Search results for: Amelia Clarke

Authors: Whitehead-Clarke T., Beynon V., Banks J., Karanjia R., Mudera V., Windsor A., Kureshi A.

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Introduction: Mesh implants are regularly used to help repair both hiatus hernias (HH) and diaphragmatic hernias (DH). In vivo studies are used to test not only mesh safety but increasingly comparative efficacy. Our work examines the field of in vivo mesh testing for HH and DH models to establish current practices and standards. Method: This systematic review was registered with PROSPERO. Medline and Embase databases were searched for relevant in vivo studies. 44 articles were identified and underwent abstract review, where 22 were excluded. 4 further studies were excluded after full text review – leaving 18 to undergo data extraction. Results: Of 18 studies identified, 9 used an in vivo HH model and 9 a DH model. 5 studies undertook mechanical testing on tissue samples – all uniaxial in nature. Testing strip widths ranged from 1-20mm (median 3mm). Testing speeds varied from 1.5-60mm/minute. Upon histology, the most commonly assessed structural and cellular factors were neovascularization and macrophages, respectively (n=9 each). Structural analysis was mostly qualitative, where cellular analysis was equally likely to be quantitative. 11 studies assessed adhesion formation, of which 8 used one of four scoring systems. 8 studies measured mesh shrinkage. Discussion: In vivo studies assessing mesh for HH and DH repair are uncommon. Within this relatively young field, we encourage surgical and materials testing institutions to discuss its standardisation.

Keywords: hiatus, diaphragmatic, hernia, mesh, materials testing, in vivo

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Authors: Mutshinye Manguvhewa, Maria Florence, Mansoo Yu, Elize Koch, Kamal Kamaloodien

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Substance use among pregnant women is a perennial problem in the Western Cape Province of South Africa. There are many influential factors are associated with substance use among women of childbearing age. The study explored factors associated with substance use among pregnant women using a qualitative research design and the bio-ecological theoretical framework to explore and guide the researcher throughout the study. Participants were selected using purposive sampling. Only participants accessed from the Department of Social Development meeting the inclusion criteria of the study were interviewed using semi structured interviews. Immediate referral for psychological intervention during the interview was available for participants who needed it. Braun and Clarke's six phases of thematic analysis were utilised to analyse the data. The study adheres to ethical guidelines for the participants' protection. Participants were informed about the study before the initiation of the interviews and the details of their voluntary participation were explained. The key findings from this study illustrate that socio-cultural factors, personal factors, emotional response and intimate relationships are the major contributing factors to substance use among pregnant women in this sample. The results outline the preventative measures that pregnant women implement. Lastly, the study reveals the positive and negative perceptions of substance use programmes that participants share. Some of the study findings are similar to the existing literature and some of the findings differed. Recommendations emanating from the study include that the stakeholders, rehabilitation centres, Department of Health and future researchers should act proactively against substance use during pregnancy.

Keywords: substance addiction, antenatal care, pregnancy, substance use

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Authors: Marwa Abdussalam

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Breastfeeding is a unique emotional bond between a mother and their offspring. Though breastfeeding may be natural, it is not something mothers are born with; some still struggle to breastfeed their babies. Various factors can influence the breastfeeding experience, such as the mode of delivery, the mother’s health condition, proper latching, etc. In addition, psychological factors have been known to influence breastfeeding ability, duration, and milk supply. Some mothers struggle to breastfeed their babies because they perceive they have a low milk supply and or don’t have the ability to breastfeed their babies. Most of these perceptions result either from their own past experience or from the ‘comments’ of their caregivers. So, it is of utmost essential to understand such psychological factors affecting breastfeeding so that necessary steps can be taken to educate breastfeeding mothers. The study explored the role of psychological factors that affect breastfeeding. Data were collected from fifteen breastfeeding mothers using a semi-structured interview schedule. A total of 10 questions were included in the interview schedule. Questions were sequenced in a funnel pattern, beginning with open-ended questions and then moving on to close-ended questions. Data were analyzed using Braun and Clarke’s Thematic Analysis technique. This technique involves identifying the codes, generating themes, naming them, and finally reviewing them. Results indicated that breastfeeding self-efficacy perceived insufficient milk supply, and lack of knowledge were the psychological factors affecting breastfeeding. The results of this study can be used to help mothers who are struggling with breastfeeding by developing interventions aimed at improving breastfeeding self-efficacy.

Keywords: breastfeeding, breastfeeding self-efficacy, perceived insufficient milk supply, Thematic Analysis

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Authors: Klara Królewiak-Detsi, Anna Orylska, Anna Gorgolewska, Marta Boczkowska, Agata Graczykowska

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This study investigates the communication between students and teachers in inclusive education in Poland. Specifically, we examine the communication and interaction of students with special educational needs during online learning compared to traditional face-to-face instruction. Our research questions are (1) how children with special educational needs communicate with their teachers and peers during online learning, and (2) what strategies can improve their communication skills. We conducted five focus groups with: (1) 55 children with special educational needs, (2) 65 typically developing pupils, (3) 28 professionals (psychologists and special education therapists), (4) 16 teachers, and (5) 16 parents of children with special educational needs. Our analysis focused on primary schools and used thematic analysis according to the 6-step procedure of Braun and Clarke. Our findings reveal that children with disabilities faced more difficulties communicating and interacting with others online than in face-to-face lessons. The online tools used for education were not adapted to the needs of children with disabilities, and schools lacked clear guidelines on how to pursue inclusive education online. Based on the results, we offer recommendations for online communication training and tools that are dedicated to children with special educational needs. Additionally, our results demonstrate that typically developing pupils are better in interpersonal relations and more often and effectively use social support. Children with special educational needs had similar emotional and communication challenges compared to their typically developing peers. In conclusion, our study highlights the importance of providing adequate support for the online education of children with special educational needs in inclusive classrooms.

Keywords: Inclusive education, Special educational needs, Social skills development, Online communication

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Authors: Jayant Balaji Athavale, Sean Clarke

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Background: According to the WHO, every year, more than 700,000 people die by suicide, which is one person around every 45 seconds. Suicide is the fourth leading cause of death among 15 to 29-year-olds globally. The most common situations or life events that might cause suicidal thoughts are financial problems/unemployment, rejections, relationship breakups, sexual/substance abuse and mental illnesses. Mental/psychological weakness caused due to defects in one’s personality is one of the main reasons why people feel they cannot cope in such situations and contemplate suicide. A WHO Mental Health Action Plan 2013–2020 lists a 4-point strategy to enhance mental health by ‘implementing strategies for promotion and prevention in mental health.’ Methodology: With 40 years of spiritual research background, the team at the Maharshi University of Spirituality has studied the spiritual root causes that can significantly affect one’s mental health and the solutions to improve it. Results/Findings: According to spiritual science, the time and nature of death are mostly due to spiritual reasons. A person would mostly contemplate and attempt suicide when he is spiritually most vulnerable. Spiritual practice, as per universal principles, helps in protecting a person spiritually and prevents him from getting such thoughts of self-harm or acting upon them by controlling such impulses. The University has had much success in helping people to overcome the defects in their personalities, including those with suicidal thoughts, through spiritual practices such as chanting the Name of God and the Personality Defect Removal (PDR) process developed by the Author. Conclusion: If such techniques were taught in educational institutions, they could be simple yet effective self-help tools to prevent thoughts of suicide and enhance mental health and well-being.

