Search results for: mRNA expression

Authors: Maryam Parsian, Pelin Mutlu, Ufuk Gunduz

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Two-dimensional cell culture affords simplicity and low cost, but it has serious limitations; lacking cell-cell and cell-matrix interactions that are present in tissues. Cancer cells grown in 3D culture systems have distinct phenotypes of adhesion, growth, migration, invasion as well as profiles of gene and protein expression. These interactions cause the 3D-cultured cells to acquire morphological and cellular characteristics relevant to in vivo tumors. Palbociclib is a chemotherapeutic agent for the treatment of ER-positive and HER-negative metastatic breast cancer. Poly-amidoamine (PAMAM) dendrimer is a well-defined, special three-dimensional structure and has a multivalent surface and internal cavities that can play an essential role in drug delivery systems. In this study, palbociclib is loaded onto the magnetic PAMAM dendrimer. Hanging droplet method was used in order to form 3D spheroids. The possible toxic effects of both free drug and drug loaded nanoparticles were evaluated in 2D and 3D MCF-7, MD-MB-231 and SKBR-3 breast cancer cell culture models by performing MTT cell viability and Alamar Blue assays. MTT analysis was performed with six different doses from 1000 µg/ml to 25 µg/ml. Drug unloaded PAMAM dendrimer did not demonstrate significant toxicity on all breast cancer cell lines. The results showed that 3D spheroids are clearly less sensitive than 2D cell cultures to free palbociclib. Also, palbociclib loaded PAMAM dendrimers showed more toxic effect than free palbociclib in all cell lines at 2D and 3D cultures. The results suggest that the traditional cell culture method (2D) is insufficient for mimicking the actual tumor tissue. The response of the cancer cells to anticancer drugs is different in the 2D and 3D culture conditions. This study showed that breast cancer cells are more resistant to free palbociclib in 3D cultures than in 2D cultures. However, nanoparticle loaded drugs can be more cytotoxic when compared to free drug.

Keywords: 2D and 3D cell culture, breast cancer, palbociclibe, PAMAM magnetic nanoparticles

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Authors: Ada Yurman

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Using the case of Israeli Atomic Spy Mordechai Vanunu as an example, this study sought to examine social response to political deviance whereby social response can be mobilized in order to achieve social control. Mordechai Vanunu, a junior technician in the Dimona Atomic Research Center, played a normative role in the militaristic discourse while working in the “holy shrine” of the Israeli defense system for many years. At a certain stage, however, Vanunu decided to detach himself from this collective and launched an assault on this top-secret circle. Israeli society in general and the security establishment in particular found this attack intolerable and unforgivable. They presented Vanunu as a ticking time bomb, delegitimized him and portrayed him as “other”. In addition, Israeli enforcement authorities imposed myriad prohibitions and sanctions on Vanunu even after his release from prison – “as will be done to he who desecrates holiness.” Social response to Vanunu at the time of his capture and trial was studied by conducting a content analysis of six contemporary daily newspapers. The analysis focused on use of language and forms of expression. In contrast with traditional content analysis methodology, this study did not just look at frequency of expressions of ideas and terms in the text and covert content; rather, the text was analyzed as a structural whole, and included examination of style, tone and unusual use of imagery, and more, in order to uncover hidden messages within the text. The social response to this case was extraordinarily intense, not only because in this case of political deviance, involving espionage and treason, Vanunu’s actions comprised a real potential threat to the country, but also because of the threat his behavior posed to the symbolic universe of society. Therefore, the response to this instance of political deviance can be seen as being part of a mechanism of social control aiming to protect world view of society as a whole, as well as to punish the criminal.

Keywords: militarism, political deviance, social construction, social control

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Authors: Lubna Azmi, Ila Shukla, Shyam Sundar Gupta, Padam Kant, Ch. V. Rao

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Gastric ulcer is a major global health threat, and it is the leading cause of stomach cancer death worldwide. Helicobacter pylori bacteriumis the most important etiologic factor for gastric ulcer. This infection is highly pervasive in South Asian developing countries, especially in India, Nepal, Srilanka etc. due to diversification in geographic area. Pathophysiology of gastric mucosal damage associated with non-invasive bacterium has not justified in detail, but it leads to change in histopathology, immunochemistry of the gastric and duodenal reason of host. The mechanism responsible for bacteria tissue tropism and mucosal damage in stomach variance during the disease is not clearly described and understood scientifically in treatment and control of pathogenic organisms. Polyphenols are secondary metabolites of plants and are generally involved in defense against aggression by pathogens. 2-(3,4-dihydroxyphenyl)-3,5,7-trihydroxychromen-4-one and 1-hydroxy-5,7-dimethoxy-2-naphthalene-carboxaldehyde are polyphenolic compound obtained from popular Indian medicinal plants ghavpatta (ArgeriaspeciosaLinn.f) andBael (Aeglemarmelos) have long been used in traditional Ayurvedic Indian medicine for various diseases. They have promising effects on ulcer, as detailed investigation has made in our laboratory. Therefore, the aim of present study is to explore membrane –dependent morphogenesis of H. pylori and associated apoptosis-mediated cell death. Based on this we analyzed immune gene expression in stomach of experimental animals with H. pylori, using quantitative reverse transcription polymerase chain reaction(q RT-PCR). This revealed rapid induction of prostaglandin, interferon I (INF-I), interferon II (INF-II) and INF-I associated genes in the infected animal. Ultrastructural changes associated with H. pylori will be taken for advanced studies. This investigation shows that the biomarkers eradicate H. pylori bacterium caused gastric ulcer which is a major risk factor for gastric cancer.

Keywords: gastric ulcer, Helicobacter pylori, immunochemistry, polyphenols

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Authors: Hyeon Sop Baek

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This paper will briefly investigate Jeong Do-jeon’s political philosophy. Jeong Do-jeon was a Korean Confucian philosopher and politician during the turbulent 14th Century who revolted against the old order, founded Joseon Dynasty, and significantly impacted the development of Korean culture. Jeong’s vision of an ideal state involved a polity that has its roots in the people -that is, an ideal government prioritizes caring for the welfare of the people, respecting and attending to the diverse opinions and concerns of the people, and relies on the genuine, voluntary support of the people. With the neo-Confucian worldview in mind -that every human being has the equal potential to become a moral person- Jeong sought to create a world suitable for everybody to contribute to the decision-making procedure and be able to realize their potential fully. This paper will first examine his works and present a quick overview of his vision of the ideal government. Then, it will examine the Confucian virtues of ren (仁) and yi (義) and how they formulate the basis of his philosophy, and then discuss the central features of his vision of government: popular mandate, equity of wealth, promoting freedom of expression and political participation, and elevating caring disposition as the paramount quality of the political leaders. Furthermore, this paper aims to analyze the element of care inherent within his political philosophy, namely his view on the dynamics of power, nurturing the people, and noncoercive justice. Finally, a discussion on why his philosophy is still relevant in the contemporary context will be provided. Jeong’s view aimed at building a sustainable model of government, by proposing that the people should be the foundation of a state and that they need to be carefully nurtured so they can realize their inborn potential and continue to contribute to the sustenance of the world, is the focal point of Jeong’s philosophy. Just as he sought to rebuild his world following the turmoils of the 14th Century, his philosophy still has a substantial implication on how we should strive to rebuild our society today.

