Search results for: Frazer Kirk

Authors: Frazer Kirk, Rohen Skiba, Pankaj Saxena

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The natural history of the QAV is poorly understood due to the exceeding rarity of the condition. Incidence rates vary between 0.00028-1%. Classically patients present with Aortic Regurgitation (AR) between 40-60 years of age experiencing palpitations, chest pain, or heart failure. (1, 2) Echocardiography is the mainstay of diagnosis for this condition; however, given the rarity of this condition, it can easily be overlooked, as demonstrated here. The case report that follows serves as a reminder of the condition to reduce the innate cognitive bias to overlook the diagnosis due to the availability heuristic. Intraoperative photography, echocardiographic and magnetic resonance imaging from this case for reference to demonstrate that while the diagnosis of Aortic regurgitation was recognized early, the valve morphology was underappreciated.

Keywords: quadricuspid aortic valve, cardiac surgery, echocardiography, congenital

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Authors: Frazer Kirk, Matthew Yong, Peter Williams, Andrie Strobel

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Pulmonary valve papillary fibroelastoma is an exceedingly rare pathology. The experience and literature regarding them are largely anecdotal and based on sporadic, single case reports. Throughout their known history, two features remain salient that they are classically asymptomatic and found incidentally. The demographic profile of those affected is unclear, as reports regarding those affected are mixed, and there is no clear gender or age predominance, although there is some suggestion of a predisposition to affect females. Nor has there been a well-structured epidemiological study of the entity. Interestingly they are becoming more common on peri-mortum examination. Here-after we describe our experience with a symptomatic presentation of pulmonary papillary fibroelastoma masquerading as a pulmonary embolism and its subsequent assessment and management, with intraoperative photography and echocardiography for reference.

Keywords: cardiac tumor, pulmonary valve, fibroelastoma, cardiac surgery

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Authors: Hudson Akewe

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This research presents complementary results with wider applications on convergence and rate of convergence of classical fixed point theory in Banach spaces to the world of the theory of fixed points of mappings defined in classes of spaces of measurable functions, known in the literature as modular function spaces. The study gives a comprehensive survey of various iterative fixed point results for the classes of multivalued ρ-contractive-like, ρ-quasi-contractive-like, ρ-quasi-contractive, ρ-Zamfirescu and ρ-contraction mappings in the framework of modular function spaces. An example is presented to demonstrate the applicability of the implicit-type iterative schemes to the system of ordinary differential equations. Furthermore, numerical examples are given to show the rate of convergence of the various explicit Kirk-type and implicit Kirk-type iterative schemes under consideration. Our results complement the results obtained on normed and metric spaces in the literature. Also, our methods of proof serve as a guide to obtain several similar improved results for nonexpansive, pseudo-contractive, and accretive type mappings.

Keywords: implicit Kirk-type iterative schemes, multivalued mappings, convergence results, fixed point

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Authors: Kirk Johnson, Gabriel Rubin

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The January 6th Insurrection on the United States Capitol Building has been one of the most disturbing expressions of violence in American history. While there are numerous reasons that led to such aggression, there are destructive politico-religious ideologies and understandings, known as Christian Nationalism, White Christian Nationalism, and Christian Dominionism that influenced the actions of the January 6th insurrectionists. Such ideas are deeply rooted in the unification of church and state, a theocracy, embedded within the understanding that Biblical scripture is the blueprint on which the functioning of the American government should follow. In addition, the concept of racial hierarchy is embedded in politico-religious principles of White Christian Nationalism. This paper will examine how Christian Dominionism is one factor that influenced the January 6th Insurrection.

Keywords: January 6th insurrection, dominionism, Christian nationalism, polygenism

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Authors: Daniel Benson, Yundan Gong, Hannah Kirk

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Advancement in national language processing techniques has led to increased data processing speeds, and reduced the need for cumbersome, manual data processing that is often required when processing data from multilateral organizations for specific purposes. As such, using named entity recognition (NER) modeling and the Organisation of Economically Developed Countries (OECD) Creditor Reporting System database, we present the first geotagged dataset of OECD donor Official Development Assistance (ODA) projects on a global, subnational basis. Our resulting data contains 52,086 ODA projects geocoded to subnational locations across 115 countries, worth a combined $87.9bn. This represents the first global, OECD donor ODA project database with geocoded projects. We use this new data to revisit old questions of how ‘well’ donors allocate ODA to the developing world. This understanding is imperative for policymakers seeking to improve ODA effectiveness.