Keywords: suicide, mental health, abuse, suicide prevention, personality defect removal

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Authors: Fernando Valerio-Holguin

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Over the recent three decades, the tourism industry in the Dominican Republic has had an enormous impact on the country’s culture. The arrival of tourists from Germany, France, Italy, Russia and the United States has rewritten Dominican cultural identity and created a cultural palimpsest in the areas of language, gastronomy, habits, fashion, values, and gender relations. As a consequence of tourism, a prostitution network has flourished across the country. In the film Sand Dollars (2015) directed by Laura Amelia Guzmán and Israel Cárdenas, Noelí (Janet Mojica), a young mulatto woman, altogether with her boyfriend (Ricardo Ariel Toribio), strips tourists of dollars and euro through prostitution. One of her frequent clients is Anne, a mature French woman (Geraldine Chaplin). While Noeli’s goal is to get all the euros she can, Anne falls in love with her and tries to bring her to France. Both the content of the film and its cinematographic languages are analyzed in light of theory of coloniality. This concept shows how European and American tourism, through the power of money, perpetuates colonial discourse, i. e., how race and ethnocentrism permeate cultural activities in their former colonies. Moreover, in the content analysis of the film the concepts of exchange value and fetishism are crucial to understanding how the colonial body becomes sexual commodity. They facilitate grasping the film’s inequity in terms of power in the relationship between the two women: the white old European woman and the young, poor, third-world mulatta. Even though the film attempts to break away from compulsory heterosexuality, the power relation between the two women persists due to the presence of the axis of race, ethnicity, age and gender. Both the novel Les dollars des sables written by Jean-Noel Pancrazi, and the film Sand Dollars offer an interesting insight into sex tourism and coloniality and shed additional light on the power relations between the former colonizers and its colonies.

Keywords: coloniality, ethnocentrism, exchange value, Europe, fetishism, money, power, prostitution, sex tourism, United States of America

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Authors: Samantha Clarke

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The term social justice is central to social work; however, the meaning behind this term is not as simple as defining the term itself. This is because the meaning of social justice is relative since its origin and development is based on evolving political perspectives. Political perspectives provide numerous lenses to view social justice in social work; however, the realities of changing society have meant that social justice has assumed different values, definitions, and understandings over time and in different geopolitical and cultural contexts. There are many competing and convincing theories of social justice that are relevant to social work practice. Exploring the term is not an idle preoccupation because the meaning of the term is not as crucial as the meaning of the worldview, as it is the worldview that positions social justice as crucial in the emancipation of people marginalized from oppression. The many political assumptions that underlie the term social justice are explored and connected to the contemporary discussions about social justice in social work. These connections are then interrogated in the Canadian Social Works Code of Ethics, and in micro, mezzo, and macro approaches. To be remiss in interrogating the underlying political assumptions of the worldview of social justice is to entrench oppression and to preserve oppressive structures in contemporary Canadian social work. The concept of social justice is unable to withstand closer scrutiny about its emancipatory qualities in Canadian social work when we interrogate the many political assumptions that frame its understanding. In order to authenticate social justice as an emancipatory central organizing principle, Canadian social workers must engage in deeper discussions about the political implications of social justice in their everyday practices based on diverse worldviews and geopolitical contexts. Social workers are well positioned to develop an understanding of social justice that is emancipatory based on their everyday practices because as social and political actors they are positioned to work for and with individuals and toward the greater good of those who are marginalized from oppression.

Keywords: Canadian social work, political analysis, social justice, social work practice

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Authors: Myriam Rousseau, Marie-Hélène Poulin, Suzie McKinnon, Jacinthe Bourassa

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Context: There is a growing demand for effective training programs for parents of children with autism spectrum disorders. While traditional in-person training is effective, it can be difficult for some parents to participate due to distance, time, and cost. Telepractice, a form of distance education, could be a viable alternative to address these challenges. Research objective: The objective of this study is to explore the experiences of parents of children with autism who participated in a training program offered by telepractice in order to document: 1) the experience of parents who participated in a program telepractice training program for autistic children, 2) parental satisfaction with the telepractice modality, and 3) potential benefits of using telepractice to deliver training programs to parents of autistic children. Method: This study followed a qualitative research design, and Braun and Clarke's six-step procedure was used for the thematic analysis of the comments provided by parents. Data were collected through individual interviews with parents who participated in the project. The analysis focused on identifying patterns and themes in the comments in order to better understand parents' experiences with the telepractice modality. Results: The study revealed that parents were generally satisfied with the telepractice modality, as it was easy to use and enabled a better balance between work and family. This modality also enabled parents to share and receive mutual support. Despite the positive results, it is still relevant to offer training in different modalities to meet the different needs of parents. Conclusion: The study shows that parents of children with autism are generally satisfied with telepractice as a training modality. The results suggest that telepractice can be an effective alternative to traditional face-to-face training. The study highlights the importance of taking parents' needs and preferences into account when designing and implementing training programs.

Keywords: parents, children, training, telepractice

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Authors: Karish Thavabalan, Alistair Ovenell, Aman Sutaria, Annabelle Parkhouse, Numan Baydemir, Theodore Lally

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Background: Open discussions surrounding menopause are often associated with stigma, with many women feeling uncomfortable to engage in them with friends, colleagues, and healthcare professionals. Though the barriers regarding safe discussions of symptom management experienced by menopausal women are well documented, existing research offers little insight into whether these were shared by pre-menopausal women. This study aimed to explore the barriers regarding safe discussions about menopausal symptom management as perceived or experienced by pre-menopausal and menopausal women. Methods: This qualitative study was conducted over a 2-month period (March 2022 - April 2022) under the auspices of Imperial College Business School, London, UK. Snowball sampling was used to recruit both menopausal (age 45-70) and pre-menopausal participants (age <45), and sampling continued until data saturation was achieved. 16 semi-structured online interviews were conducted, and transcripts were thematically analyzed following Braun and Clarke’s six-step methodology. Results: A total of 7 higher themes regarding safe discussion of menopausal symptom management were identified by both pre-menopausal and menopausal women, including: “ineffective coping mechanisms”, “perceived onus to self-endure”, “lack of workplace support”, “poor knowledge of management approaches”, “poor healthcare infrastructure”, “poor support from friends and family”, “lack of knowledge and interest from a young age”. Conclusion: Identifying the barriers regarding safe discussion helped to highlight which areas require most significant intervention. Alongside tackling the barriers, menopausal women face, ultimately, there is a pertinent need to and address the lack of insight into menopause from a younger age and to encourage earlier discussions so as to not propagate the cycle of stigma.