Keywords: Korea, Confucianism, Jeong Do-jeon, Joseon, Korean philosophy, political philosophy

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Authors: Shanakr Bhattarai, V. S. Tiwari, Barak Akabayov

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The plummeting number of infections and death is due to the development of drug-resistant bacteria. In addition, the number of approved antibiotic drugs by the Food and Drug Administration (FDA) is insufficient. Therefore, developing new drugs and finding novel targets for central metabolic pathways in bacteria is urgently needed. One of the promising targets is DNA replication machinery which consists of many essential proteins and enzymes. DnaG primase is an essential enzyme and a central part of the DNA replication machinery. DnaG primase synthesizes short RNA primers that initiate the Okazaki fragments by the lagging strand DNA polymerase. Therefore, it is reasonable to assume that inhibition of primase activity will stall DNA replication and prevent bacterial proliferation. We did the expression and purification of eight different bacterial DnaGs (Mycobacterium tuberculosis(Mtb), Bacillus anthracis (Ba), Mycobacterium smegmatis (Msmeg), Francisella tularencis (Ft), Vibrio cholerae (Vc) and Yersinia pestis (Yp), Staphylococcus aureus(Saureus), Escherichia coli(Ecoli)) followed by the radioactive activity assay. After obtaining the pure and active protein DnaG, we synthesized the inhibitors for them. The inhibitors were divided into five different groups, each containing five molecules, and the cocktail inhibition assay was performed against each DnaGs. The groups of molecules inhibiting the DnaGs were further tested with individual molecules belonging to inhibiting groups. Each molecule showing inhibition was titrated against the corresponding DnaGs to find IC50. We got a molecule(VS167) that acted as broad inhibitors, inhibiting all eight DnaGs. Molecules VS180 and VS186 inhibited seven DnaGs (except Saureus). Similarly, two molecules(VS 173, VS176) inhibited five DnaGs (Mtb, Ba, Ft, Yp, Ecoli). VS261 inhibited four DnaGs (Mtb, Ba, Ft, Vc). MS50 inhibited Ba and Vc DnaGs. And some of the inhibitors inhibited only one DnaGs. Thus we found the broad and specific inhibitors for different bacterial DnaGs, and their Structure-activity analysis(SAR) was done. Further, We tried to explain the similarities among the enzyme DnaGs from different bacteria based on their inhibition pattern.

Keywords: DNA replication, DnaG, okazaki fragments, antibiotic drugs

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Authors: Eli Buchbinder

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Introduction. Language is the basis to our experience as human being. We use language in describing our experiences and construct meaning and narratives from experiences. Metaphors are a valuable linguistic tool commonly use. Metaphors link two domains that are ordinarily not related. Metaphors achieve simultaneously multi-level integration: abstract and concrete, rational and imaginative, familiar and the unfamiliar, conscious and preconscious/unconscious. As such, metaphors epistemological and ontological tool that are important in social work in every field and domain. Goals and Methods The presentation’s aim is to validate the value of metaphors through the first psychiatric breakdown is a traumatic for families. The presentation is based on two pooled qualitative studies. The first study focused on 12 spouses: 7 women and 5 men, between the ages of 22 and 57, regarding their experiences and meanings of the first psychiatric hospitalization of their partners diagnosed with affective disorders. The second study focused on 10 parents, between the ages of 47 and 62, regarding their experiences and meanings following their child's first psychotic breakdown during young adulthood. Results Two types of major metaphors evolved from the interviews in farming the trauma of the first mental breakdown. The first mode - orientation (spatial) metaphors, reflect symbolic expression of the loss of a secure base, represented in the physical environment, e.g., describing hospitalization as "falling into an abyss." The second mode- ontological metaphors, reflect how parents and spouses present their traumatic experiences of hospitalization in terms of discrete, powerful and coherent entities, e.g., describing the first hospitalization as "swimming against the tide." The two metaphors modes reflect the embodiment of the unpredictability, being mired in distress, shock, intense pain and the experience the collapse of continuity on the life course and cuts off the experience of control. Conclusions Metaphors are important and powerful guide in assessing individuals and families’ phenomenological reality. As such, metaphors are useful for understanding and orientated therapeutic intervening, in the studies above, with the first psychiatric hospitalization experienced, as well as in others social workers’ interventions.

Keywords: first mental breakdown, metaphors, family perspective, qualitative research

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Authors: Yun-Chin Chung, Nileema Divate, Gen-Hung Chen, Pei-Ru Huang, Rupesh Divate

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Many environmental factors, such as glucose concentration, ethanol, temperature, osmotic pressure and pH, decrease the production rate of ethanol using yeast as a starter. Fermentation starters with high tolerance to various stresses are always demanded for brewing industry. Trehalose, a storage carbohydrate in cell wall of yeast, plays an important role in tolerance of environmental stress by preserving integrity of plasma membrane and stabilizing proteins. Furan aldehydes are toxic to yeast and the growth rate of yeast is significantly reduced if furan aldehydes were present in the fermentation medium. In yeast, aldehyde reductase is involved in the detoxification of reactive aldehydes and consequently the growth of yeast is improved. The aims of this study were to construct a genetic recombinant Saccharomyces cerevisiae or Pichia pastoris with furfural and HMF degrading and high ethanol tolerance capacities. Yeast strains were engineered by genetic recombination for overexpression of trehalose-6-phosphate synthase gene (tps1) and aldehyde reductase gene (ari1). TPS1 gene was cloned from S. cerevisiae by reverse transcription-polymerase chain reaction (RT-PCR) and then ligated with pGAPZαC vector. The constructed vector, pGAPZC-tps1, was transformed to recombinant yeasts strain with overexpression of ari1. The transformants with pGAPZC-tps1-ari1 were generated called STA (S. cerevisiae) and PTA (P. pastoris) with overexpression of tps1, ari1. PCR with tps1-specific primers and western blot with his-tag confirmed the gene insertion and protein expression of tps1 in the transformants, respectively. The neutral trehalase gene (nth1) of STA was successfully deleted and the novel strain STAΔN will be used for further study, including the measurement of trehalose concentration and ethanol, furfural tolerance assay.