Keywords: international aid, geocoding, subnational data, natural language processing, machine learning

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Authors: Andrea J. Kirk-Jenkins

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Women in late adolescence and young adulthood are particularly vulnerable to disordered eating, and prior research indicates that those within the college and sorority communities may be especially susceptible. Research has primarily involved comparing eating disorder symptoms between sorority women and non-sorority members using formal eating disorder assessments. This phenomenological study examined sorority members’ (N = 10) perceptions of and lived experiences with various disordered eating behaviors within the sorority culture. Data from individual interviews and photographs indicated two structural themes and 11 textural themes related to factors associated with disordered eating behaviors. These findings point to the existence of both positive and negative aspects of sorority culture, normalization of disordered eating behaviors, and pressure to attain or maintain an ideal body image. Implications for university stakeholders, including college counselors, health center staff, and extracurricular program leaders, are discussed. Further research on the identified textural themes as well as a longitudinal study exploring how perceptions change from rush to alumnae status is suggested.

Keywords: eating disorders, disorder eating behaviors, sorority women, sorority culture, college women

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Authors: Louisa Browne Kirk

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In recent press interviews to promote Portrait de la jeune fille en feu (2019, translated to Portrait of a Lady on Fire in English), director and screenwriter Céline Sciamma and actors Adèle Haenel and Noémie Merlant repeatedly state that they understand the film as (if not uniquely, then unusually) produced via and supportive of ‘the female gaze’. Such a way of seeing stands in opposition to ‘the male gaze’, first theorised by Laura Mulvey as the way in which the female figure is a bearer, not maker, of meaning, a silent signifier through and against whom the male creator/viewer produces his fantasies and obsessions. What, then, is the female gaze? How does a woman produce meaning in and through film? Portrait de la jeune fille en feu and another very recent film, Little Women (2019, directed by Greta Gerwig), are unlikely companion films that understand the female gaze to be the act of one woman looking at another woman, a looking that is mediated through the production of art. In Sciamma’s film this looking is sexual and mediated through painting and in Gerwig’s film looking is familial and mediated through writing. In the schema of these films, art, love, looking and meaning are produced through collaboration. The painted and the painter, the written and the writer, are no longer rendered as subject and object but as dual creators, both always seeing and seen. The gaze of the cinematic woman, mediated through shared artistic practice, is ‘the desire-that-gives’.

Keywords: female gaze, Gerwig, Sciamma, shared artistic practice

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Authors: Linda Frazer

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A pluralistic methodology focuses on promoting an understanding that an alternative juridical framework for the regulation of transnational business negotiations (TBN) between private business parties is fundamentally required. This paper deals with the evolving assessment of the doctoral research of the author which demonstrated that due to insufficient juridical tools, negotiations are commonly misunderstood within the complexity of pluralistic and conflicting legal regimes. This inadequacy causes uncertainty in the enforcement of legal remedies, leaving business parties surprised. Consequently, parties cannot sufficiently anticipate when and how legal rights and obligations are created, often counting on oral or incomplete agreements which may lead to the misinterpretation of the extent of their legal rights and obligations. This uncertainty causes threats to business parties for fear of creating unintended legal obligations or, conversely, that law will not enforce intended agreements for failure to pass the tests of contractual validity. A need to find a manner to set default standards of communications and standards of conduct to monitor our evolving global trade would aid law to provide the security, predictability and foreseeability during alternative dispute resolution required by TBN parties. The conclusion of this study includes a proposal of new trade mechanisms, termed 'Bills of Negotiations' (BON) to enhance party autonomy and promote the ability for TBN parties to self-regulate within the boundaries of law. BON will be guided by a secure juridical institutionalized setting that caters to guiding communications during TBN and resolving disputes that arise along the negotiation processes on a fast track basis.

