Search results for: supervisory control and data acquisition system

Authors: Rachael Andrews, Viktoria McMillan, Shuaib Nasser, Christopher M. Roberts

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Background and objective: The National Review of Asthma Deaths (NRAD) found that asthma management and care was inadequate in 26% of cases reviewed. Major shortfalls identified were adherence to national guidelines and standards and, particularly, the organisation of care, including supervision and monitoring in primary care, with 70% of cases reviewed having at least one avoidable factor in this area. 5.4 million people in the UK are diagnosed with and actively treated for asthma, and approximately 60,000 are admitted to hospital with acute exacerbations each year. The majority of people with asthma receive management and treatment solely in primary care. This has therefore created concern that many people within the UK are receiving sub-optimal asthma care resulting in unnecessary morbidity and risk of adverse outcome. NRAD concluded that a national asthma audit programme should be established to measure and improve processes, organisation, and outcomes of asthma care. Objective: To develop a primary care dataset enabling extraction of information from GP practices in Wales and providing robust data by which results and lessons could be drawn and drive service development and improvement. Methods: A multidisciplinary group of experts, including general practitioners, primary care organisation representatives, and asthma patients was formed and used as a source of governance and guidance. A review of asthma literature, guidance, and standards took place and was used to identify areas of asthma care which, if improved, would lead to better patient outcomes. Modified Delphi methodology was used to gain consensus from the expert group on which of the areas identified were to be prioritised, and an asthma patient and carer focus group held to seek views and feedback on areas of asthma care that were important to them. Areas of asthma care identified by both groups were mapped to asthma guidelines and standards to inform and develop primary and secondary care datasets covering both adult and pediatric care. Dataset development consisted of expert review and a targeted consultation process in order to seek broad stakeholder views and feedback. Results: Areas of asthma care identified as requiring prioritisation by the National Asthma Audit were: (i) Prescribing, (ii) Asthma diagnosis (iii) Asthma Reviews (iv) Personalised Asthma Action Plans (PAAPs) (v) Primary care follow-up after discharge from hospital (vi) Methodologies and primary care queries were developed to cover each of the areas of poor and variable asthma care identified and the queries designed to extract information directly from electronic patients’ records. Conclusion: This paper describes the methodological approach followed to develop primary care datasets for a National Asthma Audit. It sets out the principles behind the establishment of a National Asthma Audit programme in response to a national asthma mortality review and describes the development activities undertaken. Key process elements included: (i) mapping identified areas of poor and variable asthma care to national guidelines and standards, (ii) early engagement of experts, including clinicians and patients in the process, and (iii) targeted consultation of the queries to provide further insight into measures that were collectable, reproducible and relevant.

Keywords: asthma, primary care, general practice, dataset development

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Authors: Bonnie R. Brammall, Rhonda M. Garad, Helena J. Teede, Cheryce L. Harrison