Keywords: menopause, stigma, safe discussions, symptom management

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Authors: Tiago Sigal Linhares, Ana Amélia Nascimento da Silva Bones, Juliana Miola, McArthur Alexander Barrow, Airton Tetelbom Stein

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Introduction: As a result of increased globalization and changing migration trends, it is expected that a significant portion of People Living with HIV/AIDS (PLWHA) will change their place of residence over time. In order to provide better health care, monitor the HIV epidemic and plan urban public health care and policies, there is a growing need to formulate a strategy for monitoring PLWHA care, location and migration patterns. The Porto Alegre District is characterized by a high prevalence of PLWHA and is considered one of the epicenters of HIV epidemic in Latin America. Objectives: The aim of this study is to create a digital and easily editable map in order to create a visual representation of the location of PLWHA and to monitor their migration within the city and the country in an effort to promote longitudinal care. Methods: This Experience Report used Google Maps Map Creator to generate an active digital map showing the location and changes in residence of 165 PLWHA who received care at two Primary Health Care (PHC) clinics, which attended an estimated population of five thousand patients, in downtown Porto Alegre over the last four years. Their current addresses were discovered in the unified Brazilian health care system digital records (e-SUS) and updated on the map. Results: A digital map with PLWHA current residence location was created. It was possible to demonstrate visually areas with a large concentration of PLWHA and the migration of the population within the city as wells as other cities, regions and states. Conclusions: An easily reproducible and free map could aid in PLWHA monitoring, urban public health planning, target interventions and situational diagnosis. Moreover, a visual representation of PLWHA location and migration could help bring more attention and investments to areas with geographic inequities or higher prevalence of PLWHA. It also enables notification of local PHC units of monitored patients inside their area, which are in clinical risk or with treatment abandonment through active case findings, improving the care of PLWHA.

Keywords: health care, medical public health, theoretical and conceptual innovations, urban public health

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Authors: A. Aljohani, D. Burke, D. Clarke, M. Gormally, M. Byrne, G. Fleming

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Introduction: House Dust Mites (HDMs) are a source of allergen particles embedded in textiles and furnishings. Vacuum sampling is commonly used to recover and determine the abundance of HDMs but the efficiency of this method is less than standardized. Here, the efficiency of recovery of HDMs was evaluated from home-associated textiles using vacuum sampling protocols.Methods/Approach: Living Mites (LMs) or dead Mites (DMs) House Dust Mites (Dermatophagoides pteronyssinus: FERA, UK) were separately seeded onto the surfaces of Smooth Cotton, Denim and Fleece (25 mites/10x10cm2 squares) and left for 10 minutes before vacuuming. Fabrics were vacuumed (SKC Flite 2 pump) at a flow rate of 14 L/min for 60, 90 or 120 seconds and the number of mites retained by the filter (0.4μm x 37mm) unit was determined. Vacuuming was carried out in a linear direction (Protocol 1) or in a multidirectional pattern (Protocol 2). Additional fabrics with LMs were also frozen and then thawed, thereby euthanizing live mites (now termed EMs). Results/Findings: While there was significantly greater (p=0.000) recovery of mites (76% greater) in fabrics seeded with DMs than LMs irrespective of vacuuming protocol or fabric type, the efficiency of recovery of DMs (72%-76%) did not vary significantly between fabrics. For fabrics containing EMs, recovery was greatest for Smooth Cotton and Denim (65-73% recovered) and least for Fleece (15% recovered). There was no significant difference (p=0.99) between the recovery of mites across all three mite categories from Smooth Cotton and Denim but significantly fewer (p=0.000) mites were recovered from Fleece. Scanning Electron Microscopy images of HMD-seeded fabrics showed that live mites burrowed deeply into the Fleece weave which reduced their efficiency of recovery by vacuuming. Research Implications: Results presented here have implications for the recovery of HDMs by vacuuming and the choice of fabric to ameliorate HDM-dust sensitization.

Keywords: allergy, asthma, dead, fabric, fleece, live mites, sampling

Procedia PDF Downloads 112

Authors: Karish Thavabalan, Aman Sutaria, Alistair Ovenell, Annabelle Parkhouse, Numan Baydemir, Theodore Lally

Abstract:

Background: Open discussions surrounding menopause are often associated with stigma, with many women feeling uncomfortable engaging in them with friends, colleagues, and healthcare professionals. Though the barriers regarding safe discussions of symptom management experienced by menopausal women are well documented, existing research offers little insight into whether these were shared by pre-menopausal women. This study aimed to explore the barriers regarding safe discussions about menopausal symptom management as perceived or experienced by pre-menopausal and menopausal women. Methods: This qualitative study was conducted over a 2-month period (March 2022 - April 2022) under the auspices of Imperial College Business School, London, UK. Snowball sampling was used to recruit both menopausal (age 45-70) and pre-menopausal participants (age <45) and sampling continued until data saturation was achieved. Sixteen semi-structured online interviews were conducted, and transcripts were thematically analyzed following Braun and Clarke’s six-step methodology. Results: A total of 7 higher themes regarding safe discussion of menopausal symptom management were identified by both pre-menopausal and menopausal women, including: “ineffective coping mechanisms,” “perceived onus to self-endure,” “lack of workplace support,” “poor knowledge of management approaches,” “poor healthcare infrastructure,” “poor support from friends and family,” “lack of knowledge and interest from a young age.” Conclusion: Identifying the barriers regarding safe discussion helped to highlight which areas require the most significant intervention. Alongside tackling the barriers, menopausal women face, ultimately, there is a pertinent need to address the lack of insight into menopause from a younger age and to encourage earlier discussions so as to not propagate the cycle of stigma.

Keywords: menopause, safe discussion, symptom management, stigma

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Authors: Hassen Mohammed, Michelle Clarke, Helen Marshall

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Maternal immunization is an effective strategy to protect pregnant women and their offspring from vaccine-preventable diseases. Despite the recommendation of maternal influenza and more recently pertussis immunization in Australia, uptake of these vaccines has been suboptimal. Monitoring the impact of the current funded vaccine programs for pregnant women is limited. The study aimed to assess the impact of the funded program and determine factors associated with vaccine uptake in pregnant women. This observational prospective study was undertaken between November 2014 and July 2016 at the Women’s and Children’s Hospital in South Australia (WCH). Demographic details and vaccination history from South Australian pregnant women who attended the WCH were reviewed. A standardized self-reported survey was conducted in antenatal care with a follow up telephone interview at 8-10 weeks post-delivery. A midwife delivered immunization program for pregnant women in antenatal clinic commenced in April 2015. Of the 180 pregnant women who completed the survey questionnaire, 75.5% and 80.5 % received maternal influenza and pertussis vaccines respectively. First-time mothers had twice the odds of having received influenza vaccine during pregnancy than multiparous women (OR 2.4; CI 1.14 - 4.94; p= 0.021). The proportion of women who received pertussis vaccine during pregnancy, following the introduction of the midwife delivered pertussis vaccination program (140/155, 90.3%) was significantly higher compared with women who received maternal pertussis vaccination prior to the introduction of the program (5/22, 23.7%, p < 0.001). The odds of women receiving maternal pertussis vaccine following the implementation of the midwife delivered program were 31 times higher than women who delivered babies prior to the program (OR 31.7, CI 10.24- 98.27; p < 0.001). High uptake of influenza and pertussis vaccines during pregnancy can be attained with health care provider recommendation and inclusion of maternal immunization as part of standard antenatal care.