Keywords: alternative resolution disputes, ADR, good faith, good faith, juridical security, legal regulation, trade mechanisms, transnational business negotiations

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Authors: Laura Gantley, Vanessa M. Conn, Stuart Webb, Kirsty Kirk, Marta Gabryelska, Duncan Holds, Brett W. Stringer, Simon J. Conn

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Trinucleotide repeat disorders comprise ~20 severe, inherited human neuromuscular and neurodegenerative disorders, which are a result of an abnormal expansion of repetitive sequences in the DNA. The most common of these, Huntington’s disease, results from the expansion of the CAG repeat region in exon 1 of the HTT gene via an unknown mechanism. Non-coding RNAs have been implicated in the initiation and progression of many diseases; thus, we focus on one circular RNA (circRNA) molecule arising from non-canonical splicing (back splicing) of HTT pre-mRNA. This circRNA and its mouse orthologue were transgenically overexpressed in human cells (SHSY-5Y and HEK293T) and mouse cells (Mb1), respectively. High-content imaging and flow cytometry demonstrated the overexpression of this circRNA reduces cell proliferation, reduces nuclear size independent of cellular size, and alters cell cycle progression. Analysis of protein by western blot and immunofluorescence demonstrated no change to HTT protein levels but altered nuclear-cytoplasmic distribution without impacting the expansion of the HTT repeat region. As these phenotypic and genotypic changes are found in Huntington’s disease patients, these results may suggest that this circRNA may play a functional role in the progression of Huntington’s disease.

Keywords: cell biology, circular RNAs, Huntington’s disease, molecular biology, neurodegenerative disorders

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Authors: Luz-Marina Roberts, Ancil Kirk, Aisha Donaldson, Anya Seepaul, Jade Lakhan, Shianna Tikasingh

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Climate change has produced a crisis that particularly threatens small island states in the Caribbean. Developers and climate enthusiasts alike are now forced to find new and sustainable ways of building. Focus on existing buildings is particularly needed in Trinidad and Tobago, like other islands, especially as these countries are vulnerable to climate threats and geographic locations with close proximity to a hurricane. Additionally, since many colonial-era style buildings still exist, the idea that they are energy inefficient is at the forefront of the work of policy-makers. The question that remains is can these buildings be retrofitted to reflect the modern era while considering climate resilience. This paper aims to investigate the energy efficiency of colonial-era buildings in Port of Spain and whether these buildings in Trinidad and Tobago, if found to be energy inefficient, can be more energy efficient and sustainable. This involves collecting surveys from building management in colonial-era buildings and researching literature on colonial architecture in the Caribbean and modern innovations in green building designs. Additionally, the data and experiences from the Town and Country Planning Division in the Ministry of Planning and Development of Trinidad and Tobago will inform the paper. This research will aid in re-envisioning how green infrastructure can be applied to urban environments with older buildings and help inform planning policy as it relates to sustainability and energy efficiency.

Keywords: spatial planning, climate resilience, energy efficiency, sustainable development

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Authors: Rochelle Alviz, Kirk Johnson

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This study explores the changing nature of gender roles in two traditional island societies. The research focuses particularly on the settler populations within each community (the Filipinos in Guam and the Javanese in Bali). The research seeks to understand the influence of both the forces of globalization and the dynamics of competing cultures on gender roles. To achieve this, a qualitative research design is used, employing in-depth interviews with individuals from both communities and field notes from participant observation. The study finds that globalization and competing cultural norms have influenced traditional gender roles and expectations in two primary areas of social life: the family and the economy. The importance of these two areas of social life to both communities has led to changes and adaptations in gender roles. In the family context, individuals reconcile their traditional gender roles from their country of origin with the dominant or indigenous gender roles of their new place of residence. In the economic context, the study finds that gender roles influence economic participation, including the types of jobs individuals pursue based on their gender roles. The results of the study provide valuable insights into the complexities and nuances of gender roles and the different factors that influence their evolution and adaptation over time. The research also highlights the influence of globalization on traditional societies and settler populations and the ways in which individuals navigate the competing cultural norms and expectations surrounding gender roles. The research contributes to a better understanding of the interplay between culture, globalization, and gender roles and the implications of these changes for individuals and communities.

