of B. bruxellensis strains were also performed as a negative control of the antagonistic effect exerted by S. cerevisiae strains. Results clearly showed an improved biocontrol effect of the genetically-modified strains against B. bruxellensis when compared with the modified Ethanol-Red strain with the empty plasmid (without the AMPs-codifying genes) and with the non-modified Ethanol-Red strain. In mixed-culture fermentation with the modified S. cerevisiae strain, B. bruxellensis culturability decreased from 5×104 CFU/mL on day-0 to less than 1 CFU/mL on day-10, while in single-culture B. bruxellensis increased its culturability from 6×104 to 1×106 CFU/mL in the first 6 days and kept this value until day-10. Besides, the modified Ethanol-Red strain exhibited an enhanced antagonistic effect against B. bruxellensis when compared with that induced by the non-modified Ethanol-Red strain. Indeed, culturability loss of B. bruxellensis after 10 days of fermentation with the modified Ethanol-Red strain was 98.7 and 100% higher than that occurred in fermentations performed with the non-modified Ethanol-Red and the empty-plasmid modified strain, respectively. Therefore, one can conclude that the S. cerevisiae genetically modified strain obtained in the present work may be a valuable solution for the mitigation of microbial contamination in fuel-ethanol fermentations, representing a much safer and environmentally friendly preservation strategy than the antimicrobial treatments (acid washing and antibiotics) currently applied in fuel-ethanol industry.

Keywords: antimicrobial peptides, fuel-ethanol microbial contaminations, fuel-ethanol fermentation, biocontrol agents, genetically-modified yeasts

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Authors: Atje Setiawan Abdullah, Budi Nurani Ruchjana, I. Gede Nyoman Mindra Jaya, Eddy Hermawan

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Ethnoscience is used to see the culture with a scientific perspective, which may help to understand how people develop various forms of knowledge and belief, initially focusing on the ecology and history of the contributions that have been there. One of the areas studied in ethnoscience is etno-informatics, is the application of informatics in the culture. In this study the science of informatics used is data mining, a process to automatically extract knowledge from large databases, to obtain interesting patterns in order to obtain a knowledge. While the application of culture described by naming database village on the island of Java were obtained from Geographic Indonesia Information Agency (BIG), 2014. The purpose of this study is; first, to classify the naming of the village on the island of Java based on the structure of the word naming the village, including the prefix of the word, syllable contained, and complete word. Second to classify the meaning of naming the village based on specific categories, as well as its role in the community behavioral characteristics. Third, how to visualize the naming of the village to a map location, to see the similarity of naming villages in each province. In this research we have developed two theorems, i.e theorems area as a result of research studies have collected intersection naming villages in each province on the island of Java, and the composition of the wedge theorem sets the provinces in Java is used to view the peculiarities of a location study. The methodology in this study base on the method of Knowledge Discovery in Database (KDD) on data mining, the process includes preprocessing, data mining and post processing. The results showed that the Java community prioritizes merit in running his life, always working hard to achieve a more prosperous life, and love as well as water and environmental sustainment. Naming villages in each location adjacent province has a high degree of similarity, and influence each other. Cultural similarities in the province of Central Java, East Java and West Java-Banten have a high similarity, whereas in Jakarta-Yogyakarta has a low similarity. This research resulted in the cultural character of communities within the meaning of the naming of the village on the island of Java, this character is expected to serve as a guide in the behavior of people's daily life on the island of Java.

Keywords: ethnoscience, ethno-informatics, data mining, clustering, Java island culture

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Authors: Swati Kapoor, Rajeev Upreti, Monica Mahajan, Abhaya Indrayan, Dinesh Srivastava