Keywords: genetic recombinant, yeast, ethanol tolerance, trehalase, aldehyde reductase

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Authors: Yi Sun, George Ed Rainger, Steve P. Watson

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Background: Beyond the classical roles in haemostasis and thrombosis, platelets are important in the initiation and development of various thrombo-inflammatory diseases. In atherosclerosis and deep vein thrombosis, for example, platelets bridge monocytes with endothelium and form heterotypic aggregates with monocytes in the circulation. This can alter monocyte phenotype by inducing their activation, stimulating adhesion and migration. These interactions involve cell surface receptor-ligand pairs on both cells. This list is likely incomplete as new interactions of importance to platelet biology are continuing to be discovered as illustrated by our discovery of PEAR-1 binding to FcεR1α. Results: We have developed a highly sensitive avidity-based assay to identify novel extracellular interactions among 126 recombinantly-expressed platelet cell surface and secreted proteins involved in platelet aggregation. In this study, we will use this method to identify novel platelet-monocyte interactions. We aim to identify ligands for orphan receptors and novel partners of well-known proteins. Identified interactions will be studied in preliminary functional assays to demonstrate relevance to the inflammatory processes supporting atherogenesis. Conclusions: Platelet-monocyte interactions are essential for the development of thromboinflammatory disease. Up until relatively recently, technologies only allow us to limit our studies on each individual protein interaction at a single time. These studies propose for the first time to study the cell surface platelet-monocyte interactions in a systematic large-scale approach using a reliable screening method we have developed. If successful, this will likely to identify previously unknown ligands for important receptors that will be investigated in details and also provide a list of novel interactions for the field. This should stimulate studies on developing alternative therapeutic strategies to treat vascular inflammatory disorders such as atherosclerosis, DVT and sepsis and other clinically important inflammatory conditions.

Keywords: membrane proteins, large-scale screening, platelets, recombinant expression

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Authors: Jorn Fricke, Frans Melissen

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Citizens in urban areas are prone to increased levels of stress due to urbanization, inadequate and overburdened infrastructure and services, and environmental degradation. Moreover, communities are fragile and subject to shocks and stresses through various social and political processes. A loss of (a sense of) community is often seen as related to increasing political and civic disintegration. Feelings of community can manifest themselves in various ways but underlying all these manifestations is the need for trust between people. One of the main drivers of trust between individuals is (shared) experiences. It is these shared experiences that may play an important role in building resilience, i.e., the ability of a community and its members to adapt to and deal with stresses, as well as ensure the ongoing development of a community. So far, experience design, as a discipline and academic field, has mainly focused on designing products or services. However, people-to-people experiences are the ones that play a pivotal role in building inclusiveness, safety, and resilience in communities. These experiences represent challenging objects of design as they develop in an interactive space of spontaneity, serendipity, and uniqueness that is based on intuition, freedom of expression, and interaction. Therefore, there is a need for research to identify which elements are required in designing the social and physical environment (or ‘experience scape’) to increase the chance for people-to-people experiences to be successful and what elements are required for these experiences to help in building resilience in urban communities that can resist shocks and stresses. By means of a comparative case study approach in urban areas in Germany and the Netherlands, using a range of qualitative research methods such as in-depth interviews, focus groups, participant observation, storytelling techniques, and life stories, this research identifies relevant actors and their roles in creating building blocks of optimal experience scrapes for building resilience in communities.

Keywords: community development, experiences, experience scapes, resilience

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Authors: Reihane Mehrparvar

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Human age could exceed further by altering gene expression through food intaking, although as a consequence of recent century eating patterns, human life-span getting shorter by emerging irregulating in autophagy mechanism, insulin, leptin, gut microbiota which are important etiological factors of type-2 diabetes, obesity, infertility, cancer, metabolic and autoimmune diseases. However, restricted calorie intake and vigorous exercise might be beneficial for losing weight and metabolic regulation in a short period but could not be implementable in the long term as a way of life. Therefore, the lack of a dietary program that is compatible with the genes of the body is essential. Sweet and high-glycemic-index (HGI) foods were associated with type-2 diabetes and cancer morbidity. The neuropsychological perspective characterizes the inclination of sweet and HGI-food consumption as addictive behavior; hence this process engages preference of gut microbiota, neural node, and dopaminergic functions. Moreover, meal composition is not the only factor that affects body hemostasis. In this narrative review, it is believed to attempt to investigate how the body responded to different food intakes and represent an accurate model based on current evidence. Eating frequently and ingesting unassimilable protein and carbohydrates may not be compatible with human genes and could cause impairments in the self-renovation mechanism. This trajectory indicates our body is more adapted to starvation and eating animal meat and marrow. Here has been recommended a model that takes into account three important factors: frequent eating, meal composition, and circadian rhythm, which may offer a promising intervention for obesity, inflammation, cardiovascular, autoimmune disorder, type-2 diabetes, insulin resistance, infertility, and cancer through intensifying autophagy-mechanism and eliminate medical costs.

Keywords: metabolic disease, anti-aging, type-2 diabetes, autophagy

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Authors: O. Benabdeljelil, A. Karioun, S. Amami, R. Rouger, M. Hamidine

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A model for preventing the risks related to climate conditions in the agricultural sector is presented. It will determine the yearly optimum premium to be paid by a producer in order to reach his required turnover. The model is based on both climatic stability and 'soft' responses of usually grown species to average climate variations at the same place and inside a safety ball which can be determined from past meteorological data. This allows the use of linear regression expression for dependence of production result in terms of driving meteorological parameters, the main ones of which are daily average sunlight, rainfall and temperature. By simple best parameter fit from the expert table drawn with professionals, optimal representation of yearly production is determined from records of previous years, and yearly payback is evaluated from minimum yearly produced turnover. The model also requires accurate pricing of commodity at N+1. Therefore, a pricing model is developed using 3 state variables, namely the spot price, the difference between the mean-term and the long-term forward price, and the long-term structure of the model. The use of historical data enables to calibrate the parameters of state variables, and allows the pricing of commodity. Application to beet sugar underlines pricer precision. Indeed, the percentage of accuracy between computed result and real world is 99,5%. Optimal premium is then deduced and gives the producer a useful bound for negotiating an offer by insurance companies to effectively protect its harvest. The application to beet production in French Oise department illustrates the reliability of present model with as low as 6% difference between predicted and real data. The model can be adapted to almost any agricultural field by changing state parameters and calibrating their associated coefficients.

