Search results for: genes expression
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 2493

Search results for: genes expression

393 The Subcellular Localisation of EhRRP6 and Its Involvement in Pre-Ribosomal RNA Processing in Growth-Stressed Entamoeba histolytica

Authors: S. S. Singh, A. Bhattacharya, S. Bhattacharya

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The eukaryotic exosome complex plays a pivotal role in RNA biogenesis, maturation, surveillance and differential expression of various RNAs in response to varying environmental signals. The exosome is composed of evolutionary conserved nine core subunits and the associated exonucleases Rrp6 and Rrp44. Rrp6p is crucial for the processing of rRNAs, other non-coding RNAs, regulation of polyA tail length and termination of transcription. Rrp6p, a 3’-5’ exonuclease is required for degradation of 5’-external transcribed spacer (ETS) released from the rRNA precursors during the early steps of pre-rRNA processing. In the parasitic protist Entamoeba histolytica in response to growth stress, there occurs the accumulation of unprocessed pre-rRNA and 5’ ETS sub fragment. To understand the processes leading to this accumulation, we looked for Rrp6 and the exosome subunits in E. histolytica, by in silico approaches. Of the nine core exosomal subunits, seven had high percentage of sequence similarity with the yeast and human. The EhRrp6 homolog contained exoribonuclease and HRDC domains like yeast but its N- terminus lacked the PMC2NT domain. EhRrp6 complemented the temperature sensitive phenotype of yeast rrp6Δ cells suggesting conservation of biological activity. We showed 3’-5’ exoribonuclease activity of EhRrp6p with in vitro-synthesized appropriate RNAs substrates. Like the yeast enzyme, EhRrp6p degraded unstructured RNA, but could degrade the stem-loops slowly. Furthermore, immunolocalization revealed that EhRrp6 was nuclear-localized in normal cells but was diminished from nucleus during serum starvation, which could explain the accumulation of 5’ETS during stress. Our study shows functional conservation of EhRrp6p in E.histolytica, an early-branching eukaryote, and will help to understand the evolution of exosomal components and their regulatory function.

Keywords: entamoeba histolytica, exosome complex, rRNA processing, Rrp6

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392 Understanding Team Member Autonomy and Team Collaboration: A Qualitative Study

Authors: Ayşen Bakioğlu, Gökçen Seyra Çakır

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This study aims to explore how research assistants who work in project teams experience team member autonomy and how they reconcile team member autonomy with team collaboration. The study utilizes snowball sampling. 20 research assistants who work the faculties of education in Marmara University and Yıldız Technical University have been interviewed. The analysis of data involves a content analysis MAXQDAPlus 11 which is a qualitative data analysis software is used as the data analysis tool. According to the findings of this study, emerging themes include team norm formation, team coordination management, the role of individual tasks in team collaboration, leadership distribution. According to the findings, interviewees experience team norm formation process in terms of processes, which pertain to task fulfillment, and processes, which pertain to the regulation of team dynamics. Team norm formation process instills a sense of responsibility amongst individual team members. Apart from that, the interviewees’ responses indicate that the realization of the obligation to work in a team contributes to the team norm formation process. The participants indicate that individual expectations are taken into consideration during the coordination of the team. The supervisor of the project team also has a crucial role in maintaining team collaboration. Coordination problems arise when an individual team member does not relate his/her academic field with the research topic of the project team. The findings indicate that the leadership distribution in the project teams involves two leadership processes: leadership distribution which is based on the processes that focus on individual team members and leadership distribution which is based on the processes that focus on team interaction. Apart from that, individual tasks serve as a facilitator of collaboration amongst team members. Interviewees also indicate that individual tasks also facilitate the expression of individuality.

Keywords: project teams in higher education, research assistant teams, team collaboration, team member autonomy

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391 Paradigm Shift of the World Is Globalization: Identity Crisis, Violence and Cultural War

Authors: Shahla Bukhtair

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A paradigm presents a consensus view of a particular or collective community, accepted into by the members of that community, either consciously pronounced or, more likely, simply assumed and not intentionally acknowledged but is articulated. Paradigm shift is based on the behavioral attitude of the community. Change is inexorable. The world is suffering with the innovative creation of globalization. Media boosted this paradigm shift all over the world. Globalization is a vigorous process which impacts differentially on various cultures around the world. The outcome of the globalization is permeates cultural boundaries and in the process results in the spread of Western ideologies and values across the world. The term flourished in 20th century. Globalization is regarded as having substantial impact on such crises through its encouragement of conflicts rather than conciliation; through opportunities of expression, various groups get benefit with it. Identity crisis refers to inflexible mechanism i.e. cultural and political conflicts among polarized groups, which struggle with each other over the definition of a national identity. Violence is not only a kind of physical but it also psychological as well. Due to identity crisis, a person is having an issue of fear, anxiety, and lack of security. Everything has negative and positive aspects. Newspaper columns, magazine articles, films, made-for-TV movies, television special reports, and talk shows are all public arenas where images of political agenda of their own interest are constructed, debated, and reproduced. From these resources, individuals construct their own conceptions of what is normal and acceptable. This bias affects images in the media, and in turn has a negative effect on public development in a society. This paper investigates the relationship between globalization and cultural war, identity crisis and the role of violence. Objectives: - To determine which type of media plays an important role in shaping perceptions and attitudes of public negatively; - To analyze the impact of globalization on identity crisis, violence and global culture (positive and negative).

Keywords: paradigm shift, globalization, identity crisis, cultural war

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390 Effect of Chemical Fertilizer on Plant Growth-Promoting Rhizobacteria in Wheat

Authors: Tessa E. Reid, Vanessa N. Kavamura, Maider Abadie, Adriana Torres-Ballesteros, Mark Pawlett, Ian M. Clark, Jim Harris, Tim Mauchline