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Enteric Fever is caused by gram negative bacilli Salmonella typhi and paratyphi. It is associated with high morbidity and mortality worldwide. Timely initiation of treatment is a crucial step for prevention of any complications. Cultures of body fluids are diagnostic, but not always conclusive or practically feasible in most centers. Moreover, the results of cultures delay the treatment initiation. Serological tests lack diagnostic value. The blood counts can offer a promising option in diagnosis. A retrospective study to find out the relevance of leucopenia and eosinopenia was conducted on 203 culture proven enteric fever patients and 159 culture proven non-enteric fever patients in a tertiary care hospital in New Delhi. The patient details were retrieved from the electronic medical records section of the hospital. Absolute eosinopenia was considered as absolute eosinophil count (AEC) of less than 40/mm³ (normal level: 40-400/mm³) using LH-750 Beckman Coulter Automated machine. Leucopoenia was defined as total leucocyte count (TLC) of less than 4 X 10⁹/l. Blood cultures were done using BacT/ALERT FA plus automated blood culture system before first antibiotic dose was given. Case and control groups were compared using Pearson Chi square test. It was observed that absolute eosinophil count (AEC) of 0-19/mm³ was a significant finding (p < 0.001) in enteric fever patients, whereas leucopenia was not a significant finding (p=0.096). Using Receiving Operating Characteristic (ROC) curves, it was observed that patients with both AEC < 14/mm³ and TCL < 8 x 10⁹/l had 95.6% chance of being diagnosed as enteric fever and only 4.4% chance of being diagnosed as non-enteric fever. This result was highly significant with p < 0.001. This is a very useful association of AEC and TLC found in enteric fever patients of this study which can be used for the early initiation of treatment in clinically suspected enteric fever patients.

Keywords: absolute eosinopenia, absolute eosinophil count, enteric fever, leucopenia, total leucocyte count

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Authors: Maged El-Sayed Mohamed

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Plant tissue culture practices are presented nowadays as the most promising substitute to a whole plant in the terms of secondary metabolites production. They offer the advantages of high production, tunability and they have less effect on plant ecosystems. Lantana camara is a weed, which is common all over the world as an ornamental plant. Weeds can adapt to any type of soil and climate due to their rich cellular machinery for secondary metabolites’ production. This characteristic is found in Lantana camara as a plant of very rich diversity of secondary metabolites with no dominant class of compounds. Aim: This trait has encouraged the author to develop tissue culture experiments for Lantana camara to be a platform for production and manipulation of secondary metabolites through biotransformation. Methodology: The plant was collected in its flowering stage in September 2014, from which explants were prepared from shoot tip, auxiliary bud and leaf. Different types of culture media were tried as well as four phytohormones and their combinations; NAA, 2,4-D, BAP and kinetin. Explants were grown in dark or in 12 hours dark and light cycles at 25°C. A metabolic profile for the produced callus was made and then compared to the whole plant profile. The metabolic profile was made using GC-MS for volatile constituents (extracted by n-hexane) and by HPLC-MS and capillary electrophoresis-mass spectrometry (CE-MS) for non-volatile constituents (extracted by ethanol and water). Results: The best conditions for the callus induction was achieved using MS media supplied with 30 gm sucrose and NAA/BAP (1:0.2 mg/L). Initiation of callus was favoured by incubation in dark for 20 day. The callus produced under these conditions showed yellow colour, which changed to brownish after 30 days. The rate of callus growth was high, expressed in the callus diameter, which reached to 1.15±0.2 cm in 30 days; however, the induction of callus delayed for 15 days. The metabolic profile for both volatile and non-volatile constituents of callus showed more simple background metabolites than the whole plant with two new (unresolved) peaks in the callus’ nonvolatile constituents’ chromatogram. Conclusion: Lantana camara callus production can be itself a source of new secondary metabolites and could be used for biotransformation studies due to its simple metabolic background, which allow easy identification of newly formed metabolites. The callus production gathered the simple metabolic background with the rich cellular secondary metabolite machinery of the plant, which could be elicited to produce valuable medicinally active products.

Keywords: capillary electrophoresis-mass spectrometry, gas chromatography, metabolic profile, plant tissue culture

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Authors: William Kelly, Sorelle Veigne, Xianhua Li, Zuyi Huang, Shyamsundar Subramanian, Eugene Schaefer