Keywords: agriculture, production model, optimal price, meteorological factors, 3-factor model, parameter calibration, forward price

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Authors: Yong-Kian Lim, Khan Cheung, Xin Dang, Steven Roberts, Xiaotong Wang, Vengatesen Thiyagarajan

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Unprecedented rate of increased CO₂ level in the ocean and the subsequent changes in carbonate system including decreased pH, known as ocean acidification (OA), is predicted to disrupt not only the calcification process but also several other physiological and developmental processes in a variety of marine organisms, including edible oysters. Nonetheless, not all species are vulnerable to those OA threats, e.g., some species may be able to cope with OA stress using environmentally induced modifications on gene and protein expressions. For example, external environmental stressors, including OA, can influence the addition and removal of methyl groups through epigenetic modification (e.g., DNA methylation) process to turn gene expression “on or off” as part of a rapid adaptive mechanism to cope with OA. In this study, the above hypothesis was tested through testing the effect of OA, using decreased pH 7.4 as a proxy, on the DNA methylation pattern of an endemic and a commercially important estuary oyster species, Crassostrea hongkongensis, at the time of larval habitat selection and metamorphosis. Larval growth rate did not differ between control pH 8.1 and treatment pH 7.4. The metamorphosis rate of the pediveliger larvae was higher at pH 7.4 than those in control pH 8.1; however, over one-third of the larvae raised at pH 7.4 failed to attach to an optimal substrate as defined by biofilm presence. During larval development, a total of 130 genes were differentially methylated across the two treatments. The differential methylation in the larval genes may have partially accounted for the higher metamorphosis success rate under decreased pH 7.4 but with poor substratum selection ability. Differentially methylated loci were concentrated in the exon regions and appear to be associated with cytoskeletal and signal transduction, oxidative stress, metabolic processes, and larval metamorphosis, which implies the high potential of C. hongkongensis larvae to acclimate and adapt through non-genetic ways to OA threats within a single generation.

Keywords: adaptive plasticity, DNA methylation, larval metamorphosis, ocean acidification

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Authors: Mehrdad Abkenari, Orod Zarrinkafsh, Mohsen Ramezan Shirazi

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Construction projects initiate in complicated dynamic environments and, due to the close relationships between project parameters and the unknown outer environment, they are faced with several uncertainties and risks. Success in time, cost and quality in large scale construction projects is uncertain in consequence of technological constraints, large number of stakeholders, too much time required, great capital requirements and poor definition of the extent and scope of the project. Projects that are faced with such environments and uncertainties can be well managed through utilization of the concept of risk management in project’s life cycle. Although the concept of risk is dependent on the opinion and idea of management, it suggests the risks of not achieving the project objectives as well. Furthermore, project’s risk analysis discusses the risks of development of inappropriate reactions. Since evaluation and prioritization of construction projects has been a difficult task, the network structure is considered to be an appropriate approach to analyze complex systems; therefore, we have used this structure for analyzing and modeling the issue. On the other hand, we face inadequacy of data in deterministic circumstances, and additionally the expert’s opinions are usually mathematically vague and are introduced in the form of linguistic variables instead of numerical expression. Owing to the fact that fuzzy logic is used for expressing the vagueness and uncertainty, formulation of expert’s opinion in the form of fuzzy numbers can be an appropriate approach. In other words, the evaluation and prioritization of construction projects on the basis of risk factors in real world is a complicated issue with lots of ambiguous qualitative characteristics. In this study, evaluated and prioritization the risk parameters and factors with fuzzy logy method by combination of three method DEMATEL (Decision Making Trial and Evaluation), ANP (Analytic Network Process) and TOPSIS (Technique for Order-Preference by Similarity Ideal Solution) on Construction Management.

Keywords: fuzzy logy, risk, prioritization, assessment

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Authors: Neeraj Shetye

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A common phenomenon between the rise of leaders like Mussolini and Adolf Hitler in the 20th century is their 'charisma'. They possessed the ability to seduce the crowd not just by the things they said but also by the way they said it. Aristotle defined rhetoric as an art of persuasion and reasoning which is how social scientists understand the concept. Political rhetoric in a modern democracy has several complexities including the huge number of speakers, quantity of information, diverse viewpoints, number of candidates and the impact of digital age. Politics in India since the seventies have been 'visibly dominated' by uses of rhetorical language and with a number of slogans. This idea of how language can steer an individual to establish or adopt a certain viewpoint has not been a focus of study in the Indian discourse. In a linguistically diverse region such as India, the idea of political rhetoric is vast and may not be accomplished in a year. There are in-depth studies by western thinkers on European or American political rhetoric unlike their Asian counterparts such as China, India or any of the Pacific nations. India saw an integration of states based on languages. Keeping this idea in mind, this paper aims to cover one political party that rose to its prominence over five decades and most significantly known for its conservative expression: Shivsena. Shivsena’s rise during the eighties and eventually establishing their government in the nineties are two fascinating periods to focus especially with a simultaneous rise of Bombay underworld, Babri demolition and major economic policy changes in the form of liberalisation (1991) and globalization (1995). This project attempts to study this with a two-fold methodology: literature review and fieldwork. There is an immense literature on Shivsena by both its admirers and critiques, contributing to both sides of the debate. Scholars have been writing about this party over these years and have keenly observed its growing popularity amongst the masses. There is just one intention behind this project, and it is to connect and analyse the vast, dispersed literature that is available and contribute to a field that has not been adequately analysed in the academic discourse.

Keywords: India, language, political rhetoric, Shivsena, slogans

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Authors: Qin Yang, Fan Zhang, Juan Du, Chongkui Sun, Krishna Kota, Yun-Ling Zheng

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Telomeres are specialized structures at chromosome ends consisting of tandem repetitive DNA sequences [(TTAGGG)n in humans] and associated proteins, which are necessary for telomere function. Telomere lengths are tightly regulated within a narrow range in normal human somatic cells, the basis of cellular senescence and aging. Previous studies have extensively focused on how short telomeres are extended and have demonstrated that telomerase plays a central role in telomere maintenance through elongating the short telomeres. However, the molecular mechanisms of regulating excessively long telomeres are unknown. Here, we found that telomerase enzymatic component hTERT plays a dual role in the regulation of telomeres length. We analyzed single telomere alterations at each chromosomal end led to the discoveries that hTERT shortens excessively long telomeres and elongates short telomeres simultaneously, thus maintaining the optimal telomere length at each chromosomal end for an efficient protection. The hTERT-mediated telomere shortening removes large segments of telomere DNA rapidly without inducing telomere dysfunction foci or affecting cell proliferation, thus it is mechanistically distinct from rapid telomere deletion. We found that expression of hTERT generates telomeric circular DNA, suggesting that telomere homologous recombination may be involved in this telomere shortening process. Moreover, the hTERT-mediated telomere shortening is required its enzymatic activity, but telomerase RNA component hTR is not involved in it. Furthermore, shelterin protein TPP1 interacts with hTERT and recruits it on telomeres to mediate telomere shortening. In addition, telomere-associated proteins, DKC1 and TCAB1 also play roles in this process. This novel hTERT-mediated telomere shortening mechanism not only exists in cancer cells, but also in primary human cells. Thus, the hTERT-mediated telomere shortening is expected to shift the paradigm on current molecular models of telomere length maintenance, with wide-reaching consequences in cancer and aging fields.