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The deleterious effect of chemical fertilizer on rhizobacterial diversity has been well documented using 16S rRNA gene amplicon sequencing and predictive metagenomics. Biofertilization is a cost-effective and sustainable alternative; improving strategies depends on isolating beneficial soil microorganisms. Although culturing is widespread in biofertilization, it is unknown whether the composition of cultured isolates closely mirrors native beneficial rhizobacterial populations. This study aimed to determine the relative abundance of culturable plant growth-promoting rhizobacteria (PGPR) isolates within total soil DNA and how potential PGPR populations respond to chemical fertilization in a commercial wheat variety. It was hypothesized that PGPR will be reduced in fertilized relative to unfertilized wheat. Triticum aestivum cv. Cadenza seeds were sown in a nutrient depleted agricultural soil in pots treated with and without nitrogen-phosphorous-potassium (NPK) fertilizer. Rhizosphere and rhizoplane samples were collected at flowering stage (10 weeks) and analyzed by culture-independent (amplicon sequence variance (ASV) analysis of total rhizobacterial DNA) and -dependent (isolation using growth media) techniques. Rhizosphere- and rhizoplane-derived microbiota culture collections were tested for plant growth-promoting traits using functional bioassays. In general, fertilizer addition decreased the proportion of nutrient-solubilizing bacteria (nitrate, phosphate, potassium, iron and, zinc) isolated from rhizocompartments in wheat, whereas salt tolerant bacteria were not affected. A PGPR database was created from isolate 16S rRNA gene sequences and searched against total soil DNA, revealing that 1.52% of total community ASVs were identified as culturable PGPR isolates. Bioassays identified a higher proportion of PGPR in non-fertilized samples (rhizosphere (49%) and rhizoplane (91%)) compared to fertilized samples (rhizosphere (21%) and rhizoplane (19%)) which constituted approximately 1.95% and 1.25% in non-fertilized and fertilized total community DNA, respectively. The analyses of 16S rRNA genes and deduced functional profiles provide an in-depth understanding of the responses of bacterial communities to fertilizer; this study suggests that rhizobacteria, which potentially benefit plants by mobilizing insoluble nutrients in soil, are reduced by chemical fertilizer addition. This knowledge will benefit the development of more targeted biofertilization strategies.

Keywords: bacteria, fertilizer, microbiome, rhizoplane, rhizosphere

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389 Sexual Consent and Persons with Psychosocial Disabilities: Exploring Sexual Rights under Indian Laws

Authors: Sachin Sharma

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Sexual consent is integral to every sexual relationship. It is a process to facilitate sexual autonomy and bodily integrity. It assures complete sexual personhood and allows an individual to explore her sexual expressions independently. But the said proposition is not true for people with psychosocial disabilities. Generally, they are considered seraphic or mephistophelic and denied access to sexual autonomy. This result in institutionalizing the sexuality of disabled persons, where the eugenics-ableist narrative defines assessment and access to consent. This way, sexuality and disability are distanced apart. It is primarily due to the stigmatized socio-cultural constructs of sexuality that define sex within a “standard” and “charmed” circle. Such stigmatized expression influences the law, as it considers people with psychosocial disabilities incapable of sexual consent. The approach of legal institutions is very narrow towards interpreting their sexual rights. It echoes the modernist-ableism and strangulates the sexual choices. This way, it reflects the repressive model of sex and denies space to people with psychosocial disabilities. Moreover, judicial courts follow old and conservative methods while dealing with sexual issues. For instance, courts still practice the “standardized” norm of intelligence quotient (IQ) for determining the credibility of persons with psychosocial disabilities. Further, there is still doubt about assistive communicative techniques. This paper will try to question the normative structure of sexual consent and related laws while specifically addressing the issues of sex as desire and abuse. Considering the commitment to the United Nations Convention on the Rights of Persons with Disabilities (herein referred to as UNCRPD) and common law experience, the paper will draw a comparative study on the legal position of sexual rights in India. The paper will also analyze the role of UNCRPD in addressing sexual rights. The author will examine the position of sexual rights of people with psychosocial disabilities after the drafting of UNCRPD and specific state laws. The paper primarily follows the doctrinal method.

Keywords: sexual autonomy, institutionalized choices, overregulated laws, violation of individuality

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388 Effects of Gamma-Tocotrienol Supplementation on T-Regulatory Cells in Syngeneic Mouse Model of Breast Cancer

Authors: S. Subramaniam, J. S. A. Rao, P. Ramdas, K. R. Selvaduray, N. M. Han, M. K. Kutty, A. K. Radhakrishnan

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Immune system is a complex system where the immune cells have the capability to respond against a wide range of immune challenges including cancer progression. However, in the event of cancer development, tumour cells trigger immunosuppressive environment via activation of myeloid-derived suppressor cells and T regulatory (Treg) cells. The Treg cells are subset of CD4+ T lymphocytes, known to have crucial roles in regulating immune homeostasis and promoting the establishment and maintenance of peripheral tolerance. Dysregulation of these mechanisms could lead to cancer progression and immune suppression. Recently, there are many studies reporting on the effects of natural bioactive compounds on immune responses against cancer. It was known that tocotrienol-rich-fraction consisting 70% tocotrienols and 30% α-tocopherol is able to exhibit immunomodulatory as well as anti-cancer properties. Hence, this study was designed to evaluate the effects of gamma-tocotrienol (G-T3) supplementation on T-reg cells in a syngeneic mouse model of breast cancer. In this study, female BALB/c mice were divided into two groups and fed with either soy oil (vehicle) or gamma-tocotrienol (G-T3) for two weeks followed by inoculation with tumour cells. All the mice continued to receive the same supplementation until day 49. The results showed a significant reduction in tumour volume and weight in G-T3 fed mice compared to vehicle-fed mice. Lung and liver histology showed reduced evidence of metastasis in tumour-bearing G-T3 fed mice. Besides that, flow cytometry analysis revealed T-helper cell population was increased, and T-regulatory cell population was suppressed following G-T3 supplementation. Moreover, immunohistochemistry analysis showed that there was a marked decrease in the expression of FOXP3 in the G-T3 fed tumour bearing mice. In conclusion, the G-T3 supplementation showed good prognosis towards breast cancer by enhancing the immune response in tumour-bearing mice. Therefore, gamma-T3 can be used as immunotherapy agent for the treatment of breast cancer.