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Introduction: Reproductive aged women are at high-risk for accelerated weight gain and obesity development, with pregnancy recognised as a critical contributory life phase. Healthy lifestyle interventions during the preconception and antenatal period improve maternal and infant health outcomes. Yet, interventions from preconception through to postpartum and translation and implementation into real-world healthcare settings remain limited. OptimalMe is a randomised, hybrid implementation effectiveness study of evidence-based healthy lifestyle intervention. Here, we report engagement, acceptability of the intervention during preconception, and self-reported behaviour change outcomes as a result of the preconception phase of the intervention. Methods: Reproductive aged women who upgraded their private health insurance to include pregnancy and birth cover, signalling a pregnancy intention, were invited to participate. Women received access to an online portal with preconception health and lifestyle modules, goal-setting and behaviour change tools, monthly SMS messages, and two coaching sessions (randomised to video or phone) prior to pregnancy. Results: Overall n=527 expressed interest in participating. Of these, n=33 did not meet inclusion criteria, n=8 were not contactable for eligibility screening, and n=177 failed to engage after the screening, leaving n=309 who were enrolled in OptimalMe and randomised to intervention delivery method. Engagement with coaching sessions dropped by 25% for session two, with no difference between intervention groups. Women had a mean (SD) age of 31.7 (4.3) years and, at baseline, a self-reported mean BMI of 25.7 (6.1) kg/m², with 55.8% (n=172) of a healthy BMI. Behaviour was sub-optimal with infrequent self-weighing (38.1%), alcohol consumption prevalent (57.1%), sub-optimal pre-pregnancy supplementation (61.5%), and incomplete medical screening. Post-intervention 73.2% of women reported engagement with a GP for preconception care and improved lifestyle behaviour (85.5%), since starting OptimalMe. Direct pre-and-post comparison of individual participant data showed that of 322 points of potential change (up-to-date cervical screening, elimination of high-risk behaviours [alcohol, drugs, smoking], uptake of preconception supplements and improved weighing habits) 158 (49.1%) points of change were achieved. Health coaching sessions were found to improve accountability and confidence, yet further personalisation and support were desired. Engagement with video and phone sessions was comparable, having similar impacts on behaviour change, and both methods were well accepted and increased women's accountability. Conclusion: A low-intensity digital health and lifestyle program with embedded health coaching can improve the uptake of preconception care and lead to self-reported behaviour change. This is the first program of its kind to reach an otherwise healthy population of women planning a pregnancy. Women who were otherwise healthy showed divergence from preconception health and lifestyle objectives and benefited from the intervention. OptimalMe shows promising results for population-based behaviour change interventions that can improve preconception lifestyle habits and increase engagement with clinical health care for pregnancy preparation.

Keywords: preconception, pregnancy, preventative health, weight gain prevention, self-management, behaviour change, digital health, telehealth, intervention, women's health

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Authors: Nabila Farhin, Israt Jahan

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Despite the feminisation of the global health force, women mostly engage in nursing, midwifery and community health workers (HWs), and the posts like surgeons, doctors, and specialists are generally male-dominated. It is also prominent in Bangladesh, where female HWs witness systematic workplace inequalities, discrimination, and underpayment. The Covid-19 pandemic put unsurmountable pressure on HWs as they had to serve in high-risk workplaces as frontliners. The already disadvantaged female HWs shouldered the same burden, were overworked without adequate occupational health and safety measures (OSH) and risked their lives. Acknowledging their vulnerable workplace conditions, the World Health Organization (WHO) and International Labour Organization (ILO) circulated a few specialised guidelines amid the peril. Bangladesh tried to adhere to international guidelines while formulating pandemic management strategies. In reality, the already weak and understaffed health sector collapsed with the patient influx and many HWs got infected and died in the line of duty, exposing the high-risk nature of the work. Unfortunately, the gender-segregated data of infected HWs are absent. This qualitative research investigates whether the existing laws of Bangladesh are adequate in protecting female HWs as frontliners in high-risk workplaces during the Covid-19 pandemic. The paper first examines international labour laws safeguarding female frontline HWs. It also analyses the specialised Covid-19 pandemic guidelines protecting their interests. Finally, the research investigates the compliance of Bangladesh as per international legal guidance during the pandemic. In doing so, it explores the domestic laws, professional guidelines for HWs and pandemic response strategies. The paper critically examines the primary sources like international and national statutes, rules, regulations and guidelines. Secondary sources like authoritative journal articles, books and newspaper reports are contextually analysed in line with the objective of the paper. The definition of HW is ambiguous in the labour laws of Bangladesh. It leads to confusion regarding the extent of legal protection rendered to female HWs at private hospitals in high-risk situations. The labour laws are not applicable in Public hospitals, as the employees follow the public service rules. Unfortunately, the country has no specialised law to protect HWs in high-risk workplaces, and the professional guidelines for HWs also remain inadequate in this regard. Even though the pandemic management strategies highlight some protective measures in high-risk situations, they only deal with HWs who are pregnant or have underlying health issues. No specialised protective guidelines can be found for female HWs as frontliners. Therefore, the laws are insufficient and failed to render adequate legal protection to female frontline HWs during the pandemic. The country also lacks comprehensive health legislation and uniform institutional and professional guidelines, preventing them from accessing grievance mechanisms. Hence, the female HWs felt victimised while duty-bound to serve in high-risk workplaces without adequate safeguards. Bangladesh should clarify the definition of HWs and standardise the service rules for providing medical care in high-risk workplaces. The research also recommends adequate health legislation and specialised legal protection to safeguard female HWs in future emergencies.