Keywords: aging, hTERT, telomerase, telomeres, human cells

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Authors: Saima Murtaza

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Ethics and aesthetics have invariably remained the two closely integrated artistic appurtenances for the production of any work of art. These artistic devices configure themselves into a complex synthesis in our contemporary literature. The labyrinthine integration of ethics and aesthetics, operating in the lives of human characters, to the extent of transcending all limits has resulted in an artistic puzzle for the readers. Art, no doubt, is an extrinsic expression of the intrinsic life of man. The use of aesthetics in literature pertaining to human existence; aesthetic solipsism, has resulted in the artistic objectification of these characters. The practice of the like aestheticism deprives the characters of their souls, rendering them as mere objects of aesthetic gaze at the hands of their artists-creators. Artists orchestrate their lives founding it on a plot which deviates from normal social and ethical standards. Their perverse attitude can be seen in dealing with characters, their feelings and the incidents of their lives. Morality is made to appear not as a religious construct but as an individual’s private affair. Furthermore, the idea of beauty incarnated, in other words hedonistic aesthetic does not placate a true aesthete. Ethics and aesthetics are the two most recurring motifs of our contemporary literature, especially of Nabokov’s world. The purpose of this study is to peruse these aforementioned motifs in Nabokov’s most enigmatic novel Lolita, a story of pedophilia, which is in fact reflective of our complex individual psychic and societal patterns. The narrative subverts all the traditional and hitherto known notions of aesthetics and ethics. When applied to literature, aesthetic does not simply mean ‘beautiful’ in the text. It refers to an intricate relationship between feelings and perception and also incorporates within its range wide-ranging emotional reactions to text. The term aesthetics in literature is connected with the readers whose critical responses to the text determine the merit of any work to be really a piece of art. Aestheticism is the child of ethics. Morality sets the grounds for the production of any work and the idea of aesthetics gives it transcendence.

Keywords: ethics, aesthetics and hedonistic aesthetic, nymphet syndrome, pedophilia

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Authors: Pei Chi Fang, Wei Chen Lin

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Trichomonas tenax is a microaerophilic oral protozoan found in patients with poor oral hygiene. It participates in the inflammatory process of periodontal disease and can potentially be aspirated into the lungs, giving rise to pulmonary trichomoniasis. However, the precise roles of T. tenax in the pulmonary system remain largely unexplored and warrant comprehensive epidemiological investigation. To assess the prevalence of T. tenax infection, we collected bronchoalveolar lavage fluid (BALF) samples from hospitalized patients with lung diseases. A specific nested PCR approach was employed to determine prevalence rates, yielding 21 positive cases out of 61 samples from Ditmanson Medical Foundation Chia-Yi Christian Hospital, and 11 positive cases out of 55 samples from National Cheng Kung University Hospital. Furthermore, there is a critical need for comprehensive data regarding the presence of T. tenax in environmental surface watersheds. In this context, we present findings from investigations in the Yanshuei and Donggang river basins in southern Taiwan, which are crucial sources for public drinking water in the region. In order to elucidate potential implications on pulmonary virus infections, we conducted an analysis of gene expression level changes in H292 cell line after exposure to T. tenax. Our findings revealed significant regulation of multiple virus-related genes, including IFI44L and IFITM3. Ongoing research endeavors are focused on identifying the key components within T. tenax responsible for these observed effects. Crucially, this study lays the groundwork for a preliminary understanding of T. tenax prevalence in patients with pulmonary diseases. It also seeks to establish a meaningful correlation between lung infections and oral hygiene practices, with the ultimate aim of informing distinct treatment and prevention strategies.

Keywords: parasitology, genes, virus, human health, infection, lung

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Authors: Hanan Khojah, RuAngelie Edrada-Ebel

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Neurodegenerative disease including Alzheimer’s disease is a major cause of long-term disability. Oxidative stress is frequently implicated as one of the key contributing factors to neurodegenerative diseases. Protection against neuronal damage remains a great challenge for researchers. Ficus carica (commonly known as fig) is a species of great antioxidant nutritional value comprising a protective mechanism against innumerable health disorders related to oxidative stress as well as Alzheimer’s disease. The purpose of this work was to characterize the non-polar active metabolites in Ficus carica endocarp, mesocarp, and exocarp. Crude extracts were prepared using several extraction solvents, which included 1:1 water: ethylacetate, acetone and methanol. The dried extracts were then solvent partitioned between equivalent amounts of water and ethylacetate. Purification and fractionation were accomplished by high-throughput chromatography. The isolated metabolites were tested on their effect on human neuroblastoma cell line by cell viability test and cell cytotoxicity assay with acrolein. Molecular weights of the active metabolites were determined via LC–HRESIMS and GC-EIMS. Metabolomic profiling was performed to identify the active metabolites by using differential expression analysis software (Mzmine) and SIMCA for multivariate analysis. Structural elucidation and identification of the interested active metabolites were studied by 1-D and 2-D NMR. Significant differences in bioactivity against a concentration-dependent assay on acrolein radicals were observed between the three fruit parts. However, metabolites obtained from mesocarp and the endocarp demonstrated bioactivity to scavenge ROS radical. NMR profiling demonstrated that aliphatic compounds such as γ-sitosterol tend to induce neuronal bioactivity and exhibited bioactivity on the cell viability assay. γ-Sitosterol was found in higher concentrations in the mesocarp and was considered as one of the major phytosterol in Ficus carica.

Keywords: alzheimer, Ficus carica, γ-Sitosterol, metabolomics

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Authors: M. Uysal, M. Yilmaz, I. Tiryakioğlu

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Digital Terrain Model (DTM) is a digital numerical representation of the Earth's surface. DTMs have been applied to a diverse field of tasks, such as urban planning, military, glacier mapping, disaster management. In the expression of the Earth' surface as a mathematical model, an infinite number of point measurements are needed. Because of the impossibility of this case, the points at regular intervals are measured to characterize the Earth's surface and DTM of the Earth is generated. Hitherto, the classical measurement techniques and photogrammetry method have widespread use in the construction of DTM. At present, RADAR, LiDAR, and stereo satellite images are also used for the construction of DTM. In recent years, especially because of its superiorities, Airborne Light Detection and Ranging (LiDAR) has an increased use in DTM applications. A 3D point cloud is created with LiDAR technology by obtaining numerous point data. However recently, by the development in image mapping methods, the use of unmanned aerial vehicles (UAV) for photogrammetric data acquisition has increased DTM generation from image-based point cloud. The accuracy of the DTM depends on various factors such as data collection method, the distribution of elevation points, the point density, properties of the surface and interpolation methods. In this study, the random data reduction method is compared for DTMs generated from image based point cloud data. The original image based point cloud data set (100%) is reduced to a series of subsets by using random algorithm, representing the 75, 50, 25 and 5% of the original image based point cloud data set. Over the ANS campus of Afyon Kocatepe University as the test area, DTM constructed from the original image based point cloud data set is compared with DTMs interpolated from reduced data sets by Kriging interpolation method. The results show that the random data reduction method can be used to reduce the image based point cloud datasets to 50% density level while still maintaining the quality of DTM.