Keywords: breast cancer, gamma tocotrienol, immune suppression, supplement

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387 A Research on a Historical Architectural Heritage of the Village: Zriba El Olia

Authors: Yosra Ben Salah, Wang Li Jun, Salem Bellil

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The village Hammem Zriba is a lost little paradise in the middle of a beautiful landscape that captures the eyes of every visitor. The village alone is a rich expression of different elements such as urban, architecture, technical and vernacular elements, as well as sociological, spiritual and religious behaviors. This heritage is in degrading conditions and is threatened by disappearing soon; thus, actions have to be taken as soon as possible to preserve this heritage, record, analyze and learn from its traditional ways of construction. The strategy of this study is to examine the architecture within the Berber society over a period of time and influenced by a certain location and its relationship to the social and cultural aspects; this research will focus on historical, environmental, social and cultural aspects influencing architecture. The contents of this paper should mainly be constructed by three successive layouts of historical view, a cultural view and an architectural view that will include the urban and domestic scale. This research relies on the integration of both theoretical and empirical investigations. On the theoretical level: A documentary analysis of secondary data is used. Documentary analysis means content analysis of the relevant documents that include books, journals, magazines, archival data, and field survey and observations. On the empirical level: analysis of these traditional ways of planning and house building will be carried out. Through the Analysis, three techniques will be employed to collect primary data. These techniques are; systematic analysis of the architectural drawings, quantitative analysis to the houses statistics, and a direct observation. Through this research, the technical, architectural and urban achievements of the Berber people who represent a part of the general history and architectural history will be emphasized. And on a second point the potential for the sustainability present in this traditional urban planning and housing to be used to formulate guidelines for modern urban and housing development.

Keywords: culture, history, traditional architecture, values

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386 A Systems Approach to Targeting Cyclooxygenase: Genomics, Bioinformatics and Metabolomics Analysis of COX-1 -/- and COX-2-/- Lung Fibroblasts Providing Indication of Sterile Inflammation

Authors: Abul B. M. M. K. Islam, Mandar Dave, Roderick V. Jensen, Ashok R. Amin

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A systems approach was applied to characterize differentially expressed transcripts, bioinformatics pathways, and proteins and prostaglandins (PGs) from lung fibroblasts procured from wild-type (WT), COX-1-/- and COX-2-/- mice to understand system level control mechanism. Bioinformatics analysis of COX-2 and COX-1 ablated cells induced COX-1 and COX-2 specific signature respectively, which significantly overlapped with an 'IL-1β induced inflammatory signature'. This defined novel cross-talk signals that orchestrated coordinated activation of pathways of sterile inflammation sensed by cellular stress. The overlapping signals showed significant over-representation of shared pathways for interferon y and immune responses, T cell functions, NOD, and toll-like receptor signaling. Gene Ontology Biological Process (GOBP) and pathway enrichment analysis specifically showed an increase in mRNA expression associated with: (a) organ development and homeostasis in COX-1-/- cells and (b) oxidative stress and response, spliceosomes and proteasomes activity, mTOR and p53 signaling in COX-2-/- cells. COX-1 and COX-2 showed signs of functional pathways committed to cell cycle and DNA replication at the genomics level. As compared to WT, metabolomics analysis revealed a significant increase in COX-1 mRNA and synthesis of basal levels of eicosanoids (PGE2, PGD2, TXB2, LTB4, PGF1α, and PGF2α) in COX-2 ablated cells and increase in synthesis of PGE2, and PGF1α in COX-1 null cells. There was a compensation of PGE2 and PGF1α in COX-1-/- and COX-2-/- cells. Collectively, these results support a broader, differential and collaborative regulation of both COX-1 and COX-2 pathways at the metabolic, signaling, and genomics levels in cellular homeostasis and sterile inflammation induced by cellular stress.

Keywords: cyclooxygenases, inflammation, lung fibroblasts, systemic

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385 Suicide Attempts and Gender: A Qualitative Analysis in Cuba

Authors: Alejandro Arnaldo Barroso Martinez

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Unlike sex, which is constituted by anatomic-physiological differences, gender is a social construction. Our thoughts and behaviors as females and males are not etched in stone by our biology but rather from how society expects us to think and behave based on our sex assignment in the womb. Social expectations, values, and roles are taken on by individuals and shape the ways considered acceptable and linked to our bodies, feelings, and interpersonal relationships. Furthermore, these evolve into dire consequences for those who do not meet these disciplinary, economic, and cultural standards. Then, the social learning of gender identity implies the individual’s psychological sense of being, and it might be highly linked to a sense of life and suicide attempts. As a result, suicide has been considered a gender issue with differences in the rates and means used by men and women worldwide. Nevertheless, there has been a misunderstanding of the meaning of being male or female in a particular context and how it becomes a risk process for suicide attempts. For this reason, the general objective of the current research is to explain how this process occurs in Cuba. From a Critical Sociology and Social Psychology, a qualitative methodology was developed through six case studies and qualitative in-depth interviews. The analysis is focused on the sequence and interplay between two dimensions of meaning: signifiers and voices. Findings show that the risk process of suicide attempts in Cuba means some patriarchal beliefs and practices as part of informal educational models and some positivist practices in mental health attention. Findings also show that community relations create a sense of belonging, and it is a protection against suicide attempts in Cuba. Those frames of signifiers and voices explain in both males and females but differently when and how they are suffering from isolation, violence, the normalization of emotional awareness, and emotional distress expression. Suicide prevention programs should take gender learning into account as a cultural process.

Keywords: social constructions, gender identity, meanings, suicide attempt

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384 East West Discourse: An Esoteric Comparison of the Western Philosophy and the Eastern Vedanta

Authors: Chandrabati Chakraborty

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The progressive emergence, in the course of the evolution of life, mind and personality, requires us to assume a creative Principle operating timeless Reality in the temporal. The difference between Western philosophy and that of India, concerns the origin and the purpose of the philosophical enquiry. While the former wonders at the external world, the latter is awareness of perennial suffering associated with human existence. The present world suffers from a basic form of rootlessness,reflecting many psychological, philosophical studies. Alienation,a major theme of human condition in the contemporary epoch has emerged as natural consequences of existential predicament. As Edmund Fuller also observes that individuals suffer not only from famine, ruin or even war but also from devastating inner problems, which are that of estrangement, hopelessness and utter despair. This existentialism is thus considered by Jean Wahl as the “Philosophies of existence”.The post world war scenario well analyses the chaos,annihilation,frustration and anguished estrangement. In such conditions when the West cries out , “What is there?I know first of all that I am.But who am I?.....I am sepeated.What I am seperated from I cannot name it. But I am seperated.”(Dostoevsky:The Confession), Vedantic philosophy looks upon the Pilgrim’s Progress of Humanity as being essentially one,operationg squarely within the bounds of reality, reflecting a basic human experience, outbraving indecorous dictims that have failed to give due honour to human beings,echoing for centuries the Sanskrit slokas with ultimate certitude: II Esa Atma samaha plusina samo masakena samo nagena sama ebhis tribhir lokaih.....sama nena sarvena II (The Atman (Divine Soul) is the same in the ant, the same in the gnat, the same in the elephant, the same in these three worlds....the same in the whole Universe). The present paper aims at a comparative study of cultural and philosophical expression taking into view extensive illustrations from Western Philosophers and The Vedantic,Upanishadic lores of Indian philosophy.