Keywords: influenza, maternal immunization, pertussis, provider recommendation

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Authors: Precious Roselyn Obuobi

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Background: In Ghana, water-borne diseases remain a public health concern due to its impact. While marine pollution has been linked to outbreak of diseases especially in communities along the coast, associated risks such as oil spillage, marine debris, erosion, improper waste disposal and management practices persist. Objective: The study seeks to investigate sanitation practices that contribute to marine pollution in Prampram and the prevalence of selected water-borne diseases (diarrhea and typhoid fever). Method: This study used a descriptive cross-sectional design, employing the mix-method (qualitative and quantitative) approach. Twenty-two (22) participants were selected and semistructured questionnaire were administered to them. Additionally, interviews were conducted to collect more information. Further, an observation check-list was used to aid the data collection process. Secondary data comprising information on water-borne diseases in the district was acquired from the district health directorate to determine the prevalence of selected water-borne diseases in the community. Data Analysis: The qualitative data was analyzed using NVIVO® software by adapting the six steps thematic analysis by Braun and Clarke whiles STATA® version 16 was used to analyze the secondary data collected from the district health directorate. A descriptive statistic employed using mean, standard deviation, frequencies and proportions were used to summarize the results. Results: The results showed that open defecation and indiscriminate waste disposal were the main practices contributing to marine pollution in Prampram and its effect on public health. Conclusion: These findings have implications on public health and the environment, thus effort needs to be stepped up in educating the community on best sanitation practices.

Keywords: environment, sanitation, marine pollution, water-borne diseases

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Authors: Edwin O. Clarke, Akintade O. Adeboyejo

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Intensive use of pesticides resulted in dispersal of pollutants throughout the globe. This study was carried out to investigate persistent Organochlorine pesticides (POPs) in water, sediment and fin fishes, Schilbes mystus and Hemichromis fasciatus from two different sampling stations along River Ogun between the month of June 2012 and January 2013. The Organochlorine pesticides analyzed include DDT (pp’1,1,1-trichloro-2,2-bis-(4-chlorophenyl) ethane), DDD, DDE (pp1,1-dichloro-2, 2-bis-(4-chlorophenyl) ethylene, HCH (gamma 1,2,3,4,5,6-hexachlorocylohexane, HCB hexachlorobenzene),Dieldrin (1,2,3,4,10,10-hexachloro-6,7-epoxy-1,4,4a,5,6,7,8,8a octahydro- 1,4,5,8 dimethanonaphthalene). The analysis was done using Gas Chromatograph with Electron Capture Detector. In water sample, the result showed that PPDDT, Endrin aldehyde, Endrin ketone concentrations were high in both stations. The mean value of Organochlorine analyzed in water range from Beta BHC (0.50±0.10µg/l) to PP DDT (162.86±0.21µg/l) in Kara sample station and Beta BHC (0.20±0.07µg/l) to Endrin Aldehyde (76.47±0.02µg/l) in Odo-Ogun sample station. The levels of POPs obtained in sediments ranged from 0.40±0.23µg/g (Beta BHC) to 259.90 ± 1.00µg/kg (Endosulfan sulfate) in Kara sample station and 0.64±0.00µg/g (Beta BHC) to 379.77 ±0.15 µg/g (Endosulfan sulfate) in Odo-Ogun sample station. The levels of POPs obtained in fin fish samples ranged from 0.29±0.00µg/g (Delta BHC) to 197.87 ± 0.31µg/g (PP DDT) in Kara sample station and in Odo-Ogun sample station the mean value for fish samples range from 0.29 ± 0.00 µg/g (Delta BHC) to 197.87 ± 0.32 µg/g (PP DDT). The study showed that the accumulation of POPs affect the environment and reduce water quality. The results showed that the concentrations were found to exceed the maximum acceptable concentration of 0.10µg/l value set by the European Union for the protection of freshwater aquatic life and this can be hazardous if the trend is not checked.

Keywords: hazardous, persistent, pesticides, biomes

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Authors: Amelia Faotusia

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The blue economy concept was coined by Pacific nations in recognition of the importance of sustainably managing their extensive marine territories. This is especially important for major ocean-based economic sectors of Pacific economies, such as coastal tourism. There is an absence of research, however, on the key ways in which the blue economy concept has emerged in discourse and public policy in Pacific countries, as well as how it articulates with coastal tourism. This research helps to fill such a gap with a specific focus on Tonga through the application of a post-positivist research approach to conduct a desktop study of relevant national documents and qualitative interviews with relevant government staff, civil society organizations, and tourism operators. The findings of the research reflect the importance of institutional integration and partnerships for a successful blue economy transition and are presented in the form of two case studies corresponding to two sub-sectors of Tonga’s coastal tourism sector: (i) the whale-watching and swimming industry, and (ii) beach resorts and restaurants. A thematic analysis applied to the interview data of both cases then enabled the identification of key areas and issues for socio-economic policy intervention and recommendations in support of blue economy transitions in Tonga’s coastal tourism sector. Examples of the relevant areas and issues that emerged included the importance of foreign direct investment, local market access, community-based special management areas, as well as the need to address the anthropogenic impacts of tropical cyclones, whale tourism, plastic litter on coastal assets, and ecosystems. Policy and practical interventions in support of addressing such issues include a proposed restructuring of the whale-watching and swimming licensing system; integration of climate resilience, adaptation, and capacity building as priorities of local blue economy interventions; as well as strengthening of the economic sustainability dimension of blue economy policies. Finally, this research also revealed the need for further specificity and research on the influence and value of local Tongan culture and traditional knowledge, particularly within existing customary marine tenure systems, on Tonga’s national and sectoral blue economy policies and transitions.

Keywords: blue economy, coastal tourism, integrated ocean management, ecosystem resilience

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Authors: Naomi Huang Wenya, Xin Xiaohui, Vijaya Rao, Wong Ting Hway, Chow Kah Hoe Pierce, Tan Hiang Khoon

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Background: The majority of the cancelled elective surgeries caused by the COVID-19 pandemic globally were estimated to occur in low- and middle-income countries (LMICs), where surgical services had long been in short supply even before the pandemic. Therefore, minimising disruption to existing surgical care in LMICs is of crucial importance during a pandemic. This study aimed to explore contributory factors to the continuity of surgical care in LMICs, in the face of a pandemic. Methods: Semi-structured interviews were conducted over zoom, with surgical leaders of 25 tertiary hospitals from 11 LMICs in South and Southeast Asia, from September to October 2020. Key themes were subsequently identified from the interview transcripts, using Braun and Clarke's method of thematic analysis. Results: The COVID-19 pandemic affected all surgical services of participating institutions but to varying degrees. Overall, elective surgeries suffered the gravest disruption, followed by outpatient surgical care, and finally, emergency surgeries. Keeping healthcare workers safe and striving for continuity of essential surgical care emerged as notable response strategies observed across all participating institutions. Conclusion: This study suggested that four factors are important for the resilience of surgical care against COVID-19: adequate COVID-19 testing capacity and effective institutional infection control measures, designated COVID-19 treatment facilities, a whole-system approach to balancing pandemic response and meeting essential surgical needs, and active community engagement. These findings can inform healthcare institutions in other countries, especially LMICs, in their effort to tread a fine line between preserving healthcare capacity for pandemic response and protecting surgical services against pandemic disruption.