Keywords: female health workers (HWs), high-risk workplaces, Covid-19 pandemic, Bangladesh

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Authors: Etienne Herrick

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As an economically and ecologically feasible method for farmers to introduce greater diversity into their crop rotations, cover cropping presents a valuable opportunity for improving the sustainability of food production. Planted in-between cash crop growing seasons, cover crops serve to enhance agroecosystem functioning, rather than being destined for sale or consumption. In fact, cover crops may hold the capacity to deliver multiple ecosystem functions or services simultaneously (multifunctionality). Building upon this line of research will not only benefit society at present, but also support its continued survival through its potential for restoring depleted soils and reducing the need for energy-intensive and harmful external inputs like fertilizers and pesticides. This study utilizes a trait-based approach to explore the influence of inter- and intra-specific interactions in summer cover crop mixtures and monocultures on functional trait expression and ecosystem services. Functional traits that enhance ecosystem services related to agricultural production include height, specific leaf area (SLA), root, shoot ratio, leaf C and N concentrations, and flowering phenology. Ecosystem services include biomass production, weed suppression, reduced N leaching, N recycling, and support of pollinators. Employing a trait-based approach may allow for the elucidation of mechanistic links between plant structure and resulting ecosystem service delivery. While relationships between some functional traits and the delivery of particular ecosystem services may be readily apparent through existing ecological knowledge (e.g. height positively correlating with weed suppression), this study will begin to quantify those relationships so as to gain further understanding of whether and how measurable variation in functional trait expression across cover crop mixtures and monocultures can serve as a reliable predictor of variation in the types and abundances of ecosystem services delivered. Six cover crop species, including legume, grass, and broadleaf functional types, were selected for growth in six mixtures and their component monocultures based upon the principle of trait complementarity. The tricultures (three-way mixtures) are comprised of a legume, grass, and broadleaf species, and include cowpea/sudex/buckwheat, sunnhemp/sudex/buckwheat, and chickling vetch/oat/buckwheat combinations; the dicultures contain the same legume and grass combinations as above, without the buckwheat broadleaf. By combining species with expectedly complimentary traits (for example, legumes are N suppliers and grasses are N acquirers, creating a nutrient cycling loop) the cover crop mixtures may elicit a broader range of ecosystem services than that provided by a monoculture, though trade-offs could exist. Collecting functional trait data will enable the investigation of the types of interactions driving these ecosystem service outcomes. It also allows for generalizability across a broader range of species than just those selected for this study, which may aid in informing further research efforts exploring species and ecosystem functioning, as well as on-farm management decisions.

Keywords: agroecology, cover crops, functional traits, multifunctionality, trait complementarity