Keywords: existentialism, Vedanta, philosophy, absurdism

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383 Relationship of Epidermal Growth Factor Receptor Gene Mutations Andserum Levels of Ligands in Non-Small Cell Lung Carcinoma Patients

Authors: Abdolamir Allameh, Seyyed Mortaza Haghgoo, Adnan Khosravi, Esmaeil Mortaz, Mihan Pourabdollah-Toutkaboni, Sharareh Seifi

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Non-Small Cell Lung Carcinoma (NSCLC) is associated with a number of gene mutations in epidermal growth factor receptor (EGFR). The prognostic significance of mutations in exons 19 and 21, together with serum levels of EGFR, amphiregulin (AR), and Transforming Growth Factor-alpha (TGF-α) are implicated in diagnosis and treatment. The aim of this study was to examine the relationship of EGFR mutations in selected exons with the expression of relevant ligands in sera samples of NSCLC patients. For this, a group of NSCLC patients (n=98) referred to the hospital for lung surgery with a mean age of 59±10.5 were enrolled (M/F: 75/23). Blood specimen was collected from each patient. Besides, formalin fixed paraffin embedded tissues were processed for DNA extraction. Gene mutations in exons 19 and 21 were detected by direct sequencing, following DNA amplification which was done by PCR (Polymerase Chain Reaction). Also, serum levels of EGFR, AR, and TGF-α were measured by ELISA. The results of our study show that EGFR mutations were present in 37% of Iranian NSCLC patients. The most frequently identified mutations were deletions in exon 19 (72.2%) and substitutions in exon 21 (27.8%). The most frequently identified alteration, which is considered as a rare mutation, was the E872K mutation in exon 21, which was found in 90% (9 out of 10) cases. EGFR mutation detected in exon 21 was significantly (P<0.05) correlated with the levels of its ligands, EGFR and TGF-α in serum samples. Furthermore, it was found that increased serum AR (>3pg/ml) and TGF-α (>10.5 pg/ml) were associated with shorter overall survival (P<0.05). The results clearly showed a close relationship between EGFR mutations and serum EGFR and serum TGF-α. Increased serum EGFR was associated with TGF-α and AR and linked to poor prognosis of NSCLC. These findings are implicated in clinical decision-making related to EGFR-Tyrosine kinase inhibitors (TKIs).

Keywords: lung cancer, Iranian patients, epidermal growth factor, mutation, prognosis

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382 Towards an Eastern Philosophy of Religion: on the Contradictory Identity of Philosophy and Religion

Authors: Carlo Cogliati

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The study of the relationship of philosophical reason with the religious domain has been very much a concern for many of the Western philosophical and theological traditions. In this essay, I will suggest a proposal for an Eastern philosophy of religion based on Nishida’s contradictory identity of the two: philosophy soku hi (is, and yes is not) religion. This will pose a challenge to the traditional Western contents and methods of the discipline. This paper aims to serve three purposes. First, I will critically assess Charlesworth’s typology of the relation between philosophy and religion in the West: philosophy as/for/against/about/after religion. I will also engage Harrison’s call for a global philosophy of religion(s) and argue that, although it expands the scope and the range of the questions to address, it is still Western in its method. Second, I will present Nishida’s logic of absolutely contradictory self-identity as the instrument to transcend the dichotomous pair of identity and contradiction: ‘A is A’ and ‘A is not A’. I will then explain how this ‘concrete’ logic of the East, as opposed to the ‘formal’ logic of the West, exhibits at best the bilateral dynamic relation between philosophy and religion. Even as Nishida argues for the non-separability of the two, he is also aware and committed to their mutual non-reducibility. Finally, I will outline the resulting new relation between God and creatures. Nishida in his philosophy soku hi religion replaces the traditional Western dualistic concept of God with the Eastern non-dualistic understanding of God as “neither transcendent nor immanent, and at the same time both transcendent and immanent.” God is therefore a self-identity of contradiction, nowhere and yet everywhere present in the world of creatures. God as absolute being is also absolute nothingness: the world of creatures is the expression of God’s absolute self-negation. The overreaching goal of this essay is to offer an alternative to traditional Western approaches to philosophy of religion based on Nishida’s logic of absolutely contradictory self-identity, as an example of philosophical and religious counter(influence). The resulting relationship between philosophy and religion calls for a revision of traditional concepts and methods. The outcome is not to reformulate the Eastern predilection to not sharply distinguish philosophical thought from religious enlightenment rather to bring together philosophy and religion in the place of identity and difference.

Keywords: basho, Nishida Kitaro, shukyotetsugaku, soku hi, zettai mujunteki jikodoitsu no ronri

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381 Anticancer and Anti-Apoptotic Potential of Tridham and 1,2,3,4,6-Penta-O-Galloyl-β-D-Glucose in MCF-7 Breast Cancer Cell Line

Authors: R. Stalin, D. Karthick, H. Haseena Banu, T. P. Sachidanandam, P. Shanthi

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Background: Breast cancer is emerging as one of the leading cause of cancer related deaths and hence there arises the need to look out for drugs which are more targets specific with minimal side effects. In recent times, there is a shift towards alternative medicine due to low cost and less side effects. Siddha system of medicine is one the oldest system of medicine practiced against various ailments. Tridham (TD) is a herbal formulation prepared in our laboratory consisting of Terminalia chebula, Elaeocarpus ganitrus and Prosopis cineraria in a definite ratio (TD) and its anticancer potential is evaluated in terms of induction of apoptosis. Objective: The present study was designed to investigate the anti proliferative effect of TD and 1,2,3,4,6-penta-O-galloyl-b-D-glucose (PGG), a pure compound isolated from TD on human mammary carcinoma cell line (MCF-7). Materials and Methods: Cell viability was studied using MTT analysis and trypan blue staining. Mitochondrial membrane potential was studied using DAPI staining. The protein and mRNA expressions of pro-apoptotic and anti- apoptotic markers namely Bax, Bad, Bcl-2 and caspases were also assessed by Western Blotting and RT PCR. Results: Viability studies of TD and PGG treated MCF-7 cells showed an inhibition in cell growth in time and dose dependent manner. The alteration in mitochondrial membrane potential was restored through treatment with TD and PGG which was confirmed by DAPI staining. The protein and mRNA expression of pro-apoptotic markers was found to be significantly increased in TD and PGG treated cells with a concomitant decrease in anti-apoptotic markers. Conclusion: The results of the study suggest that TD and PGG exhibit their anticancer effect through its membrane stabilizing property and activation of apoptotic cascade in MCF-7 cells.