Keywords: COVID-19, pandemic, LMICs, continuity of surgical service

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Authors: Tria Moore Aimee Walker-Clarke

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The purpose of the study was to develop a deeper understanding of how self-identity is negotiated and reconstructed by people in recovery from substance abuse. The approach draws on the notion that self-identity is constructed through stories. Specifically, dominant narratives of substance abuse involve the 'addict identity' in which the meaning of being an addict is constructed though social interaction and informed by broader social meanings of substance misuse, which are considered deviant. The addict is typically understood as out of control, weak and feckless. Users may unconsciously embody this addict identity which makes recovery less likely. Typical approaches to treatment employ the notion that recovery is much more likely when users change the way they think and feel about themselves by assembling a new identity. Recovery, therefore, involves a reconstruction of the self in a new light, which may mean rejecting a part of the self (the addict identity). One limitation is that previous research on this topic has been quantitative which, while useful, tells us little about how this process is best managed. Should one, for example, reject the past addict identity completely and move on to the new identity, or, is it more effective to accept the past identity and use this in the formation of the new non-user identity? The purpose of this research, then, is to explore how addicts in recovery have managed the transition between their past and current selves and whether this may inform therapeutic practice. Using a narrative approach, data were analyzed from five in-depth interviews with former addicts who had been abstinent for at least a year, and who were in some form of volunteering role at substance treatment services in the UK. Although participants' identified with a previous ‘addict identity,’ and made efforts to disassociate themselves from this, they also recognized that acceptance was an important part of reconstructing their new identity. The participants' narratives used familiar plot lines to structure their stories, in which they positioned themselves as the heroes in their own stories, rather than as victim of circumstance. Instead of rejecting their former addict identity, which would mean rejecting a part of the self, participants used their experience in a reconstructive and restorative way. The findings suggest that encouraging people to tell their story and accept their addict identity are important factors in successful recovery.

Keywords: addiction, identity, narrative, recovery, substance abuse

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Authors: Jason Clarke, Michaela Porubanova, Angela Mazzoli, Gulsah Kut

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What accounts for our perception that time inexorably passes in one direction, from the past to the future, the so-called arrow of time, given that the laws of physics permit motion in one temporal direction to also happen in the reverse temporal direction? Modern physics says that the reason for time’s unidirectional physical arrow is the relationship between time and entropy, the degree of disorder in the universe, which is evolving from low entropy (high order; thermal disequilibrium) toward high entropy (high disorder; thermal equilibrium), the second law of thermodynamics. Accordingly, our perception of the direction of time, from past to future, is believed to emanate as a result of the natural evolution of entropy from low to high, with low entropy defining our notion of ‘before’ and high entropy defining our notion of ‘after’. Here we explored this proposed relationship between entropy and the perception of time’s arrow. We predicted that if the brain has some mechanism for detecting entropy, whose output feeds into processes involved in constructing our perception of the direction of time, presentation of violations to the expectation that low entropy defines ‘before’ and high entropy defines ‘after’ would alert this mechanism, leading to measurable behavioral effects, namely a disruption in duration perception. To test this hypothesis, participants were shown briefly-presented (1000 ms or 500 ms) computer-generated visual dynamic events: novel 3D shapes that were seen either to evolve from whole figures into parts (low to high entropy condition) or were seen in the reverse direction: parts that coalesced into whole figures (high to low entropy condition). On each trial, participants were instructed to reproduce the duration of their visual experience of the stimulus by pressing and releasing the space bar. To ensure that attention was being deployed to the stimuli, a secondary task was to report the direction of the visual event (forward or reverse motion). Participants completed 60 trials. As predicted, we found that duration reproduction was significantly longer for the high to low entropy condition compared to the low to high entropy condition (p=.03). This preliminary data suggests the presence of a neural mechanism that detects entropy, which is used by other processes to construct our perception of the direction of time or time’s arrow.

Keywords: time perception, entropy, temporal illusions, duration perception

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Authors: Kiatkulchai Jitt-Aer, Graham Wall, Dylan Jones

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As a result of the Indian Ocean tsunami in 2004, logistics applied to disaster relief operations has received great attention in the humanitarian sector. As learned from such disaster, preparing and responding to the aspect of delivering essential items from distribution centres to affected locations are of the importance for relief operations as the nature of disasters is uncertain especially in suffering figures, which are normally proportional to quantity of supplies. Thus, this study proposes a spatial decision support system (SDSS) for humanitarian logistics by integrating Geographical Information Systems (GIS) and the capacitated vehicle routing problem (CVRP). The GIS is utilised for acquiring demands simulated from the tsunami flooding model of the affected area in the first stage, and visualising the simulation solutions in the last stage. While CVRP in this study encompasses designing the relief routes of a set of homogeneous vehicles from a relief centre to a set of geographically distributed evacuation points in which their demands are estimated by using both simulation and randomisation techniques. The CVRP is modeled as a multi-objective optimization problem where both total travelling distance and total transport resources used are minimized, while demand-cost efficiency of each route is maximized in order to determine route priority. As the model is a NP-hard combinatorial optimization problem, the Clarke and Wright Saving heuristics is proposed to solve the problem for the near-optimal solutions. The real-case instances in the coastal area of Phuket, Thailand are studied to perform the SDSS that allows a decision maker to visually analyse the simulation scenarios through different decision factors.

Keywords: demand simulation, humanitarian logistics, geographical information systems, relief operations, capacitated vehicle routing problem

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Authors: Muhammad Bilal, Amelia Jane Llyod, Habsah Mohamad, Jia Hui Wong, Abdul Aziz Mohamed Yusoff, Jafri Malin Abdullah, Jingli Zhang

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Epilepsy is one of the major brain afflictions occurs with uncontrolled excitation of cortex; disturbed 50 million of world’s population. About 25 percent of patients subjected to adverse effects from antiepileptic drugs (AEDs) such as depression, nausea, tremors, gastrointestinal symptoms, osteoporosis, dizziness, weight change, drowsiness, fatigue are commonly observed indications; therefore, new drugs are required to cure epilepsy. GABA is principle inhibitory neurotransmitter, control excitation of the brain. Mutation or dysfunction of GABA receptor is one of the primary causes of epilepsy, which is confirmed from many acquired models of epilepsy like traumatic brain injury, kindling, and status epilepticus models of epilepsy. GABA receptor has 3 distinct types such as GABA (A), GABA (B), GABA(C).GABA (A) receptor has 20 different subunits, α1β2γ2 subunits composition of GABA (A) receptor is the most used combination of subunits for screening of compounds against epilepsy. We expressed α1β2γ2s subunits of GABA (A) Receptor in Xenopus leavis oocytes and examined the enhancement potential of 4-Hydroxybenzoic acid compound on GABA (A) receptor via two-electrode voltage clamp current recording technique. Bamboo shoots are the young, tender offspring of bamboo, which are usually harvested after a cultivating period of 2 weeks. Proteins, acids, fat, starch, carbohydrate, fatty acid, vitamin, dietary fiber, and minerals are the major constituent found systematically in bamboo shoots. These shoots reported to have anticancer, antiviral, antibacterial activity, also possess antioxidant properties due to the presence of phenolic compounds. Student t-test analysis suggested that 4- hydroxybenzoic acid positively allosteric GABA (A) receptor, increased normalized current amplitude to 1.0304±0.0464(p value 0.032) compared with vehicle. 4-Hydrobenzoic acid, a compound from Dendrocalamus Asper bamboo shoot gives new insights for future studies on bamboo shoots with motivation for extraction of more compounds to investigate their effects on human and rodents against epilepsy, insomnia, and anxiety.