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The ambr15™ bioreactor is a single-use microbioreactor for cell line development and process optimization. The ambr system offers fully automatic liquid handling with the possibility of fed-batch operation and automatic control of pH and oxygen delivery. With operating conditions for large scale biopharmaceutical production properly scaled down, micro bioreactors such as the ambr15™ can potentially be used to predict the effect of process changes such as modified media or different cell lines. In this study, gassing rates and dilution rates were varied for a semi-continuous cell culture system in the ambr15™ bioreactor. The corresponding changes to metabolite production and consumption, as well as cell growth rate and therapeutic protein production were measured. Conditions were identified in the ambr15™ bioreactor that produced metabolic shifts and specific metabolic and protein production rates also seen in the corresponding larger (5 liter) scale perfusion process. A Dynamic Flux Balance model was employed to understand and predict the metabolic changes observed. The DFB model-predicted trends observed experimentally, including lower specific glucose consumption when CO₂ was maintained at higher levels (i.e. 100 mm Hg) in the broth. A Computational Fluid Dynamic (CFD) model of the ambr15™ was also developed, to understand transfer of O₂ and CO₂ to the liquid. This CFD model predicted gas-liquid flow in the bioreactor using the ANSYS software. The two-phase flow equations were solved via an Eulerian method, with population balance equations tracking the size of the gas bubbles resulting from breakage and coalescence. Reasonable results were obtained in that the Carbon Dioxide mass transfer coefficient (kLa) and the air hold up increased with higher gas flow rate. Volume-averaged kLa values at 500 RPM increased as the gas flow rate was doubled and matched experimentally determined values. These results form a solid basis for optimizing the ambr15™, using both CFD and FBA modelling approaches together, for use in microscale simulations of larger scale cell culture processes.

Keywords: cell culture, computational fluid dynamics, dynamic flux balance analysis, microbioreactor

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Authors: Utsha Barua Joy

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Buddhism has been playing as a cultural mediator in the world Asia. Buddhism spread as a living spiritual tradition and philosophy and spread mostly as a part of the cultural heritage. Culture and religion concern and deal with some similar human issues in terms of the concept of values. This article mainly focuses on some pertinent issues. The Buddha created a revolutionary awareness in the history of mankind by giving equal status to all. With equality, man gets social, mental, economic, and political freedom. This article moreover discusses the concept of morality, educational system, and finally, missionary services. After the first rainy retreat, Buddha asked the monks to go from place to place and spread the Dharma for the welfare of mankind. Since then, all the monks took part in missionary work. Emperor Asoka’s missionary steps are the brightened example in the history of Buddhism. The goal of this article is to provide to the wise readers with a delineation of reference on how equality, morality, education system, and missionary services through Buddhism rendered great contribution to the cultural arena around the globe.

Keywords: equality, morality, educational system, missionary works

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Authors: Ka Lei Au

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The Dalcroze method has been emerging in music classrooms, and music teachers are encouraged to integrate music and movement in their teaching. Music programs in colleges in Hong Kong have been introducing method courses such as Orff and Dalcroze method in music teaching as teacher’s education program. Since the targeted students of the course are music teachers who are making the decision of what approach to use in their classroom, their perception is significantly valued to identify how this approach is applicable in their teaching in regards to the teaching and learning culture and environment. This qualitative study aims to explore how the Dalcroze method as a teacher’s education course is perceived by music teachers from three aspects: 1) application in music teaching, 2) self-enhancement, 3) expectation. Through the lens of music teachers, data were collected from 30 music teachers who are taking the Dalcroze method course in music teaching in Hong Kong by the survey. The findings reveal the value and their intention of the Dalcroze method in Hong Kong. It also provides a significant reference for better development of such courses in the future in adaption to the culture, teaching and learning environment and teacher’s, student’s and parent’s perception of this approach.

Keywords: Dalcroze method, music teaching, perception, self-enhancement, teacher’s education

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Authors: Quin Emparan, Razif Harun, Dayang R. A. Biak, Rozita Omar, Michael K. Danquah