Keywords: gender roles, culture, settler communities, family, economy, Guam, Bali, globalization

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Authors: Frazer G. Thompson

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The 1990s in America ended with a mass-school-shooting (at least four killed by gunfire excluding the perpetrator(s)) at Columbine High School in Littleton, Colorado. Post-event, many demanded that government and civilian experts develop a ‘profile’ of the potential school shooter in order to identify and preempt likely future acts of violence. This grounded theory research study seeks to explore the validity of the original hypotheses proposed by the Federal Bureau of Investigation (FBI) in 2000, as it relates to the commonality of disclosure by perpetrators of mass-school-shootings, by evaluating fourteen mass-school-shooting events between 2000 and 2019 at locations around the United States. Methods: The strategy of inquiry seeks to investigate case files, public records, witness accounts, and available psychological profiles of the shooter. The research methodology is inclusive of one-on-one interviews with members of the FBI’s Critical Incident Response Group seeking perspective on commonalities between individuals; specifically, disclosure of intent pre-event. Results: The research determined that school shooters do not ‘unfailingly’ notify others of their plans. However, in nine of the fourteen mass-school-shooting events analyzed, the perpetrator did inform the third party of their intent pre-event in some form of written, oral, or electronic communication. In the remaining five instances, the so-called ‘red-flag’ indicators of the potential for an event to occur were profound, and unto themselves, might be interpreted as notification to others of an imminent deadly threat. Conclusion: Data indicates that conclusions drawn in the FBI’s threat assessment perspective published in 2000 are relevant and current. There is evidence that despite potential ‘red-flag’ indicators which may or may not include a variety of other characteristics, perpetrators of mass-school-shooting events are likely to share their intentions with others through some form of direct or indirect communication. More significantly, implications of this research might suggest that society is often informed of potential danger pre-event but lacks any equitable means by which to disseminate, prevent, intervene, or otherwise act in a meaningful way considering said revelation.

Keywords: columbine, FBI profiling, guns, mass shooting, mental health, school violence

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Authors: Mads Kirk Larsen

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Strict air pollution control is today high on the agenda world-wide. By reducing the particular emission, not only the mg/Nm3 will be reduced – also parts of mercury and other hazardous matters attached to the particles will be reduced. Furthermore, it is possible to catch the fine particles (PM2.5). For particulate control, the precipitators are still the preferred choice and much efforts have been done to improve the efficiencies. Many ESP’s have seen electrical upgrading by changing the traditional 1 phase power system into either 3 phase or SMPS (High Frequency) units. However, there exist a 4th type of power supply – the pulse type. This is unfortunately widely unknown, but may be of great benefit to power plants. The FLSmidth type is called COROMAX® and it is a high voltage pulse generator for precipitators using a semiconductor switch operating at medium potential. The generated high voltage pulses have rated amplitude of 80 kV and duration of 75 μs and are superimposed on a variable base voltage of 60 kV rated voltage. Hereby, achieving a peak voltage of 140 kV. COROMAX® has the ability to increase the voltage beyond the natural spark limit inside the precipitator. Voltage levels may often be twice as high after installation of COROMAX®. Hereby also the migration velocity increases and thereby the efficiency. As the collection efficiency is proportional to the voltage peak and mean values, this also increases the collection efficiency of the fine particles where test has shown 80% removal of particles less than 0.07 micron. Another great advantage is the indifference to back-corona. Simultaneously with emission reduction, the power consumption will also be reduced. Another great advantage of the COROMAX® system is that the emission can be improved without the need to change the internal parts or enlarge the ESP. Recently, more than 150 units have been installed in China, where emissions have been reduced to ultra-low levels.