Keywords: α1β2γ2S, antiepileptic, bamboo shoots, epilepsy GABA (A) receptor, two-microelectrode voltage clamp, xenopus laevis oocytes

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Authors: Amelia Frickey, Timothy (TJ) Sheridan, Angelica Rossi, Bahar Aliakbarian

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The growth of large food supply chains demanded improved end-to-end traceability of food products, which has led to companies being increasingly interested in using smart technologies such as Radio Frequency Identification (RFID)-enabled packaging to track items. As technology is being widely used, there are several technological or economic issues that should be overcome to facilitate the adoption of this track-and-trace technology. One of the technological challenges of RFID technology is its sensitivity to different environmental form factors, including packaging materials and the content of the packaging. Although researchers have assessed the performance loss due to the proximity of water and aqueous solutions, there is still the need to further investigate the impacts of food products on the reading range of RFID tags. However, to the best of our knowledge, there are not enough studies to determine the correlation between RFID tag performance and food beverages properties. The goal of this project was to investigate the effect of the solution properties (pH and conductivity) and different packaging materials filled with food-like water-based solutions on the performance of an RFID tag. Three commercially available ultra high-frequency RFID tags were placed on three different bottles and filled with different concentrations of water-based solutions, including sodium chloride, citric acid, sucrose, and ethanol. Transparent glass, Polyethylneterephtalate (PET), and Tetrapak® were used as the packaging materials commonly used in the beverage industries. Tag readability (Theoretical Read Range, TRR) and sensitivity (Power on Tag Forward, PoF) were determined using an anechoic chamber. First, the best place to attach the tag for each packaging material was investigated using empty and water-filled bottles. Then, the bottles were filled with the food-like solutions and tested with the three different tags and the PoF and TRR at the fixed frequency of 915MHz. In parallel, the pH and conductivity of solutions were measured. The best-performing tag was then selected to test the bottles filled with wine, orange, and apple juice. Despite various solutions altering the performance of each tag, the change in tag performance had no correlation with the pH or conductivity of the solution. Additionally, packaging material played a significant role in tag performance. Each tag tested performed optimally under different conditions. This study is the first part of comprehensive research to determine the regression model for the prediction of tag performance behavior based on the packaging material and the content. More investigations, including more tags and food products, are needed to be able to develop a robust regression model. The results of this study can be used by RFID tag manufacturers to design suitable tags for specific products with similar properties.

Keywords: smart food packaging, supply chain management, food waste, radio frequency identification

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Authors: I. Tzoka, V. A. Chirayath, A. Brandt, J. Asaadi, Melvin J. Aviles, Stephen Clarke, Stefan Cwik, Michael R. Foley, Cole J. Hamel, Alexey Lyashenko, Michael J. Minot, Mark A. Popecki, Michael E. Stochaj, S. Shin

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Micro-channel plate photo-multiplier tubes (MCP-PMTs) have become ubiquitous and are widely considered potential candidates for next generation High Energy Physics experiments due to their picosecond timing resolution, ability to operate in strong magnetic fields, and low noise rates. A key factor that determines the applicability of MCP-PMTs in their lifetime, especially when they are used in high event rate experiments. We have developed a novel method for the investigation of the aging behavior of an MCP-PMT on an accelerated basis. The method involves exposing a localized region of the MCP-PMT to photons at a high repetition rate. This pixel-based method was inspired by earlier results showing that damage to the photocathode of the MCP-PMT occurs primarily at the site of light exposure and that the surrounding region undergoes minimal damage. One advantage of the pixel-based method is that it allows the dynamics of photo-cathode damage to be studied at multiple locations within the same MCP-PMT under different operating conditions. In this work, we use the pixel-based accelerated lifetime test to investigate the aging behavior of a 20 cm x 20 cm Large Area Picosecond Photo Detector (LAPPD) manufactured by INCOM Inc. at multiple locations within the same device under different operating conditions. We compare the aging behavior of the MCP-PMT obtained from the first lifetime test conducted under high gain conditions to the lifetime obtained at a different gain. Through this work, we aim to correlate the lifetime of the MCP-PMT and the rate of ion feedback, which is a function of the gain of each MCP, and which can also vary from point to point across a large area (400 $cm^2$) MCP. The tests were made possible by the uniqueness of the LAPPD design, which allows independent control of the gain of the chevron stacked MCPs. We will further discuss the implications of our results for optimizing the operating conditions of the detector when used in high event rate experiments.

Keywords: electron multipliers (vacuum), LAPPD, lifetime, micro-channel plate photo-multipliers tubes, photoemission, time-of-flight

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Authors: Mamochana, A. Ramatea, Fumane, P. Khanare

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Enabling the learning environment is a significant and adaptive technique necessary to navigate learners’ educational challenges. However, research has indicated that quality provision of education in the environments that are enabling, especially to learners with visual impairments (LVIs, hereafter) in rural schools, remain an ongoing challenge globally. Hence, LVIs often have a lower level of academic performance as compared to their peers. To balance this gap and fulfill learners'fundamentalhuman rights¬ of receiving an equal quality education, appropriate measures and structures that make enabling learning environment a better place to learn must be better understood. This paper, therefore, intends to find possible means that rural schools of Lesotho can employ to make the learning environment for LVIs enabling. The present study aims to determine suitable assets that can be drawn to make the learning environment for LVIs enabling. The study is also informed by the transformative paradigm and situated within a qualitative research approach. Data were generated through focus group discussions with twelve teachers who were purposefully selected from two rural primary schools in Lesotho. The generated data were then analyzed thematically using Braun and Clarke's six-phase framework. The findings of the study indicated that participating teachers do have an understanding that rural schools boast of assets (existing and hidden) that have a positive influence in responding to the special educational needs of LVIs. However, the participants also admitted that although their schools boast of assets, they still experience limited knowledge about the use of the existing assets and thus, realized a need for improved collaboration, involvement of the existing assets, and enhancement of academic resources to make LVIs’ learning environment enabling. The findings of this study highlight the significance of the effective use of assets. Additionally, coincides with literature that shows recognizing and tapping into the existing assets enable learning for LVIs. In conclusion, the participants in the current study indicated that for LVIs’ learning environment to be enabling, there has to be sufficient use of the existing assets. The researchers, therefore, recommend that the appropriate use of assets is good, but may not be sufficient if the existing assets are not adequately managed. Hence,VILs experience a vicious cycle of vulnerability. It was thus, recommended that adequate use of assets and teachers' engagement as active assets should always be considered to make the learning environment a better place for LVIs to learan in the future