Keywords: DTM, Unmanned Aerial Vehicle (UAV), uniform, random, kriging

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Authors: Md. Alauddin, Md. Ruhul Amin, Md. Omar Faruque, Muhammad Ali Siddiquee, Zakir Hossain Howlader, Mohammad Asaduzzaman

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Diabetes and hypertension are the major lifestyle related diseases. The α-amylase and angiotensin converting enzymes (ACE) are the key enzymes that regulate diabetes and hypertension. The aim was to develop a drug for the treatment of diabetes and hypertension. The Rice Bran (RB) sample (Oryza sativa; BRRI-Dhan-84) was collected from the Bangladesh Rice Research Institute (BRRI), and rice bran proteins were isolated and hydrolyzed by hydrolyzing enzyme alcalase and trypsin. In vivo experiment suggested that rice bran bioactives has an effect on regulating the expression of several key gluconeogenesis and lipogenesis-regulating genes, such as glucose-6-phosphatase, phosphoenolpyruvate carboxykinase, and fatty acid synthase. The above genes have a connection of regulating the glucose level, lipids profile as well as act as an anti-inflammatory agent. A molecular docking, bioinformatics and in vitro experiments were performed. We found rice bran protein hydrolysates significantly (<0.05) influence the peptide concentration in the case of trypsin, alcalase, and (trypsin + alcalase) digestion. The in vitro analysis found that protein hydrolysate significantly (<0.05) reduced diabetic and hypertension as well as oxidative stress. A molecular docking study showed that the YY and IP peptide have a significantly strong binding affinity to the active site of the ACE enzyme and α-amylase with -7.8Kcal/mol and -6.2Kcal/mol, respectively. The Molecular dynamics (MD) simulation and Swiss ADME data analysis showed that less toxicity risk, good physicochemical properties, pharmacokinetics, and drug-likeness with drug scores 0.45 and 0.55 of YY and IP peptides, respectively. Thus, rice bran bioactive could be a good candidate for the treatment of diabetes and hypertension.

Keywords: anti-hypertensive and anti-hyperglycemic, anti-oxidative, bioinformatics, in vitro study, rice bran proteins and peptides

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Authors: Madhuri Goswami

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The matrix of power establishes the hegemonic dominance and supremacy of one group through exercising repression and relegation upon the other. However, the injustice done to any race, ethnicity, or caste has instigated the protest and resistance through various modes -social campaigns, political movements, literary expression and so on. Consequently, the search for identity, the means of claiming it and strive for recognition have evolved as the persistent phenomena all through the world. In the discourse of protest and minority literature, these two discourses -African American and Indian Dalit- surprisingly, share wrath and anger, hope and aspiration, and quest for identity and struggle for self-assertion. African American and Indian Dalit are two geographically and culturally apart communities that stand together on a single platform. This paper has sought to comprehend the form and investigate the formation of identity in general and in the literary work of Toni Morrison and Indian Dalit writing, particular, i.e., Black identity and Dalit identity. The study has speculated two types of identity, namely, individual or self and social or collective identity in the literary province of these marginalized literature. Morrison’s work outsources that self-identity is not merely a reflection of an inner essence; it is constructed through social circumstances and relations. Likewise, Dalit writings too have a fair record of discovery of self-hood and formation of identity, which connects to the realization of self-assertion and worthiness of their culture among Dalit writers. Bama, Pawar, Limbale, Pawde, and Kamble investigate their true self concealed amid societal alienation. The study has found that the struggle for recognition is, in fact, the striving to become the definer, instead of just being defined; and, this striving eventually, leads to the introspection among them. To conclude, Morrison as well as Indian marginalized authors, despite being set quite distant, communicate the relation between individual and community in the context of self-consciousness, self-identification and (self) introspection. This research opens a scope for further research to find out similar phenomena and trace an analogy in other world literatures.

Keywords: identity, introspection, self-access, struggle for recognition

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Authors: Axel Mancebo, Ana M. Bada, Angel Casacó, Bárbara González, Avelina León, María E. Arteaga, Consuelo González, Belinda Sánchez, Adriana Carr, Nuris Ledón, Arianna Iglesias

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Despite the many preventive and therapeutic modalities aimed at curing cancer, it remains as a serious world health problem. Promising recent developments suggest that cancer immunotherapy may be the next great hope for cancer treatment. EGFRs are receptor tyrosine kinases and it is considered an important therapeutic target related with tumor progression, and several types of molecular therapies, including monoclonal antibodies, small molecules, and vaccines, have been developed to target the HER family of receptors. On the other hand, gangliosides are membrane glycosphingolipids that contain two variants of sialic acid, the N-acetylated (NeuAc) and the N-glycolylated (NeuGc) variant. The high expression of this antigen-specific molecule has been associated with malignant tumor progression and immunosuppressive mechanisms, so ganglioside could be considered as the target for cancer immunotherapy. We have been working for several years in the safety evaluation of cancer vaccines targeting these two systems, the EGF receptor and ganglioside. We presented in this work results of repeated dose toxicity studies performed in Sprague Dawley rats and Cynomolgus monkeys, including clinical observations, body weight and rectal temperature measuring, clinical pathology analysis, gross necropsy and histological examination in rodent studies, and immunological evaluation. Immunizations were capable of inducing mainly inflammatory effects at the injection site, with findings largely attributable to the adjuvants used and probably enhanced by the immunological properties of the antigens. In general, these vaccines were shown to be well tolerated, and these studies in relevant species allow treating cancer patients with tumors during long periods with relative weight safety margin.

Keywords: cancer vaccines, safety, toxicology, rats, non human primates

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Authors: Neuma Chaveiro, Juliana Guimarães Faria

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Introduction: The thematic that guides this study is teacher training for the teaching of sign language in a perspective of bilingual education – specifically aimed at Brazilian public schools that offer inclusive education, and that have, among its students, deaf children who use Brazilian Sign Language as a means of communication and expression. In the Teacher Training Course for Letters/Libras at the Universidade Federal de Goiás/UFG, we developed a bilingual education project for the deaf, linked to PIBID (Institutional Scholarship for Teaching Initiation Program), funded by the Brazilian Federal Government through CAPES (Coordination for the Improvement of Higher Education Personnel). Goals: to provide the education of higher education teachers to work in public schools in basic education and to insert students from the UFG’s Letters/Libras course in the school’s daily life, giving them the opportunity for the creation and participation in methodological experiences and of teaching practices in order to overcome the problems identified in the teaching-learning process of deaf students, in a bilingual perspective, associating Libras (Brazilian Sign Language) and Portuguese. Methodology: qualitative approach and research-action, prioritizing action – reflection – action of the people involved. The Letters-Libras PIBID of the College of Letters/UFG, in this qualitative context, is guided by the assumptions of investigation-action to contribute to the education of the Libras teacher. Results: production of studies and researches in the area of education, professionalization and teaching practice for the degree holder in Letters: Libras; b) studies, research and training in bilingual education; c) clarification and discussion of the myths that permeate the reality of users of sign languages; d) involving students in the development of didactic materials for bilingual education. Conclusion: the PIBID Project Letters/Libras allows, both to the basic education school and to the teachers in training for the teaching of Libras, an integrated and collective work partnership, with discussions and changes in relation to bilingual education for the deaf and the teaching of Libras.