Keywords: apoptosis, mammary carcinoma, MCF-7, penta galloyl glucose, Tridham

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380 Neuroprotective Effect of Tangeretin against Potassium Dichromate-Induced Acute Brain Injury via Modulating AKT/Nrf2 Signaling Pathway in Rats

Authors: Ahmed A. Sedik, Doaa Mahmoud Shuaib

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Brain injury is a cause of disability and death worldwide. Potassium dichromate (PD) is an environmental contaminant widely recognized as teratogenic, carcinogenic, and mutagenic towards animals and humans. The aim of the present study was to investigate the possible neuroprotective effects of tangeretin (TNG) on PD-induced brain injury in rats. Forty male adult Wistar rats were randomly and blindly allocated into four groups (8 rats /group). The first group received saline intranasally (i.n.). The second group received a single dose of PD (2 mg/kg, i.n.). The third group received TNG (50 mg/kg; orally) for 14 days, followed by i.n. of PD on the last day of the experiment. Four groups received TNG (100 mg/kg; orally) for 14 days, followed by i.n. of PD on the last day of the experiment. 18- hours after the final treatment, behavioral parameters, neuro-biochemical indices, FTIR analysis, and histopathological studies were evaluated. Results of the present study revealed that rats intoxicated with PD promoted oxidative stress and inflammation via an increase in MDA and a decrease in Nrf2 signaling pathway and GSH levels with an increase in brain contents of TNF-α, IL-10, and NF-kβ and reduced AKT levels in brain homogenates. Treatment with TNG (100 mg/kg; orally) ameliorated behavioral, cholinergic activities and oxidative stress, decreased the elevated levels of pro-inflammatory mediators; TNF-α, IL-10, and NF-κβ elevated AKT pathway with corrected FTIR spectra with a decrease in brain content of chromium residues detected by atomic absorption spectrometry. Also, TNG administration restored the morphological changes as degenerated neurons and necrosis associated with PD intoxication. Additionally, TNG decreased Caspase-3 expression in the brain of PD rats. TNG plays a crucial role in AKT/Nrf2 pathway that is responsible for their antioxidant, anti-inflammatory effects, and apoptotic pathway against PD-induced brain injury in rats.

Keywords: tangeretin, potassium dichromate, brain injury, AKT/Nrf2 signaling pathway, FTIR, atomic absorption spectrometry

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379 Disinformation’s Threats to Democracy in Central Africa: Case Studies from Cameroon and Central African Republic

Authors: Simont Toussi

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Cameroon and the Central African Republic arebound by the provisions of many regional and international charters, which condemn the manipulation of information, obstacles to access reliable information, or the limitation of freedoms of expression and opinion. These two countries also have constitutional guarantees for free speech and access to true and liable information. However, they are yet to define specific policies and regulations for access to information, disinformation, or misinformation. Yet, certain countries’ laws and regulations related to information and communication technologies, to criminal procedures, to terrorism, or intelligence services contain provisions that rather hider human rights by condemning false information. Like many other African countries, Cameroon and the Central African Republic face a profound democratic regression, and governments use multiple methods to stifle online discourse and digital rights. Despite the increased uptake of digital tools for political participation, there is a lack of interactivity and adoption of these tools. This enables a scarcity of information and creates room for the spreading of disinformation in the public space, hamperingdemocracy and the respect for human rights. This research aims to analyse the adequacy of stakeholders’ responses to disinformation in Cameroon and the Central African Republic in periods of political contestation, such as elections and anti-government protests, to highlight the nature, perpetrators, strategies, and channels of disinformation, as well as its effects on democratic actors, including civil society, bloggers, government critics, activists, and other human rights defenders. The study follows a qualitative method with literature review, content analysis, andkey informant’sinterviews with stakeholders’ representatives, emphasized crowdsourcing as a data and information collecting method in the two countries.

Keywords: disinformation, democracy, political manipulation, social media, media, fake news, central Africa, cameroon, misinformation, free speech

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378 Slave Museums and a Site of Democratic Pedagogy: Engagement, Healing and Tolerance

Authors: Elaine Stavro

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In our present world where acts of incivility, intolerance and anger towards minority communities is on the rise, the ways museum practices cultivate ethical generosity is of interest. Democratic theorists differ as to how they believe respect can be generated through active participation. Allowing minority communities a role in determining what artifacts will be displayed and how they will be displayed has been an important step in generating respect. In addition, the rise of indigenous museums, slave museums and curators who represent these communities, contribute to the communication of their history of oppression. These institutional practices have been supplemented by the handling of objects, recognition stories and multisensory exhibitions. Psychoanalysis, object relations theorists believe that the handling of objects: amenable objects and responsive listeners will trigger the expression of anomie, alienation and traumatizing experiences. Not only memorializing but engaging with one’s lose in a very personal way can facilitate the process of mourning. Manchester Museum (UK) gathered together Somalian refugees, who in the process of handling their own objects and those offered at the museum, began to tell their stories. Democratic theorists (especially affect theorists or vital materialists or Actor Network theorists) believe that things can be social actants- material objects have agentic capacities that humans should align with. In doing so, they challenge social constructivism that attributes power to interpreted things, but like them they assume an openness or responsiveness to Otherness can be cultivated. Rich sensory experiences, corporeal engagement (devices that involve bodily movement or objects that involve handling) auditory experiences (songs) all contribute to improve one’s responsiveness and openness to Others. This paper will focus specifically on slave museums/ and exhibits in the U.K, the USA., South Africa to explore and evaluate their democratic strategies in cultivating tolerant practices via the various democratic avenues outlined above.