Keywords: assets, enabling learning environment, rural schools, learners with visual impairments

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Authors: Ali Alshumrani, Nathan Clarke, Bogdan Ghite, Stavros Shiaeles

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Digital forensics has become an essential tool in the investigation of cyber and computer-assisted crime. Arguably, given the prevalence of technology and the subsequent digital footprints that exist, it could have a significant role across almost all crimes. However, the variety of technology platforms (such as computers, mobiles, Closed-Circuit Television (CCTV), Internet of Things (IoT), databases, drones, cloud computing services), heterogeneity and volume of data, forensic tool capability, and the investigative cost make investigations both technically challenging and prohibitively expensive. Forensic tools also tend to be siloed into specific technologies, e.g., File System Forensic Analysis Tools (FS-FAT) and Network Forensic Analysis Tools (N-FAT), and a good deal of data sources has little to no specialist forensic tools. Increasingly it also becomes essential to compare and correlate evidence across data sources and to do so in an efficient and effective manner enabling an investigator to answer high-level questions of the data in a timely manner without having to trawl through data and perform the correlation manually. This paper proposes a Unified Forensic Analysis Tool (U-FAT), which aims to establish a common language for electronic information and permit multi-source forensic analysis. Core to this approach is the identification and development of forensic analyses that automate complex data correlations, enabling investigators to investigate cases more efficiently. The paper presents a systematic analysis of major crime categories and identifies what forensic analyses could be used. For example, in a child abduction, an investigation team might have evidence from a range of sources including computing devices (mobile phone, PC), CCTV (potentially a large number), ISP records, and mobile network cell tower data, in addition to third party databases such as the National Sex Offender registry and tax records, with the desire to auto-correlate and across sources and visualize in a cognitively effective manner. U-FAT provides a holistic, flexible, and extensible approach to providing digital forensics in technology, application, and data-agnostic manner, providing powerful and automated forensic analysis.

Keywords: digital forensics, evidence correlation, heterogeneous data, forensics tool

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Authors: K. Al Ammari, B. G. Clarke

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Soil improvement using vibro stone column techniques consists of two main parts: (1) the installed load bearing columns of well-compacted, coarse-grained material and (2) the improvements to the surrounding soil due to vibro compaction. Extensive research work has been carried out over the last 20 years to understand the improvement in the composite foundation performance due to the second part mentioned above. Nevertheless, few of these studies have tried to quantify some of the key design parameters, namely the changes in the stiffness and stress state of the treated soil, or have consider these parameters in the design and calculation process. Consequently, empirical and conservative design methods are still being used by ground improvement companies with a significant variety of results in engineering practice. Two-dimensional finite element study to develop an axisymmetric model of a single stone column reinforced foundation was performed using PLAXIS 2D AE to quantify the effect of the vibro installation of this column in soft saturated clay. Settlement and bearing performance were studied as an essential part of the design and calculation of the stone column foundation. Particular attention was paid to the large deformation in the soft clay around the installed column caused by the lateral expansion. So updated mesh advanced option was taken in the analysis. In this analysis, different degrees of stone column lateral expansions were simulated and numerically analyzed, and then the changes in the stress state, stiffness, settlement performance and bearing capacity were quantified. It was found that application of radial expansion will produce a horizontal stress in the soft clay mass that gradually decrease as the distance from the stone column axis increases. The excess pore pressure due to the undrained conditions starts to dissipate immediately after finishing the column installation, allowing the horizontal stress to relax. Changes in the coefficient of the lateral earth pressure K ٭, which is very important in representing the stress state, and the new stiffness distribution in the reinforced clay mass, were estimated. More encouraging results showed that increasing the expansion during column installation has a noticeable effect on improving the bearing capacity and reducing the settlement of reinforced ground, So, a design method should include this significant effect of the applied lateral displacement during the stone column instillation in simulation and numerical analysis design.

Keywords: bearing capacity, design, installation, numerical analysis, settlement, stone column

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Authors: J. B. Athavale, Sean Clarke

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Introduction According to ancient Indian scriptures, a person’s consciousness includes the physical body, the vital energy sheath (Pranshakti), the mental body (which includes one’s feelings and emotions), the intellectual body (which refers to one’s decision-making ability), and the Soul (which is the God Principle that resides in every person). Apart from the physical body, all the other aspects are subtle in nature. In today’s world, much attention is given to one’s physical appearance and intellectual prowess. While there have been improvements in the attention given to mental health, its complete nature is not understood, and in many cultures, mental ill health is considered taboo and looked down upon. Regarding the spiritual well-being of a person, our spiritual research has shown that people’s understanding and efforts are mostly lacking and superficial as they do not conform to Universal Spiritual Principles. Also, true well-being occurs only when all the bodies are healthy. Methodology The spiritual research team at the University has found that the spiritual aspect of a person’s life affects all the physical, psychological, and intellectual bodies of a person resulting in ill health. Cleansing these bodies at a spiritual level is essential to regain well-being. Using Aura and Energy Scanners and advanced sixth sense, we studied what causes spiritual impurity in various bodies and how to cleanse them. We measured the spiritual vibrations of a person and how they get affected due to various daily activities. For example, we studied the difference in a person’s aura before and after applying chemical-based makeup vs. natural makeup. Key Findings From the various spiritual research experiments we conducted, we found that: • All our actions and our thoughts affect our various bodies and have the potential to change the aura for the better or worse. • When there is an increase in negative vibrations around a person, negative energies from the subtle dimension are more likely to affect a person. • As the person’s spiritual level increases, the positivity in their aura also increases, and it is much easier to cleanse the various bodies spiritually. • Spiritual practice is like a general spiritual tonic that increases the positivity in one’s aura. The benefits of this are that it leads to mental stability and intellectual clarity. • Spiritual healing remedies augment any spiritual practice to obtain a faster healing effect. Conclusion Taking care of oneself spiritually has a positive halo effect on all one’s bodies. Spiritual cleansing is required regularly if one wants to attain a state of well-being. Spiritual practice and spiritual healing lead to spiritual growth, stability of mind, and less stress and reactions. Spiritually purer people affect the environment positively, and there is less unrest and more harmony between man and nature.