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Cultivation of immobilized microalgae cells is on the rise for biotechnological applications. In this study, cultivation of Chlorella vulgaris was carried out in the form of suspended free-cell and immobilized cells system. NaCS-PDMDAAC capsules were used to immobilize C. vulgaris. Initially, the synthesized NaCS with C. vulgaris culture were prepared at various concentration of 5- 20% (w/v) using a 6% hardening solution (PDMDAAC) to investigate the capsules' gel stability and suitability for microalgae cells growth. Then, the capsules produced from 15% NaCS with C. vulgaris culture were furthered investigated using 5%, 10%, and 15% (w/v) of PDMDAAC solution. The capsules' gel stability was evaluated through dissolution time and loss of uniform spherical shape of capsules, while suitability for microalgae cells growth was evaluated through the optical density of microalgae. In this study, the 15% NaCS-10% PDMDAAC capsules were found to be the most suitable to sustain the capsules' gel stability and microalgae cells growth in MLA. For that reason, the C. vulgaris immobilized in the 15% NaCS-10% PDMDAAC capsules were further characterized using physicochemical analysis in terms of morphological, carbon (C), hydrogen (H) and nitrogen (N), Fourier transform-infrared (FT-IR), scanning electron microscopy-energy dispersive X-ray (SEM-EDX), zeta potential and Brunauer-Emmet-Teller (BET) analyses. The results revealed that the presence of sulfonates in the synthesized NaCS and NaCS-PDMDAAC capsules without and with C. vulgaris proves that cellulose alcohol group was successfully bonded by sulfo group. Besides that, immobilized microalgae cells have a smaller cell size of 6.29 ± 1.09 µm and zeta potential of -11.93 ± 0.91 mV than suspended free-cells microalgae culture. It can be summarized that immobilization of C. vulgaris in the 15% NaCS-10% PDMDAAC capsules are relevant as a bioremediator for wastewater treatment purposes due to its suitable size of pore and capsules as well as structural and compositional properties.

Keywords: biological capsules, immobilized cultivation, microalgae, physico-chemical analysis

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Authors: Muhammad Suhail Ibrahim, Ahmad Mujtaba

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Background: The accuracy of the most frequently used tests for diagnosing Helicobacter pylori is always under consideration in clinical settings. A reliable diagnosis is crucial to confirm the success of therapy. Objective: The aim of this research was to study the isolation frequency of H. pylori from patients compatible with gastritis or gastric ulcer and to compare some feasible non-invasive and invasive methods for the diagnosis of infection. Materials and Methods: Ninety-six gastric biopsy and blood samples were obtained with various gastroduodenal symptoms after obtaining informed consent. The biopsies were analyzed and compared using the culture, microscopic examination, histopathology, Rapid urease RUT), serology, biochemical, antibiotic susceptibility test and molecular method. Results: A number of 40 (41.67%) were considered H. pylori positive in both histopathology and RUT. On the other hand, 46 patients were positive against anti IgA and IgG by ELISA. Eighteen biopsies were positive according to the culture test. This was further confirmed by endoscopic examination, urease, catalase and oxidase tests. A high percentage of resistance to polymyxin B, amoxicillin, and kanamycin was observed (100, 88.89, and 77.78%, respectively). A gene (Cag A) was also detected by using molecular technique which appeared positive in 16 patients. The sensitivity/specificity (%) of diagnostic method was 95/77 for histology, 100/83.5 for rapid urease, 85.7/90 for gram staining, 100/66.6 for IgG serology, 100/79.5 for IgA serology, 100/75.0 for PCR, 100/79.04 for combination of RUT and IgG serology and 100/92.4 for combination of RUT, gram staining and IgG serology. Conclusion: In view of the result obtained, PCR appeared to be the most reliable test. However, higher sensitivity and specificity were also recorded for other tests. So, for more accurate results, it is advisable not to rely solely on a single method for detection.

Keywords: helicobacter pylori, isolation, detection, culture, urease, polymerase chain reaction, antibiotic susceptibility test, dyspeptic patients