Keywords: democratic pedagogy, slave exhibitions, affect/emotion, object handling

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377 Online Versus Face-To-Face – How Do Video Consultations Change The Doctor-Patient-Interaction

Authors: Markus Feufel, Friederike Kendel, Caren Hilger, Selamawit Woldai

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Since the corona pandemic, the use of video consultation has increased remarkably. For vulnerable groups such as oncological patients, the advantages seem obvious. But how does video consultation potentially change the doctor-patient relationship compared to face-to-face consultation? Which barriers may hinder the effective use of this consultation format in practice? We are presenting first results from a mixed-methods field study, funded by Federal Ministry of Health, which will provide the basis for a hands-on guide for both physicians and patients on how to improve the quality of video consultations. We use a quasi-experimental design to analyze qualitative and quantitative differences between face-to-face and video consultations based on video recordings of N = 64 actual counseling sessions (n = 32 for each consultation format). Data will be recorded from n = 32 gynecological and n = 32 urological cancer patients at two clinics. After the consultation, all patients will be asked to fill out a questionnaire about their consultation experience. For quantitative analyses, the counseling sessions will be systematically compared in terms of verbal and nonverbal communication patterns. Relative frequencies of eye contact and the information exchanged will be compared using 𝝌2 -tests. The validated questionnaire MAPPIN'Obsdyad will be used to assess the expression of shared decision-making parameters. In addition, semi-structured interviews will be conducted with n = 10 physicians and n = 10 patients experienced with video consultation, for which a qualitative content analysis will be conducted. We will elaborate the comprehensive methodological approach we used to compare video vs. face-to-face consultations and present first evidence on how video consultations change the doctor-patient interaction. We will also outline possible barriers of video consultations and best practices on how they may be overcome. Based on the results, we will present and discuss recommendations outlining best practices for how to prepare and conduct high-quality video consultations from the perspective of both physicians and patients.

Keywords: video consultation, patient-doctor-relationship, digital applications, technical barriers

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376 RAD-Seq Data Reveals Evidence of Local Adaptation between Upstream and Downstream Populations of Australian Glass Shrimp

Authors: Sharmeen Rahman, Daniel Schmidt, Jane Hughes

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Paratya australiensis Kemp (Decapoda: Atyidae) is a widely distributed indigenous freshwater shrimp, highly abundant in eastern Australia. This species has been considered as a model stream organism to study genetics, dispersal, biology, behaviour and evolution in Atyids. Paratya has a filter feeding and scavenging habit which plays a significant role in the formation of lotic community structure. It has been shown to reduce periphyton and sediment from hard substrates of coastal streams and hence acts as a strongly-interacting ecosystem macroconsumer. Besides, Paratya is one of the major food sources for stream dwelling fishes. Paratya australiensis is a cryptic species complex consisting of 9 highly divergent mitochondrial DNA lineages. Among them, one lineage has been observed to favour upstream sites at higher altitudes, with cooler water temperatures. This study aims to identify local adaptation in upstream and downstream populations of this lineage in three streams in the Conondale Range, North-eastern Brisbane, Queensland, Australia. Two populations (up and down stream) from each stream have been chosen to test for local adaptation, and a parallel pattern of adaptation is expected across all streams. Six populations each consisting of 24 individuals were sequenced using the Restriction Site Associated DNA-seq (RAD-seq) technique. Genetic markers (SNPs) were developed using double digest RAD sequencing (ddRAD-seq). These were used for de novo assembly of Paratya genome. De novo assembly was done using the STACKs program and produced 56, 344 loci for 47 individuals from one stream. Among these individuals, 39 individuals shared 5819 loci, and these markers are being used to test for local adaptation using Fst outlier tests (Arlequin) and Bayesian analysis (BayeScan) between up and downstream populations. Fst outlier test detected 27 loci likely to be under selection and the Bayesian analysis also detected 27 loci as under selection. Among these 27 loci, 3 loci showed evidence of selection at a significance level using BayeScan program. On the other hand, up and downstream populations are strongly diverged at neutral loci with a Fst =0.37. Similar analysis will be done with all six populations to determine if there is a parallel pattern of adaptation across all streams. Furthermore, multi-locus among population covariance analysis will be done to identify potential markers under selection as well as to compare single locus versus multi-locus approaches for detecting local adaptation. Adaptive genes identified in this study can be used for future studies to design primers and test for adaptation in related crustacean species.

Keywords: Paratya australiensis, rainforest streams, selection, single nucleotide polymorphism (SNPs)

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375 Pathogenic Escherichia Coli Strains and Their Antibiotic Susceptibility Profiles in Cases of Child Diarrhea at Addis Ababa University, College of Health Sciences, Tikur Anbessa Specialized Hospital, Addis Ababa, Ethiopia

Authors: Benyam Zenebe, Tesfaye Sisay, Gurja Belay, Workabeba Abebe

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Background: The prevalence and antibiogram of pathogenic E. coli strains, which cause diarrhea vary from region to region, and even within countries in the same geographical area. In Ethiopia, diagnostic approaches to E. coli induced diarrhea in children less than five years of age are not standardized. The aim of this study was to determine the involvement of pathogenic E. coli strains in child diarrhea and determine the antibiograms of the isolates in children less than 5 years of age with diarrhea at Addis Ababa University College of Health Sciences TikurAnbessa Specialized Hospital, Addis Ababa, Ethiopia. Methods: A purposive study that included 98 diarrheic children less than five years of age was conducted at Addis Ababa University College of Health Sciences, TikurAnbessa Specialized Hospital, Addis Ababa, Ethiopia to detect pathogenic E. coli biotypes. Stool culture was used to identify presumptive E. coliisolates. Presumptive isolates were confirmed by biochemical tests, and antimicrobial susceptibility tests were performed on confirmed E. coli isolates by the disk diffusion method. DNA was extracted from confirmed isolates by a heating method and subjected to Polymerase Chain Reaction or the presence of virulence genes. Amplified PCR products were analyzed by agarose gel electrophoresis. Data were collected on child demographics and clinical conditions using administered questionnaires. The prevalence of E. coli strains from the total diarrheic children, and the prevalence of pathogenic strains from total E. coli isolates along with their susceptibility profiles; the distribution of pathogenic E.coli biotypes among different age groups and between the sexes were determined by using descriptive statistics. Result: Out of 98 stool specimens collected from diarrheic children less than 5 years of age, 75 presumptive E. coli isolates were identified by culture; further confirmation by biochemical tests showed that only 56 of the isolates were E. coli; 29 of the isolates were found in male children and 27 of them in female children. Out of the 58 isolates of E. coli, 25 pathotypes belonging to different classes of pathogenic strains: STEC, EPEC, EHEC, EAEC were detected by using the PCR technique. Pathogenic E. coli exhibited high rates of antibiotic resistance to many of the antibiotics tested. Moreover, they exhibited multiple drug resistance. Conclusion: This study found that the isolation rate of E. coli and the involvement of antibiotic-resistant pathogenic E. coli in diarrheic children is prominent, and hence focus should be given on the diagnosis and antimicrobial sensitivity testing of pathogenic E. coli at Addis Ababa University College of Health Sciences TikurAnbessa Specialized Hospital. Among antibiotics tested, Cefotitan could be a drug of choice to treat E. coli.