Keywords: deaf, sign language, teacher training, educacion

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Authors: Ravi Ande, Mousumi Hazari

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One of the common geophysical techniques for mapping subsurface geo-electrical structures, extensive hydro-geological research, and engineering and environmental geophysics applications is the use of time domain electromagnetic (TDEM)/transient electromagnetic (TEM) soundings. A large transmitter loop for energising the ground and a small receiver loop or magnetometer for recording the transient voltage or magnetic field in the air or on the surface of the earth, with the receiver at the center of the loop or at any random point inside or outside the source loop, make up a large loop TEM system. In general, one can acquire data using one of the configurations with a large loop source, namely, with the receiver at the center point of the loop (central loop method), at an arbitrary in-loop point (in-loop method), coincident with the transmitter loop (coincidence-loop method), and at an arbitrary offset loop point (offset-loop method), respectively. Because of the mathematical simplicity associated with the expressions of EM fields, as compared to the in-loop and offset-loop systems, the central loop system (for ground surveys) and coincident loop system (for ground as well as airborne surveys) have been developed and used extensively for the exploration of mineral and geothermal resources, for mapping contaminated groundwater caused by hazardous waste and thickness of permafrost layer. Because a proper analytical expression for the TEM response over the layered earth model for the large loop TEM system does not exist, the forward problem used in this inversion scheme is first formulated in the frequency domain and then it is transformed in the time domain using Fourier cosine or sine transforms. Using the EMLCLLER algorithm, the forward computation is initially carried out in the frequency domain. As a result, the EMLCLLER modified the forward calculation scheme in NLSTCI to compute frequency domain answers before converting them to the time domain using Fourier Cosine and/or Sine transforms.

Keywords: time domain electromagnetic (TDEM), TEM system, geoelectrical sounding structure, Fourier cosine

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Authors: Neha Devi, P. K. Joshi

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Fire fuel map is one of the most critical factors for planning and managing the fire hazard and risk. One of the most significant forms of global disturbance, impacting community dynamics, biogeochemical cycles and local and regional climate across a wide range of ecosystems ranging from boreal forests to tropical rainforest is wildfire Assessment of fire danger is a function of forest type, fuelwood stock volume, moisture content, degree of senescence and fire management strategy adopted in the ground. Remote sensing has potential of reduction the uncertainty in mapping fuels. Hyperspectral remote sensing is emerging to be a very promising technology for wildfire fuels characterization. Fine spectral information also facilitates mapping of biophysical and chemical information that is directly related to the quality of forest fire fuels including above ground live biomass, canopy moisture, etc. We used Hyperion imagery acquired in February, 2016 and analysed four fuel characteristics using Hyperion sensor data on-board EO-1 satellite, acquired over the Shiwalik Himalayas covering the area of Champawat, Uttarakhand state. The main objective of this study was to present an overview of methodologies for mapping fuel properties using hyperspectral remote sensing data. Fuel characteristics analysed include fuel biomass, fuel moisture, and fuel condition and fuel type. Fuel moisture and fuel biomass were assessed through the expression of the liquid water bands. Fuel condition and type was assessed using green vegetation, non-photosynthetic vegetation and soil as Endmember for spectral mixture analysis. Linear Spectral Unmixing, a partial spectral unmixing algorithm, was used to identify the spectral abundance of green vegetation, non-photosynthetic vegetation and soil.

Keywords: forest fire fuel, Hyperion, hyperspectral, linear spectral unmixing, spectral mixture analysis

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Authors: Mona Alharbi

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Melioidosis has emerged as a lethal disease. Unfortunately, the molecular mechanisms of virulence and pathogenicity of Burkholderia pseudomallei remain unknown. However, proteomics research has selected putative targets in B. pseudomallei that might play roles in the B. pseudomallei virulence. BPSL 2418 putative protein has been predicted as a free methionine sulfoxide reductase and interestingly there is a link between the level of the methionine sulfoxide in pathogen tissues and its virulence. Therefore in this work, we describe the cloning expression, purification, and crystallization of BPSL 2418 and the solution of its 3D structure using X-ray crystallography. Also, we aimed to identify the substrate binding and reduced forms of the enzyme to understand the role of BPSL 2418. The gene encoding BPSL2418 from B. pseudomallei was amplified by PCR and reclone in pETBlue-1 vector and transformed into E. coli Tuner DE3 pLacI. BPSL2418 was overexpressed using E. coli Tuner DE3 pLacI and induced by 300μM IPTG for 4h at 37°C. Then BPS2418 purified to better than 95% purity. The pure BPSL2418 was crystallized with PEG 4000 and PEG 6000 as precipitants in several conditions. Diffraction data were collected to 1.2Å resolution. The crystals belonged to space group P2 21 21 with unit-cell parameters a = 42.24Å, b = 53.48Å, c = 60.54Å, α=γ=β= 90Å. The BPSL2418 binding MES was solved by molecular replacement with the known structure 3ksf using PHASER program. The structure is composed of six antiparallel β-strands and four α-helices and two loops. BPSL2418 shows high homology with the GAF domain fRMsrs enzymes which suggest that BPSL2418 might act as methionine sulfoxide reductase. The amino acids alignment between the fRmsrs including BPSL 2418 shows that the three cysteines that thought to catalyze the reduction are fully conserved. BPSL 2418 contains the three conserved cysteines (Cys⁷⁵, Cys⁸⁵ and Cys¹⁰⁹). The active site contains the six antiparallel β-strands and two loops where the disulfide bond formed between Cys⁷⁵ and Cys¹⁰⁹. X-ray structure of free methionine sulfoxide binding and native forms of BPSL2418 were solved to increase the understanding of the BPSL2418 catalytic mechanism.