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Authors: Jarot Mahendra

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The primary objective of this paper is to provide a view in the interpretation of the revitalization of heritage buildings. This objective is achieved by analyzing the concept of interpretation that is oriented in the perspective of law, urban spatial planning, and stakeholder perspective, and then develops the theoretical framework of interpretation in the cultural resources management through issues of identity, heritage as a process, and authenticity in heritage. The revitalization of heritage buildings with the interpretation of these three issues is that interpretation can be used as a communication process to express the meaning and relation of heritage to the community so as to avoid the conflict that will arise and develop as a result of different perspectives of stakeholders. Using case studies in Indonesia, this study focuses on the revitalization of heritage sites in the National Gallery of Indonesia (GNI). GNI is a cultural institution that uses several historical buildings that have been designated as heritage and have not been designated as a heritage according to the regulations applicable in Indonesia, in carrying out its function as the center of Indonesian art development and art museums. The revitalization of heritage buildings is taken as a step to meet space needs in running the current GNI function. In the revitalization master plan, there are physical interventions on the building of heritage and the removal of some historic buildings which will then be built new buildings at that location. The research matrix was used to map out the main elements of the study (the concept of GNI revitalization, heritage as identity, heritage as a process, and authenticity in the heritage). Expert interviews and document studies are the main tools used in collecting data. Qualitative data is then analyzed through content analysis and template analysis. This study identifies the significance of historic buildings (heritage buildings and buildings not defined as heritage) as an important value of history, architecture, education, and culture. The significance becomes the basis for revisiting the revitalization master plan which is then reviewed according to applicable regulations and the spatial layout of Jakarta. The interpretation that is built is (1) GNI is one of the elements of the embodiment of the National Cultural Center in the context of the region, where there are National Monument, National Museum and National Library in the same area, so the heritage not only gives identity to the past culture but the culture of current community; (2) The heritage should be seen as a dynamic cultural process towards the cultural change of community, where heritage must develop along with the urban development, so that the heritage buildings can remain alive and side by side with modern buildings but still observe the principles of preservation of heritage; (3) The authenticity of heritage should be able to balance the cultural heritage conservation approach with urban development, where authenticity can serve as a 'Value Transmitter' so that authenticity can be used to evaluate, preserve and manage heritage buildings by considering tangible and intangible aspects.

Keywords: authenticity, culture process, identity, interpretation, revitalization

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Authors: Eva Tvrdá, Boris Botman, Marek Halenár, Tomáš Slanina, Norbert Lukáč

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In vitro storage and processing of animal semen represents a risk factor to spermatozoa vitality, potentially leading to reduced fertility. A variety of substances isolated from natural sources may exhibit protective or antioxidant properties on the spermatozoon, thus extending the lifespan of stored ejaculates. This study compared the ability of different concentrations of the Salvia officinalis extract on the motility, mitochondrial activity, viability and reactive oxygen species (ROS) production by bovine spermatozoa during different time periods (0, 2, 6 and 24 h) of in vitro culture. Spermatozoa motility was assessed using the Computer-assisted sperm analysis (CASA) system. Cell viability was examined using the metabolic activity MTT assay, the eosin-nigrosin staining technique was used to evaluate the sperm viability and ROS generation was quantified using luminometry. The CASA analysis revealed that the motility in the experimental groups supplemented with 0.5-2 µg/mL Salvia extract was significantly lower in comparison with the control (P<0.05; Time 24 h). At the same time, a long-term exposure of spermatozoa to concentrations ranging between 0.05 µg/mL and 2 µg/mL had a negative impact on the mitochondrial metabolism (P<0.05; Time 24 h). The viability staining revealed that 0.001-1 µg/mL Salvia extract had no effects on bovine male gametes, however 2 µg/mL Salvia had a persisting negative effect on spermatozoa (P<0.05). Furthermore 0.05-2 µg/mL Salvia exhibited an immediate ROS-promoting effect on the sperm culture (P>0.05; Time 0 h and 2 h), which remained significant throughout the entire in vitro culture (P<0.05; Time 24 h). Our results point out to the necessity to examine specific effects the biomolecules present in Salvia officinalis may have individually or collectively on the in vitro sperm vitality and oxidative profile.

Keywords: bulls, CASA, MTT test, reactive oxygen species, sage, Salvia officinalis, spermatozoa

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Authors: Antonello Durante, James Duff, Kevin Kelly

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The visual needs of people in modern work based buildings are changing. Self-illuminated screens of computers, televisions, tablets and smart phones have changed the relationship between people and the lit environment. In the past, lighting design practice was primarily based on providing uniform horizontal illuminance on the working plane, but this has failed to ensure good quality lit environments. Lighting standards of today continue to be set based upon a 100 year old approach that at its core, considers the task illuminance of the utmost importance, with this task typically being located on a horizontal plane. An alternative method focused on appearance has been proposed, as opposed to the traditional performance based approach. Mean Room Surface Exitance (MRSE) and Target-Ambient Illuminance Ratio (TAIR) are two new metrics proposed to assess illumination adequacy in interiors. The hypothesis is that these factors will be superior to the existing metrics used, which are horizontal illuminance led. For the six past years, research has examined this, within the Dublin Institute of Technology, with a view to determining the suitability of this approach for application to general lighting practice. Since the start of this research, a number of key findings have been produced that centered on how occupants will react to various levels of MRSE. This paper provides a broad update on how this research has progressed. More specifically, this paper will: i) Demonstrate how MRSE can be measured using HDR images technology, ii) Illustrate how MRSE can be calculated using scripting and an open source lighting computation engine, iii) Describe experimental results that demonstrate how occupants have reacted to various levels of MRSE within experimental office environments.