Keywords: antibiotic susceptibility profile, children, diarrhea, E. coli, pathogenic

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374 The Role of Immunologic Diamonds in Dealing with Mycobacterium Tuberculosis; Responses of Immune Cells in Affliction to the Respiratory Tuberculosis

Authors: Seyyed Mohammad Amin Mousavi Sagharchi, Elham Javanroudi

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Introduction: Tuberculosis (TB) is a known disease with hidden features caused by Mycobacterium tuberculosis (MTB). This disease, which is one of the 10 deadliest in the world, has caused millions of deaths in recent decades. Furthermore, TB is responsible for infecting about 30% population of world. Like any infection, TB can activate the immune system by locating and colonization in the human body, especially in the alveoli. TB is granulomatosis, so MTB can absorb the host’s immune cells and other cells to form granuloma. Method: Different databases (e.g., PubMed) were recruited to prepare this paper and fulfill our goals to search and find effective papers and investigations. Results: Immune response to MTB is related to T cell killers and contains CD1, CD4, and CD8 T lymphocytes. CD1 lymphocytes can recognize glycolipids, which highly exist in the Mycobacterial fatty cell wall. CD4 lymphocytes and macrophages form granuloma, and it is the main line of immune response to Mycobacteria. On the other hand, CD8 cells have cytolytic function for directly killing MTB by secretion of granulysin. Other functions and secretion to the deal are interleukin-12 (IL-12) by induction of expression interferon-γ (INF-γ) for macrophages activation and creating a granuloma, and tumor necrosis factor (TNF) by promoting macrophage phagolysosomal fusion. Conclusion: Immune cells in battle with MTB are macrophages, dendritic cells (DCs), neutrophils, and natural killer (NK) cells. These immune cells can recognize the Mycobacterium by various receptors, including Toll-like receptors (TLRs), Nod-like receptors (NLRs), and C-type lectin receptors (CLRs) located in the cell surface. In human alveoli exist about 50 dendritic macrophages, which have close communication with other immune cells in the circulating system and epithelial cells to deal with Mycobacteria. Against immune cells, MTB handles some factors (e.g., cordfactor, O-Ag, lipoarabinomannan, sulfatides, and adenylate cyclase) and practical functions (e.g., inhibition of macrophages).

Keywords: mycobacterium tuberculosis, immune responses, immunological mechanisms, respiratory tuberculosis

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373 Biochemical Characterization and Structure Elucidation of a New Cytochrome P450 Decarboxylase

Authors: Leticia Leandro Rade, Amanda Silva de Sousa, Suman Das, Wesley Generoso, Mayara Chagas Ávila, Plinio Salmazo Vieira, Antonio Bonomi, Gabriela Persinoti, Mario Tyago Murakami, Thomas Michael Makris, Leticia Maria Zanphorlin

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Alkenes have an economic appeal, especially in the biofuels field, since they are precursors for drop-in biofuels production, which have similar chemical and physical properties to the conventional fossil fuels, with no oxygen in their composition. After the discovery of the first P450 CYP152 OleTJE in 2011, reported with its unique property of decarboxylating fatty acids (FA), by using hydrogen peroxide as a cofactor and producing 1-alkenes as the main product, the scientific and technological interest in this family of enzymes vastly increased. In this context, the present work presents a new decarboxylase (OleTRN) with low similarity with OleTJE (32%), its biochemical characterization, and structure elucidation. As main results, OleTRN presented a high yield of expression and purity, optimum reaction conditions at 35 °C and pH from 6.5 to 8.0, and higher specificity for oleic acid. Besides that, structure-guided mutations were performed and according to the functional characterizations, it was observed that some mutations presented different specificity and chemoselectivity by varying the chain-length of FA substrates from 12 to 20 carbons. These results are extremely interesting from a biotechnological perspective as those characteristics could diversify the applications and contribute to designing better cytochrome P450 decarboxylases. Considering that peroxygenases have the potential activity of decarboxylating and hydroxylating fatty acids and that the elucidation of the intriguing mechanistic involved in the decarboxylation preferential from OleTJE is still a challenge, the elucidation of OleTRN structure and the functional characterizations of OleTRN and its mutants contribute to new information about CYP152. Besides that, the work also contributed to the discovery of a new decarboxylase with a different selectivity profile from OleTJE, which allows a wide range of applications.

Keywords: P450, decarboxylases, alkenes, biofuels

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372 Antioxidant Potential of Pomegranate Rind Extract Attenuates Pain, Inflammation and Bone Damage in Experimental Rats

Authors: Ritu Karwasra, Surender Singh

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Inflammation is an important physiological response of the body’s self-defense system that helps in eliminating and protecting organism from harmful stimuli and in tissue repair. It is a highly regulated protective response which helps in eliminating the initial cause of cell injury, and initiates the process of repair. The present study was designed to evaluate the ameliorative effect of pomegranate rind extract on pain and inflammation. Hydroalcoholic standardized rind extract of pomegranate at doses 50, 100 and 200 mg/kg and indomethacin (3 mg/kg) was tested against eddy’s hot plate induced thermal algesia, carrageenan (acute inflammation) and Complete Freund’s Adjuvant (chronic inflammation) induced models in Wistar rats. Parameters analyzed were inhibition of paw edema, measurement of joint diameter, levels of GSH, TBARS, SOD, TNF-α, radiographic imaging, tissue histology and synovial expression of pro-inflammatory cytokine receptor (TNF-R1). Radiological and light microscopical analysis were carried out to find out the bone damage in CFA-induced chronic inflammatory model. Findings of the present study revealed that pomegranate rind extract at a dose of 200 mg/kg caused a significant (p<0.05) reduction in paw swelling in both the inflammatory models. Nociceptive threshold was also significantly (p<0.05) improved. Immunohistochemical analysis of TNF-R1 in CFA-induced group showed elevated level, whereas reduction in level of TNF-R1 was observed in pomegranate (200 mg/kg). Henceforth, we might say that pomegranate produced a dose-dependent reduction in inflammation and pain along with the reduction in levels of oxidative stress markers and tissue histology, and the effect was found to be comparable to that of indomethacin. Thus, it can be concluded that pomegranate is a potential therapeutic target in the pathogenesis of inflammation and pain, and punicalagin is the major constituents found in rind extract might be responsible for the activity.