Keywords: body, spirituality, cleansing, consciousness

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Authors: Germans Natuhwera, Peter Ellis, Acuda Wilson, Anne Merriman, Martha Rabwoni

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Objective: The aim of the study was to diagnose the socio-economic burden and impact of a diagnosis of cervical cancer (CC) in rural women in the context of low-resourced country Uganda, using a phenomenological enquiry. Methods: This was a multi-site phenomenological inquiry, conducted at three hospice settings; Mobile Hospice Mbarara in southwestern, Little Hospice Hoima in Western, and Hospice Africa Uganda Kampala in central Uganda. A purposive sample of women with a histologically confirmed diagnosis of CC was recruited. Data was collected using open-ended audio-recorded interviews conducted in the native languages of participants. Interviews were transcribed verbatim in English, and Braun and Clarke’s (2019) framework of thematic analysis was used. Results: 13 women with a mean age of 49.2 and age range 29-71 participated in the study. All participants were of low socioeconomic status. The majority (84.6%) had advanced disease at diagnosis. A fuller reading of transcripts produced four major themes clustered under; (1) socioeconomic characteristics of women, (2) impact of CC on women’s relationships, (3) disrupted and impaired activities of daily living (ADLs), and (4) economic disruptions. Conclusions: A diagnosis of CC introduces significant socio-economic disruptions in a woman’s and her family’s life. CC causes disability, impairs the woman and her family’s productivity hence exacerbating levels of poverty in the home. High and expensive out-of-pocket expenditure on treatment, investigations, and transport costs further compound the socio-economic burden. Decentralizing cancer care services to regional centers, scaling up screening services, subsidizing costs of cancer care services, or making cervical cancer care treatment free of charge, strengthening monitoring mechanisms in public facilities to curb the vice of healthcare workers soliciting bribes from patients, increased mass awareness campaigns about cancer, training more healthcare professionals in cancer investigation and management, and palliative care, and introducing an introductory course on gynecologic cancers into all health training institutions are recommended.

Keywords: activities of daily living, cervical cancer, out-of-pocket, expenditure, phenomenology, socioeconomic

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Authors: Vivek Rangarajan, Kim G. Clarke

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Bacillus lipopeptides are biosurfactants with wide ranging applications in the medical, food, agricultural, environmental and cosmetic industries. They are produced as a mix of three families, surfactin, iturin and fengycin, each comprising a large number of homologues of varying functionalities. Consequently, the method and degree of purification of the lipopeptide cocktail becomes particularly important if the functionality of the lipopeptide end-product is to be maximized for the specific application. However, downstream processing of Bacillus lipopeptides is particularly challenging due to the subtle variations observed in the different lipopeptide homologues and isoforms. To date, the most frequently used lipopeptide purification operations have been acid precipitation, solvent extraction, membrane ultrafiltration, adsorption and size exclusion. RP-HPLC (reverse phase high pressure liquid chromatography) also has potential for fractionation of the lipopeptide homologues. In the studies presented here, membrane ultrafiltration and RP-HPLC were evaluated for lipopeptide purification to different degrees of purities for maximum functionality. Batch membrane ultrafiltration using 50 kDa polyether sulphone (PES) membranes resulted in lipopeptide recovery of about 68% for surfactin and 82 % for fengycin. The recovery was further improved to 95% by using size-conditioned lipopeptide micelles. The conditioning of lipopeptides with Ca2+ ions resulted in uniformly sized micelles with average size of 96.4 nm and a polydispersity index of 0.18. The size conditioning also facilitated removal of impurities (molecular weight ranging between 2335-3500 Da) through operation of the system under dia-filtration mode, in a way similar to salt removal from protein by dialysis. The resultant purified lipopeptide was devoid of macromolecular impurities and could ideally suit applications in the cosmetic and food industries. Enhanced purification using RP-HPLC was carried out in an analytical C18 column, with the aim to fractionate lipopeptides into their constituent homologues. The column was eluted with mobile phase comprising acetonitrile and water over an acetonitrile gradient, 35% - 80%, over 70 minutes. The gradient elution program resulted in as many as 41 fractions of individual lipopeptide homologues. The efficacy test of these fractions against fungal phytopathogens showed that first 21 fractions, identified to be homologues of iturins and fengycins, displayed maximum antifungal activities, suitable for biocontrol in the agricultural industry. Thus, in the current study, the downstream processing of lipopeptides leading to tailor-made products for selective applications was demonstrated using two major downstream unit operations.

Keywords: bacillus lipopeptides, membrane ultrafiltration, purification, RP-HPLC

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Authors: Amelia J. McFarland, Andrew K. Davey, Shailendra Anoopkumar-Dukie

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Microglia are the resident macrophage population of the central nervous system (CNS), contributing to both innate and adaptive immune response, and brain homeostasis. Activation of microglia occurs in response to a multitude of pathogenic stimuli in their microenvironment; this induces morphological and functional changes, resulting in a state of acute neuroinflammation which facilitates injury resolution. Adequate microglial function is essential for the health of the neuroparenchyma, with microglial dysfunction implicated in numerous CNS pathologies. Given the critical role that these macrophage-derived cells play in CNS homeostasis, there is a high demand for microglial models suitable for use in neuroscience research. The isolation of primary human microglia, however, is both difficult and costly, with microglial activation an unwanted but inevitable result of the extraction process. Consequently, there is a need for the development of alternative experimental models which exhibit morphological, biochemical and functional characteristics of human microglia without the difficulties associated with primary cell lines. In this study, our aim was to evaluate whether THP-1 human peripheral blood monocytes would display microglial-like qualities following an induced differentiation, and, therefore, be suitable for use as surrogate microglia. To achieve this aim, THP-1 human peripheral blood monocytes from acute monocytic leukaemia were differentiated with a range of phorbol 12-myristate 13-acetate (PMA) concentrations (50-200 nM) using two different protocols: a 5-day continuous PMA exposure or a 3-day continuous PMA exposure followed by a 5-day rest in normal media. In each protocol and at each PMA concentration, microglial-like cell morphology was assessed through crystal violet staining and the presence of CD-14 microglial / macrophage cell surface marker. Lipopolysaccharide (LPS) from Escherichia coli (055: B5) was then added at a range of concentrations from 0-10 mcg/mL to activate the PMA-differentiated THP-1 cells. Functional microglial-like behavior was evaluated by quantifying the release of prostaglandin (PG)-E2 and pro-inflammatory cytokines interleukin (IL)-1β and tumour necrosis factor (TNF)-α using mediator-specific ELISAs. Furthermore, production of global reactive oxygen species (ROS) and nitric oxide (NO) were determined fluorometrically using dichlorodihydrofluorescein diacetate (DCFH-DA) and diaminofluorescein diacetate (DAF-2-DA) respectively. Following PMA-treatment, it was observed both differentiation protocols resulted in cells displaying distinct microglial morphology from 10 nM PMA. Activation of differentiated cells using LPS significantly augmented IL-1β, TNF-α and PGE2 release at all LPS concentrations under both differentiation protocols. Similarly, a significant increase in DCFH-DA and DAF-2-DA fluorescence was observed, indicative of increases in ROS and NO production. For all endpoints, the 5-day continuous PMA treatment protocol yielded significantly higher mediator levels than the 3-day treatment and 5-day rest protocol. Our data, therefore, suggests that the differentiation of THP-1 human monocyte cells with PMA yields a homogenous microglial-like population which, following stimulation with LPS, undergo activation to release a range of pro-inflammatory mediators associated with microglial activation. Thus, the use of PMA-differentiated THP-1 cells represents a suitable microglial model for in vitro research.

Keywords: differentiation, lipopolysaccharide, microglia, monocyte, neuroscience, THP-1

Procedia PDF Downloads 355