Keywords: illumination hierarchy (IH), mean room surface exitance (MRSE), perceived adequacy of illumination (PAI), target-ambient illumination ratio (TAIR)

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Authors: Lin Chih-Ken

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This paper explores the A Li Shan hotel of Taiwan during the Japanese occupation period, after over a hundred years of time, it has been handed over to the hotel managing enterprise to retain the historic building and the culture. Applying the innovative marketing strategies, coordinate the local government traveling policy then combined local tea agriculture and forestry specialty integrated marketing, to create the special hotel located in the Alishan National Scenic Area with the characteristics of landscape, innovative marketing and history, to attract domestic tourism and visitors around the world. This study interview the hotel owner, managers, employees and guests, in addition to collected message feedback from reservation website, to apply Ambidexterity Marketing Theory and Resource Base Theory to analyze the main impact factors. The conclusion showed that the integration of several key factors and make good use of resource strength generate heterogeneous product characteristics to attracting wider range of visitors.

Keywords: heritage tourism, historic hotel, marketing ambidexterity, resource base theory

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Authors: Waad Al Saleemi, Badriya Al Adawi, Zaaima Al Jabri, Sahim Al Ghafri, Jalila Al Hadhramia

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Objectives: Using standard lab methods, a positive blood culture requires a minimum of two days (two occasions of overnight incubation) to obtain a final identification (ID) and antimicrobial susceptibility results (AST) report. In this study, we aimed to evaluate the accuracy and precision of identification and antimicrobial susceptibility testing of an alternative method (direct method) that will reduce the turnaround time by 24 hours. This method involves the direct inoculation of positive blood culture broths into the Phoenix system using serum separation tubes (SST). Method: This prospective study included monomicrobial-positive blood cultures obtained from January 2022 to May 2023 in SQUH. Blood cultures containing a mixture of organisms, fungi, or anaerobic organisms were excluded from this study. The result of the new “direct method” under study was compared with the current “standard method” used in the lab. The accuracy and precision were evaluated for the ID and AST using Clinical and Laboratory Standards Institute (CLSI) recommendations. The categorical agreement, essential agreement, and the rates of very major errors (VME), major errors (ME), and minor errors (MIE) for both gram-negative and gram-positive bacteria were calculated. Passing criteria were set according to CLSI. Result: The results of ID and AST were available for a total of 158 isolates. Of 77 isolates of gram-negative bacteria, 71 (92%) were correctly identified at the species level. Of 70 isolates of gram-positive bacteria, 47(67%) isolates were correctly identified. For gram-negative bacteria, the essential agreement of the direct method was ≥92% when compared to the standard method, while the categorical agreement was ≥91% for all tested antibiotics. The precision of ID and AST were noted to be 100% for all tested isolates. For gram-positive bacteria, the essential agreement was >93%, while the categorical agreement was >92% for all tested antibiotics except moxifloxacin. Many antibiotics were noted to have an unacceptable higher rate of very major errors including penicillin, cotrimoxazole, clindamycin, ciprofloxacin, and moxifloxacin. However, no error was observed in the results of vancomycin, linezolid, and daptomycin. Conclusion: The direct method of ID and AST for positive blood cultures using SST is reliable for gram negative bacteria. It will significantly decrease the turnaround time and will facilitate antimicrobial stewardship.