Keywords: eleectrostatic precipitator, high resistivity dust, micropulse energization, particulate removal

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Authors: Christopher B. Obida, George A. Blackburn, James D. Whyatt, Kirk T. Semple

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In many regions of the world, especially in developing countries, river network data are outdated or completely absent, yet such information is critical for supporting important functions such as flood mitigation efforts, land use and transportation planning, and the management of water resources. In this study, a method was developed for delineating river networks using Sentinel 1 imagery. Unsupervised classification was applied to multi-temporal Sentinel 1 data to discriminate water bodies from other land covers then the outputs were combined to generate a single persistent water bodies product. A thinning algorithm was then used to delineate river centre lines, which were converted into vector features and built into a topologically structured geometric network. The complex river system of the Niger Delta was used to compare the performance of the Sentinel-based method against alternative freely available water body products from United States Geological Survey, European Space Agency and OpenStreetMap and a river network derived from a Shuttle Rader Topography Mission Digital Elevation Model. From both raster-based and vector-based accuracy assessments, it was found that the Sentinel-based river network products were superior to the comparator data sets by a substantial margin. The geometric river network that was constructed permitted a flow routing analysis which is important for a variety of environmental management and planning applications. The extracted network will potentially be applied for modelling dispersion of hydrocarbon pollutants in Ogoniland, a part of the Niger Delta. The approach developed in this study holds considerable potential for generating up to date, detailed river network data for the many countries where such data are deficient.

Keywords: Sentinel 1, image processing, river delineation, large scale mapping, data comparison, geometric network

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Authors: Shah Awan, Fahad Sultan, Shahid Jan Kakakhel

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Higher education in the 21st century has been faced with game-changing developments impacting teaching and learning and also strengthening the academia and industry relationship. The academia and industry relationship plays a key role in economic development in developed, developing and emerging economies. The partnership not only explores innovation but also provide a real time experience of the theoretical knowledge. For this purpose, the paper assessing the role of HEC in the Pakistan and discusses the way in academia and industry contribute their role in improving Pakistani economy. Successive studies have reported the importance of innovation and technology , research development initiatives in public sector universities, and the significance of role of higher education commission in strengthening the academia and industrial relationship to improve performance and minimize failure. The paper presents the results of interviews conducted, using semi-structured interviews amongst 26 staff members of two public sector universities, higher education commission and managers from corporate sector.The study shows public sector universities face the several barriers in developing economy like Pakistan, to establish the successful collaboration between universities and industry. Of the participants interviewed, HEC provides an insufficient road map to improve organisational capabilities in facilitating and enhance the performance. The results of this study have demonstrated that HEC has to embrace and internalize support to industry and public sector universities to compete in the era of globalization. Publication of this research paper will help higher education sector to further strengthen research sector through industry and university collaboration. The research findings corroborate the findings of Dooley and Kirk who highlights the features of university-industry collaboration. Enhanced communication has implications for the quality of the product and human resource. Crucial for developing economies, feasible organisational design and framework is essential for the university-industry relationship.

Keywords: higher education commission, role, academia and industry relationship, Pakistan

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Authors: Andrea Parisi, Samantha Vilkins, Luis Furuya-Kanamori, John A. Crump, Benjamin P. Howden, Darren Gray, Kathryn Glass, Martyn Kirk

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Objectives: Salmonella is a leading cause of foodborne enterocolitis worldwide. Nontyphoidal Salmonella (NTS) infections that are Multi-Drug Resistant (MDR) (non-susceptible to ≥1 agent in ≥3 antimicrobial categories) may result in more severe outcomes, although these effects have not been systematically examined. We conducted a systematic review and meta-analysis to examine impacts of MDR NTS on health in high-income settings. Methods: We systematically reviewed the literature from scientific databases, including PubMed, Scopus and grey literature sources, using PRISMA guidelines. We searched for data from case-control studies, cohorts, outbreaks, reports and theses, imposing no language restriction. We included only publications from January 1990 to September 2016 from high income countries as classified by World Bank. We extracted data from papers on duration of illness, hospitalisation rates, morbidity and mortality for MDR and non-MDR NTS strains. Results: After removing duplicates, the initial search revealed 4258 articles. After further screening, we identified 16 eligible studies for the systematic review, and 9 of these were included in meta-analysis. NTS serotypes differed among the reported studies but serotype Typhimurium, Enteritidis, Newport and Heidelberg were among the most often reported as MDR pathogens. Salmonella infections that were MDR were associated with excess bloodstream infections (OR 1.63; 95%CI 1.18-2.26), excess hospitalisations (OR 2.77; 95%CI 1.47-5.21) and higher mortality (OR 3.54; 95%CI 1.10-11.40). Conclusions: MDR NTS infections are a serious public health concern. With the emergence of MDR Salmonella strains in the high-income countries, it is crucial to restrict the use of antimicrobials both in animals and humans, and intervene to prevent foodborne infections.