Keywords: carrageenan, inflammation, nociceptive-threshold, pomegranate, histopathology

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371 2D and 3D Breast Cancer Cells Behave Differently to the Applied Free Palbociclib or the Palbociclib-Loaded Nanoparticles

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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370 Mordechai Vanunu: “The Atomic Spy” as a Nuclear Threat to Discourse in Israeli Society

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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369 Caring and Sustainable Government: An Examination of Political Vision of Jeong Do-Jeon

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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368 Developing Novel Bacterial Primase (DnaG) Inhibitors

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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367 The Administration of Infection Diseases During the Pandemic COVID-19 and the Role of the Differential Diagnosis with Biomarkers VB10

Authors: Sofia Papadimitriou

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INTRODUCTION: The differential diagnosis between acute viral and bacterial infections is an important cost-effectiveness parameter at the stage of the treatment process in order to achieve the maximum benefits in therapeutic intervention by combining the minimum cost to ensure the proper use of antibiotics.The discovery of sensitive and robust molecular diagnostic tests in response to the role of the host in infections has enhanced the accurate diagnosis and differentiation of infections. METHOD: The study used a sample of six independent blood samples (total=756) which are associated with human proteins-proteins, each of which at the transcription stage expresses a different response in the host network between viral and bacterial infections.Τhe individual blood samples are subjected to a sequence of computer filters that identify a gene panel corresponding to an autonomous diagnostic score. The data set and the correspondence of the gene panel to the diagnostic patents a new Bangalore -Viral Bacterial (BL-VB). FINDING: We use a biomarker based on the blood of 10 genes(Panel-VB) that are an important prognostic value for the detection of viruses from bacterial infections with a weighted average AUROC of 0.97(95% CL:0.96-0.99) in eleven independent samples (sets n=898). We discovered a base with a patient score (VB 10 ) according to the table, which is a significant diagnostic value with a weighted average of AUROC 0.94(95% CL: 0.91-0.98) in 2996 patient samples from 56 public sets of data from 19 different countries. We also studied VB 10 in a new cohort of South India (BL-VB,n=56) and found 97% accuracy in confirmed cases of viral and bacterial infections. We found that VB 10 (a)accurately identifies the type of infection even in unspecified cases negative to the culture (b) shows its clinical condition recovery and (c) applies to all age groups, covering a wide range of acute bacterial and viral infectious, including non-specific pathogens. We applied our VB 10 rating to publicly available COVID 19 data and found that our rating diagnosed viral infection in patient samples. RESULTS: Τhe results of the study showed the diagnostic power of the biomarker VB 10 as a diagnostic test for the accurate diagnosis of acute infections in recovery conditions. We look forward to helping you make clinical decisions about prescribing antibiotics and integrating them into your policies management of antibiotic stewardship efforts. CONCLUSIONS: Overall, we are developing a new property of the RNA-based biomarker and a new blood test to differentiate between viral and bacterial infections to assist a physician in designing the optimal treatment regimen to contribute to the proper use of antibiotics and reduce the burden on antimicrobial resistance, AMR.

Keywords: acute infections, antimicrobial resistance, biomarker, blood transcriptome, systems biology, classifier diagnostic score

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366 Unification of Lactic Acid Bacteria and Aloe Vera for Healthy Gut

Authors: Pavitra Sharma, Anuradha Singh, Nupur Mathur

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There exist more than 100 trillion bacteria in the digestive system of human-beings. Such bacteria are referred to as gut microbiota. Gut microbiota comprises around 75% of our immune system. The bacteria that comprise the gut microbiota are unique to every individual and their composition keeps changing with time owing to factors such as the host’s age, diet, genes, environment, and external medication. Of these factors, the variable easiest to control is one’s diet. By modulating one’s diet, one can ensure an optimal composition of the gut microbiota yielding several health benefits. Prebiotics and probiotics are two compounds that have been considered as viable options to modulate the host’s diet. Prebiotics are basically plant products that support the growth of good bacteria in the host’s gut. Examples include garden asparagus, aloe vera etc. Probiotics are living microorganisms that exist in our intestines and play an integral role in promoting digestive health and supporting our immune system in general. Examples include yogurt, kimchi, kombucha etc. In the context of modulating the host’s diet, the key attribute of prebiotics is that they support the growth of probiotics. By developing the right combination of prebiotics and probiotics, food products or supplements can be created to enhance the host’s health. An effective combination of prebiotics and probiotics that yields health benefits to the host is referred to as synbiotics. Synbiotics comprise of an optimal proportion of prebiotics and probiotics, their application benefits the host’s health more than the application of prebiotics and probiotics used in isolation. When applied to food supplements, synbiotics preserve the beneficial probiotic bacteria during storage period and during the bacteria’s passage through the intestinal tract. When applied to the gastrointestinal tract, the composition of the synbiotics assumes paramount importance. Reason being that for synbiotics to be effective in the gastrointestinal tract, the chosen probiotic must be able to survive in the stomach’s acidic environment and manifest tolerance towards bile and pancreatic secretions. Further, not every prebiotic stimulates the growth of a particular probiotic. The prebiotic chosen should be one that not only maintains 2 balance in the host’s digestive system, but also provides the required nutrition to probiotics. Hence in each application of synbiotics, the prebiotic-probiotic combination needs to be carefully selected. Once the combination is finalized, the exact proportion of prebiotics and probiotics to be used needs to be considered. When determining this proportion, only that amount of a prebiotic should be used that activates metabolism of the required number of probiotics. It was observed that while probiotics are active is both the small and large intestine, the effect of prebiotics is observed primarily in the large intestine. Hence in the host’s small intestine, synbiotics are likely to have the maximum efficacy. In small intestine, prebiotics not only assist in the growth of probiotics, but they also enable probiotics to exhibit a higher tolerance to pH levels, oxygenation, and intestinal temperature

Keywords: microbiota, probiotics, prebiotics, synbiotics

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365 Understanding the First Mental Breakdown from the Families’ Perspective Through Metaphors

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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364 Construction of Genetic Recombinant Yeasts with High Environmental Tolerance by Accumulation of Trehalose and Detoxication of Aldehyde

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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