Keywords: bloodstream infection, oman, direct ast, blood culture, rapid identification, antimicrobial susceptibility, phoenix, direct inoculation

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Authors: Ruchi Agarwal

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In today’s world, companies are operating in dynamic, uncertain and ambiguous business environments. Globally, more companies are failing due to Environmental, Social and Governance (ESG) factors than ever. Corporate governance and risk management are intertwined in nature. For decades, corporate governance and risk management have been influenced by internal and external factors. Three schools of thought have influenced risk governance for decades: Agency theory, Contingency theory, and Institutional theory. Agency theory argues that agents have interests conflicting with principal interests and the information problem. Contingency theory suggests that risk management adoption is influenced by internal and external factors, while Institutional theory suggests that organizations legitimize risk management with regulators, competitors, and professional bodies. The conflicting objectives of theories have created problems for executives in organizations in the adoption of Risk Governance. So far, there are many studies that discussed risk culture and the role of actors in risk governance, but there are rare studies discussing the role of risk culture in the adoption of risk governance from a leadership style perspective. This study explores the adoption of risk governance in two contrasting industries, such as the Insurance and energy business, to understand whether risk governance is influenced by internal/external factors or whether risk culture is influenced by leaders. We draw empirical evidence by comparing the cases of an Indian insurance company and a renewable energy-based firm in India. We interviewed more than 20 senior executives of companies and collected annual reports, risk management policies, and more than 10 PPTs and other reports from 2017 to 2024. We visited the company for follow-up questions several times. The findings of my research revealed that both companies have used risk governance for strategic renewal of the company. Insurance companies use a transactional leadership style based on performance and reward for improving risk, while energy companies use rather symbolic management to make debt restructuring meaningful for stakeholders. Overall, both companies turned from loss-making to profitable ones in a few years. This comparative study highlights the role of different leadership styles in the adoption of risk governance. The study is also distinct as previous research rarely studied risk governance in two contrasting industries in reference to leadership styles.

Keywords: leadership style, corporate governance, risk management, risk culture, strategic renewal

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Authors: Daniele Abreu e Lima, Chloe Fitzpatrick

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During the 1930s, New Zealand received a wave of refugees feeling from the impeding war and atrocities the Nazi regime was imposing on the German people. Among the hundreds of refugees were highly trained artists, architects and musicians who made a huge contribution to Wellington’s culture and identity. It is unfeasible to chronicle the impact of every Jewish refugee in the development of New Zealand arts scene. But it is possible to choose a number of them and analyse their contribution to NZ culture. This research aims to bring to light the reception and life of five influential Jewish architects; Helmut Einhorn, Ernst Plischke, Frederick Neumann, Henry Kulka, and Maximillian Rosenfeld. Each had a key role in influencing New Zealand architectural landscape and the modernization of the country. Before coming to New Zealand, these five architects lived different lives working all over Europe, from Paris through to Moscow. In common, apart from their ethnicity, they had led cultured lives where they were culturally and politically active. This research looks at how much their individual contributions helped to transform the architectural scene in New Zealand but also in the amount of cultural and religious renunciation they had to endure to be accepted in the country.

Keywords: Jewish Refugee architects, modern architecture, World War 2, New Zealand

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Authors: Melissa Valle

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This paper draws upon the studies of visual culture and intersectionality to illuminate how visuality can naturalize social hierarchies. Through the use of iconography, it decodes the denotative, connotative and ideological meanings of symbols of ritualistic events in the context of the Colombian Atlantic Coast. An examination of such exceptional moments, i.e. of the spectacle, brings into focus how such performances are imbued with meaning by both the on-looker and the performer. Through an analysis of preexisting visuals (e.g., advertisements, social media) and visual materials produced by the researcher for the purpose of photo-elicitation interviews, this paper provides a contextual analysis of the ways in which three representations, popular during Colombian Atlantic coastal festivals (Negrita Puloy, Las Palenqueras, and El Son de Negro), have been historically, culturally and politically constituted. This work reveals that the visualizations are born out of and reproduce typifications systems heavily based upon race, gender, class, and ethnicity. Understanding the ways these categories are mutually constituted through the cultural practice of visual representation is essential to a more comprehensive understanding of the role such representation plays in the reproduction of social difference.