Keywords: Antimicrobial Resistance, Bloodstream Infection, Health Outcomes, Hospitalisation, Invasive Disease, Multi-Drug Resistance (MDR), Mortality, Nontyphoidal Salmonella

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Authors: Frazer McDonald Ng'oma

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Online Learning in Malawi has only immersed in recent years due to the need to increase access to higher education, the need to accommodate upgrading students who wish to study on a part time basis while still continuing their work, and the COVID-19 pandemic, which forced the closure of schools resulting in academic institutions seeking alternative modes of teaching and Learning to ensure continued teaching and Learning. Realizing that this mode of Learning is becoming a norm, institutions of higher Learning have started pioneering online post-graduate programs from which they can draw lessons before fully implementing it in undergraduate programs. Online learning pedagogy has not been fully grasped and institutions are still experimenting with this mode of Learning until online Learning guiding policies are created and its standards improved. This single case descriptive qualitative research study sought to investigate online postgraduate students’ perceptions and experiences with Student to student interactive pedagogy in their programs. The results of the study are to inform institutions and educators how to structure their programs to ensure that their students get the full satisfaction. 25 Masters students in 3 recently introduced online programs at Kamuzu University of Health Sciences (KUHES), were engaged; 19 were interviewed and 6 responded to questionnaires. The findings from the students were presented and categorized in themes and subthemes that emerged from the qualitative data that was collected and analysed following Colaizzi’s framework for data analysis that resulted in themes formulation. Findings revealed that Student to student interactions occurred in the online programme during live sessions, on class Whatsapp group, in discussion boards as well as on emails. Majority of the students (n=18) felt the level of students’ interaction initiated by the institution was too much, referring to mandatory interactions activities like commenting in discussion boards and attending to live sessons. Some participants (n=7) were satisfied with the level of interaction and also pointed out that they would be fine with more program-initiated student–to–student interactions. These participants attributed having been out of school for some time as a reason for needing peer interactions citing that it is already difficult to get back to a traditional on-campus school after some time, let alone an online class where there is no physical interaction with other students. In general, majority of the participants (n=18) did not value Student to student interaction in online Learning. The students suggested that having intensive student-to-student interaction in postgraduate online studies does not need to be a high priority for the institution and they further recommended that if a lecturer decides to incorporate student-to-student activities into a class, they should be optional.

Keywords: online learning, interactions, student interactions, post graduate students

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Authors: Frazer G. Thompson

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Israel is a nation of cultural and historical diversity, yet the success factors for a modern Bedouin-Arab high-tech entrepreneur seem to be different from those of other Jewish-Israeli citizens. The purpose of this descriptive narrative case study is to explore how an Arab-Israeli all Negev-Bedouin technology company has succeeded in the Israeli high-tech industry by utilizing technology and engineering career opportunities available to Bedouin youth for ‘Sadel Tech,’ at Be’er-Sheva, the Negev, Israel. Methods: The strategy of inquiry seeks to explore real-life contextual understandings, multi-level perspectives, and the cultural influences of personal, community, educational, and entrepreneurial factors. The research methodology includes in-depth one-on-one interviews, focus group sessions, and overt observation to explore the meaning and understanding of the constructs toward determining the effect all or a few of the elements may have on the overall success factors of the company. Results: Study results indicate that the state-run educational system in Israel fails to adequately integrate important aspects of Bedouin culture into the learning environment. However, Bedouin entrepreneurs are finding ways to compensate for these inadequacies by utilizing non-traditional methods of teaching, learning, and doing business. Government incentives for Bedouin start-ups are also recognized as contributors. Employees of Sadel live and work in the Negev, the Gaza Strip, and the West Bank, further informing the study that the traditions of tribal etiquette continue to contribute to modern Bedouin-Arab business culture. Conclusion: Bedouin's business success in Israel is a multi-dimensional concept. While cultural acumen plays a prominent and unique role for both Arab-Israelis and Jewish-Israelis in economic and entrepreneurial pursuits, the marginalization of the Bedouin continues to contribute to the lack of educational and professional opportunities for Bedouin in Israel. Although recognized as important at the government level, programs necessary to implement the infrastructure required to support Bedouin entrepreneurship in Israel remain infantile. The Israeli Government is providing opportunities through grants and other incentives for Bedouin entrepreneurial start-ups, indicating that Israel has recognized the impact of this growing demographic. However, although many Bedouin graduates from University each year with advanced degrees, opportunities for Bedouin within the Israeli high-tech sector remain scarce.