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Authors: Ado Ismail Sabo

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Statement of the Problem: The global trend in tertiary education institutions today is changing and moving towards engagement, promotion and marketing. The reason is to upscale reputation and impact positioning. More prominently, existing rivalry today seeks to draw-in the best and brightest students. A university or college is no longer just an institution of higher learning, but one adapting additional business nomenclature. Therefore, huge financial resources are invested by educational institutions to polish their image and improve their global and national ranking. In Nigeria, which boasts of a vast population of over 180 million people, some of whose patronage can bolster its education sector; standard of education continues to decline. Today, some Nigerian tertiary education institutions are shadows of their pasts, in terms of academic excellence. Quality has been relinquished because of the unquenchable quest by government officials, some civil servants, school heads and educators to amass wealth. It is very difficult to gain student satisfaction and their loyalty. Some of the student’s loyalties factor towards public higher educational institutions might be confusing. It is difficult to understand the extent to which students are satisfy on many needs. Some students might feel satisfy with the academic lecturers only, whereas others may want everything, and others will never satisfy. Due to these problems, this research aims to uncover the crucial factors influencing student loyalty and to examine if students’ satisfaction might impact mediate the relationship between service quality, organisational image and students’ loyalty towards public higher education institutions in Kano State, Nigeria. The significance of the current study is underscored by the paucity of similar research in the subject area and public tertiary education in a developing country like Nigeria as shown in existing literature. Methodology: The current study was undertaken by quantitative research methodology. Sample of 600 valid responses were obtained within the study population comprising six selected public higher education institutions in Kano State, Nigeria. These include: North West University Kano, Bayero University Kano, School of Management Studies Kano, School of Technology Kano, Sa’adatu Rimi College Kano and Federal College of Education (FCE) Kano. Four main hypotheses were formulated and tested using structural equation modeling techniques with Analysis of Moment Structure (AMOS Version 22.0). Results: Analysis of the data provided support for the main issue of this study, and the following findings are established: “Student Satisfaction mediates the relationship between Service Quality and Student Loyalty”, “Student Satisfaction mediates the relationship between Organizational Image and Student Loyalty” respectively. The findings of this study contributed to the theoretical implication which proposed a structural model that examined the relationships among overall Organizational image, service quality, student satisfaction and student loyalty. Conclusion: In addition, the findings offered a better insight to the managerial (higher institution of learning service providers) by focusing on portraying the image of service quality with student satisfaction in improving the quality of student loyalty.

Keywords: student loyalty, service quality, student satisfaction, organizational image

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Authors: Huijing Wen, Daibao Guo

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Writing is thought to be the most complex facet of language arts. Assessing writing is difficult and subjective, and there are few scientifically validated assessments exist. Research has proposed evaluating writing using a multidimensional approach, including both qualitative and quantitative measures of handwriting, spelling and prose. Given that narrative writing has historically been a staple of literacy instruction in primary grades and is one of the three major genres Common Core State Standards required students to acquire starting in kindergarten, it is essential for teachers to understand how to measure beginning writers writing development and sources of writing difficulties through narrative writing. Guided by the theoretical models of early written expression and using empirical data, this study examines ways teachers can enact a comprehensive approach to understanding beginning writer’s narrative writing through three writing rubrics developed for a Curriculum-based Measurement (CBM). The goal is to help classroom teachers structure a framework for assessing early writing in primary classrooms. Participants in this study included 380 first-grade students from 50 classrooms in 13 schools in three school districts in a Mid-Atlantic state. Three writing tests were used to assess first graders’ writing skills in relation to both transcription (i.e., handwriting fluency and spelling tests) and translational skills (i.e., a narrative prompt). First graders were asked to respond to a narrative prompt in 20 minutes. Grounded in theoretical models of earlier expression and empirical evidence of key contributors to early writing, all written samples to the narrative prompt were coded three ways for different dimensions of writing: length, quality, and genre elements. To measure the quality of the narrative writing, a traditional holistic rating rubric was developed by the researchers based on the CCSS and the general traits of good writing. Students' genre knowledge was measured by using a separate analytic rubric for narrative writing. Findings showed that first-graders had emerging and limited transcriptional and translational skills with a nascent knowledge of genre conventions. The findings of the study provided support for the Not-So-Simple View of Writing in that fluent written expression, measured by length and other important linguistic resources measured by the overall quality and genre knowledge rubrics, are fundamental in early writing development. Our study echoed previous research findings on children's narrative development. The study has practical classroom application as it informs writing instruction and assessment. It offered practical guidelines for classroom instruction by providing teachers with a better understanding of first graders' narrative writing skills and knowledge of genre conventions. Understanding students’ narrative writing provides teachers with more insights into specific strategies students might use during writing and their understanding of good narrative writing. Additionally, it is important for teachers to differentiate writing instruction given the individual differences shown by our multiple writing measures. Overall, the study shed light on beginning writers’ narrative writing, indicating the complexity of early writing development.