Keywords: Colombia, festivals, intersectionality, visual culture

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Authors: Demur Bakuradze, Mzia Beruashvili

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It is known that bioactive compounds that are secreted by allelopathic plants sown with the main culture and are natural phytotoxic substances can be used as natural pesticides that, unlike synthetic agrochemicals, are less likely to disrupt the global ecosystem. Alternative plant protection approaches to date include components of integrated pest management programs and natural plant protection tools. Every year, more and more evidence is given on the successful use of these means, including allelopathic compounds in plant protection against pests. Proper use of these interactions between plants and other organisms can become a serious alternative to synthetic pesticides. Due to the relevance of the issue, the purpose of the study was to study the impact of some companion allelopathic plants on the Tomato culture (variety ‘Kharisgula’) on the test plot of the Tsilkani Base Bioagricultural Service of the Agricultural Research Center (East Georgia) and the village of Kharagauli. The study was conducted in 2023-2024 with methods adopted in organic agriculture and plant protection. Various companion plants were studied: Basil (Ocimum basilicum L.), Imeretian saffron or marigold (Tagetes patula L.) and beans (Phaseolus vulgaris L.) (variety ‘Tsanava’) affects tomato culture in mixed crops. It was established that when sowing basil in rows and on the perimeter, the yield in the Tsilkani increased by 39.7%, and in Sagandzile - 42.8%. In the case of Imeretian saffron, economic efficiency in Tsilkani reached 29.5%, in Sagandzile - 28.5%, while the difference in the bean variant with the control was 31.7% in Tsilkani, and in Sagandzile - 23.8%. The study also found that when sowing aromatic plants (basil, marigold) with tomato, the number and spread of pests is quite decreasing compared to the control. In particular, the number of green vegetable bugs (Nezara viridula L.) decreased by 59.4% compared to control.

Keywords: allelopathy, companion planting, tomato, bioproduction

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Authors: E. Nakamachi, S. Tanaka, K. Yamamoto, Y. Morita

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In this study, we developed a three-dimensional (3D) direct current electric field (DCEF) stimulation bio-reactor for axonal outgrowth enhancement to generate the neural network of the central nervous system (CNS). By using our newly developed 3D DCEF stimulation bio-reactor, we cultured the rat pheochromocytoma cells (PC12) and investigated the effects on the axonal extension enhancement and network generation. Firstly, we designed and fabricated a 3D bio-reactor, which can load DCEF stimulation on PC12 cells embedded in the collagen gel as extracellular environment. The connection between the electrolyte and the medium using salt bridges for DCEF stimulation was introduced to avoid the cell death by the toxicity of metal ion. The distance between the salt bridges was adopted as the design variable to optimize a structure for uniform DCEF stimulation, where the finite element (FE) analyses results were used. Uniform DCEF strength and electric flux vector direction in the PC12 cells embedded in collagen gel were examined through measurements of the fabricated 3D bio-reactor chamber. Measurement results of DCEF strength in the bio-reactor showed a good agreement with FE results. In addition, the perfusion system was attached to maintain pH 7.2 ~ 7.6 of the medium because pH change was caused by DCEF stimulation loading. Secondly, we disseminated PC12 cells in collagen gel and carried out 3D culture. Finally, we measured the morphology of PC12 cell bodies and neurites by the multiphoton excitation fluorescence microscope (MPM). The effectiveness of DCEF stimulation to enhance the axonal outgrowth and the neural network generation was investigated. We confirmed that both an increase of mean axonal length and axogenesis rate of PC12, which have been exposed 5 mV/mm for 6 hours a day for 4 days in the bioreactor. We found following conclusions in our study. 1) Design and fabrication of DCEF stimulation bio-reactor capable of 3D culture nerve cell were completed. A uniform electric field strength of average value of 17 mV/mm within the 1.2% error range was confirmed by using FE analyses, after the structure determination through the optimization process. In addition, we attached a perfusion system capable of suppressing the pH change of the culture solution due to DCEF stimulation loading. 2) Evaluation of DCEF stimulation effects on PC12 cell activity was executed. The 3D culture of PC 12 was carried out adopting the embedding culture method using collagen gel as a scaffold for four days under the condition of 5.0 mV/mm and 10mV/mm. There was a significant effect on the enhancement of axonal extension, as 11.3% increase in an average length, and the increase of axogenesis rate. On the other hand, no effects on the orientation of axon against the DCEF flux direction was observed. Further, the network generation was enhanced to connect longer distance between the target neighbor cells by DCEF stimulation.

Keywords: PC12, DCEF stimulation, 3D bio-reactor, axonal extension, neural network generation

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