Keywords: Bedouin education, Bedouin entrepreneur, economic anthropology, ethnic business opportunities, Israeli tech, Silicon Wadi

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Authors: Leander Van Neste, Kirk Wojno

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The innate immune system reacts to foreign insult in several unique ways, one of which is phagocytosis of perceived threats such as cancer, bacteria, and viruses. The goal of this study was to look for evidence of phagocytosed RNA from tumor cells in circulating monocytes. While all monocytes possess phagocytic capabilities, the non-classical CD14+/FCGR3A+ monocytes and the intermediate CD14++/FCGR3A+ monocytes most actively remove threatening ‘external’ cellular materials. Purified CD14-positive monocyte samples from fourteen patients recently diagnosed with clinically localized prostate cancer (PCa) were investigated by single-cell RNA sequencing using the 10X Genomics protocol followed by paired-end sequencing on Illumina’s NovaSeq. Similarly, samples were processed and used as controls, i.e., one patient underwent biopsy but was found not to harbor prostate cancer (benign), three young, healthy men, and three men previously diagnosed with prostate cancer that recently underwent (curative) radical prostatectomy (post-RP). Sequencing data were mapped using 10X Genomics’ CellRanger software and viable cells were subsequently identified using CellBender, removing technical artifacts such as doublets and non-cellular RNA. Next, data analysis was performed in R, using the Seurat package. Because the main goal was to identify differences between PCa patients and ‘control’ patients, rather than exploring differences between individual subjects, the individual Seurat objects of all 21 patients were merged into one Seurat object per Seurat’s recommendation. Finally, the single-cell dataset was normalized as a whole prior to further analysis. Cell identity was assessed using the SingleR and cell dex packages. The Monaco Immune Data was selected as the reference dataset, consisting of bulk RNA-seq data of sorted human immune cells. The Monaco classification was supplemented with normalized PCa data obtained from The Cancer Genome Atlas (TCGA), which consists of bulk RNA sequencing data from 499 prostate tumor tissues (including 1 metastatic) and 52 (adjacent) normal prostate tissues. SingleR was subsequently run on the combined immune cell and PCa datasets. As expected, the vast majority of cells were labeled as having a monocytic origin (~90%), with the most noticeable difference being the larger number of intermediate monocytes in the PCa patients (13.6% versus 7.1%; p<.001). In men harboring PCa, 0.60% of all purified monocytes were classified as harboring PCa signals when the TCGA data were included. This was 3-fold, 7.5-fold, and 4-fold higher compared to post-RP, benign, and young men, respectively (all p<.001). In addition, with 7.91%, the number of unclassified cells, i.e., cells with pruned labels due to high uncertainty of the assigned label, was also highest in men with PCa, compared to 3.51%, 2.67%, and 5.51% of cells in post-RP, benign, and young men, respectively (all p<.001). It can be postulated that actively phagocytosing cells are hardest to classify due to their dual immune cell and foreign cell nature. Hence, the higher number of unclassified cells and intermediate monocytes in PCa patients might reflect higher phagocytic activity due to tumor burden. This also illustrates that small numbers (~1%) of circulating peripheral blood monocytes that have interacted with tumor cells might still possess detectable phagocytosed tumor RNA.

Keywords: circulating monocytes, phagocytic cells, prostate cancer, tumor immune response

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