Keywords: X-Ray Crystallography, BPSL2418, Burkholderia pseudomallei, Melioidosis

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Authors: Qi LI, Xu Lin, Nengming Lin

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Objective: The aim of this study was to investigate the role of desmocollin 2 (DSC2) protein in the proliferation, migration and drug resistance of lung cancer cells. Method: CCK-8 assays and colony formation assays were used to evaluate the effect of dsc2 regulation on cancer cell viability and colony formation. Transwell assays and wound healing assays were also performed. Cell flow double staining was used to detect the apoptosis rate of cells with DSC2, which was added cisplatin. Western blot assay was used to detect cell cycle, PI3k/Akt and apoptosis-related proteins. Results: Our data showed that dsc2 is upregulated in clinical lung cancer tissues compared with pericarcinomatous tissues, and it is differentially expressed in lung cancer cell lines. The down-regulation of dsc2 in A549 and H358 lung cancer cells significantly suppressed the cell proliferation, metastasis, and motility. In contrast, the opposite effects were observed in overexpression of dsc2 both in H23 and PC9 cell lines. In addition to lung adenocarcinoma cell lines, we also examined its expression in lung squamous cell lines, such as H226. Western blotting showed that dsc2 could reduce the level of phosphorylated Akt (Ser 473) and p-mTOR. Thus, it is speculated that dsc2 up-regulation promotes proliferation and invasiveness through activation of the PI3K/AKT pathway. Also, knockdown of dsc2 in A549 and H226 could significantly decreased in the levels of cyclinB and wee1 protein. Additionally, flow cytometry showed that dsc2 knockdown combined with cisplatin could significantly enhance cell apoptosis rate. Conclusion: These data suggest that dsc2 promotes the proliferation and migration of lung cancer cells in vitro. Also, the results suggested that dsc2 could affect the cell cycle and apoptosis of lung cells. Furthermore, knockdown of dsc2 could sensitize cisplatin in both lung adenocarcinoma and lung squamous cell lines. Thus we suggested that dsc2 can be used as a therapeutic target for lung cancer.

Keywords: desmocollin 2, cisplatin, lung cancer, PI3K/AKT, lung squamous cell

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Authors: Ahn Christine Myunghee, Sim Wonjin, Cho Kristina, Ahn Mira, Hong Yeju, Kwok Jihae, Lim Sooyeon, Park Hansol

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With a sharp increase in the prevalence rate for mental health issues in Korea, there is a need for specific and effective intervention strategies in counseling and psychotherapy for use with Korean clients. With the cultural emphasis on restraining emotional expression and not disclosing personal and familial problems to outsiders, clients often find it difficult to discuss their emotional issues even to therapists. Exploring a client’s internal psychological processes bypassing this culture-specific mode of therapeutic communication often becomes a challenge in the therapeutic setting. Given this socio-cultural context, the purpose of the current study was to investigate the effectiveness of using dream work to individuals in Korea. The current study conducted one 60-90 minute dream session and analyzed the dream content of 39 Korean young adults to evaluate the effectiveness of the Hill dream model in accessing the intra-psychic materials, determining essential emotional themes, and learning how the individuals interpreted the contents of their dreams. The transcribed data, which included a total of 39 sessions from 39 volunteer university students, were analyzed by the Consensus Qualitative Research (CQR) approach in terms of domains and core ideas. Self-report measures on Dream Salience, Gains from Dream Interpretations and the Session Evaluation Scale were administered before and after each of their dream sessions. The results indicated that dream work appears to be an effective way to understand unconscious motivations, thoughts, and feelings related to a person’s sense of self, and also how these people relate to other people. Current findings need to be replicated with clients referred for counseling and psychotherapy to determine if the dream work is an appropriate and useful intervention in counseling settings. Limitations of the current study and suggestions for future follow-ups are included in the discussion.

Keywords: dream work, dream interpretation, Korean, young adults, CQR

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Authors: Snehal D. Ganjave, Hardik Dodia, Avinash V. Sunder, Swati Madhu, Pramod P. Wangikar

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Batch cultivation of recombinant bacteria in shake flasks results in low cell density due to nutrient depletion. Previous protocols on high cell density cultivation in shake flasks have relied mainly on controlled release mechanisms and extended cultivation protocols. In the present work, we report an optimized fed-batch process for high cell density cultivation of recombinant E. coli BL21(DE3) for protein production. A cybernetic model-based, multi-objective optimization strategy was implemented to obtain the optimum operating variables to achieve maximum biomass and minimized substrate feed rate. A syringe pump was used to feed a mixture of glycerol and yeast extract into the shake flask. Preliminary experiments were conducted with online monitoring of dissolved oxygen (DO) and offline measurements of biomass and glycerol to estimate the model parameters. Multi-objective optimization was performed to obtain the pareto front surface. The selected optimized recipe was tested for a range of proteins that show different extent soluble expression in E. coli. These included eYFP and LkADH, which are largely expressed in soluble fractions, CbFDH and GcanADH , which are partially soluble, and human PDGF, which forms inclusion bodies. The biomass concentrations achieved in 24 h were in the range 19.9-21.5 g/L, while the model predicted value was 19.44 g/L. The process was successfully reproduced in a standard laboratory shake flask without online monitoring of DO and pH. The optimized fed-batch process showed significant improvement in both the biomass and protein production of the tested recombinant proteins compared to batch cultivation. The proposed process will have significant implications in the routine cultivation of E. coli for various applications.

Keywords: cybernetic model, E. coli, high cell density cultivation, multi-objective optimization

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Authors: Jakub Vávra, Milan Valenta, Petr Kosek

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This article presents a perspective on the conduct of the dramatherapy process with persons with addictive behaviours with regard to the directiveness of the process. Although drama therapy as one of the creative arts approaches is rather non-directive in nature, depending on the clientele, there may be a need to structure the process more and, depending on the needs of the clients, to guide the process more directive. The specificity for people with addictive behaviours is discussed through the prism of the dramatherapeutic perspective, where we can find both a psychotherapeutic component as well as a component touching on expression and art, which is rather non-directive in nature. Within the context of practice with clients, this theme has repeatedly emerged and dramatherapists themselves have sought to find ways of coping with clients' demands and needs for structure and guidance within the dramatherapy process. Some of the outcomes from the supervision work also guided the research. Based on this insight, the research questions were approached. The first research question asks: in what ways is directive in dramatherapy manifested and manifested in the process? The second research question then complements the first and asks: to which phenomena are directivity in dramatherapy linked? In relation to the research questions, data were collected using focus groups and field notes. The qualitative approach of Content analysis and Relational analysis was chosen as the methodology. For analyzing qualitative research, we chose an Inductive coding scheme: Open coding, Axial coding, Pattern matching, Member checking, and Creating a coding scheme. In the presented partial research results, we find recurrent schemes related to directive coding in drama therapy. As an important element, directive leadership emerges in connection with safety for the client group, then in connection with the clients' order and also the department of the facility, and last but not least, to the personality of the drama therapist. By careful analysis and looking for patterns in the research results, we can see connections that are impossible to interpret at this stage but already provide clues to our understanding of the topic and open up further avenues for research in this area.

Keywords: dramatherapy, directivity, personal approach, aims of dramatherapy process, safetyness

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