Keywords: writing assessment, early writing, beginning writers, transcriptional skills, translational skills, primary grades, simple view of writing, writing rubrics, curriculum-based measurement

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Authors: John Zanetich

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Designing an educational activity that encourages inquiry and collaboration is key to engaging students in meaningful learning. Educational Information and Communications Technology (EICT) plays an important role in facilitating cooperative and collaborative learning in the classroom. The EICT also facilitates students’ learning and development of the critical thinking skills needed to solve real world problems. Projects and activities based on constructivism encourage students to embrace complexity as well as find relevance and joy in their learning. It also enhances the students’ capacity for creative and responsible real-world problem solving. Classroom activities based on constructivism offer students an opportunity to develop the higher–order-thinking skills of defining problems and identifying solutions. Participating in a classroom project is an activity for both acquiring experiential knowledge and applying new knowledge to practical situations. It also provides an opportunity for students to integrate new knowledge into a skill set using reflection. Classroom projects can be developed around a variety of learning objects including social media, knowledge management and learning communities. The construction of meaning through project-based learning is an approach that encourages interaction and problem-solving activities. Projects require active participation, collaboration and interaction to reach the agreed upon outcomes. Projects also serve to externalize the invisible cognitive and social processes taking place in the activity itself and in the student experience. This paper describes a classroom project designed to elicit interactions by helping students to unfreeze existing knowledge, to create new learning experiences, and then refreeze the new knowledge. Since constructivists believe that students construct their own meaning through active engagement and participation as well as interactions with others. knowledge management can be used to guide the exchange of both tacit and explicit knowledge in interpersonal interactions between students and guide the construction of meaning. This paper uses an action research approach to the development of a classroom project and describes the use of technology, social media and the active use of tacit knowledge in the college classroom. In this project, a closed group Facebook page becomes the virtual classroom where interaction is captured and measured using engagement analytics. In the virtual learning community, the principles of knowledge management are used to identify the process and components of the infrastructure of the learning process. The project identifies class member interests and measures student engagement in a learning community by analyzing regular posting on the Facebook page. These posts are used to foster and encourage interactions, reflect a student’s interest and serve as reaction points from which viewers of the post convert the explicit information in the post to implicit knowledge. The data was collected over an academic year and was provided, in part, by the Google analytic reports on Facebook and self-reports of posts by members. The results support the use of active tacit knowledge activities, knowledge management and social media to enhance the student learning experience and help create the knowledge that will be used by students to construct meaning.

Keywords: constructivism, knowledge management, tacit knowledge, social media

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Authors: Yi H. Wong, Li L. Chan, Chee O. Leong, Stephen Ambu, Joon W. Mak, Priyadashi S. Sahu

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Background: Acanthamoeba is a free-living opportunistic protist which is ubiquitously distributed in the environment. Virulent Acanthamoeba can cause fatal encephalitis in immunocompromised patients and potential blinding keratitis in immunocompetent contact lens wearers. Approximately 24 species have been identified but only the A. castellanii, A. polyphaga and A. culbertsoni are commonly associated with human infections. Until to date, the precise molecular basis for Acanthamoeba pathogenesis remains unclear. Previous studies reported that Acanthamoeba virulence can be diminished through prolonged axenic culture but revived through serial mouse passages. As no clear explanation on this reversible pathogenesis is established, hereby, we postulate that the epigenetic regulators, DNA-methyltransferases (DNMT) and histone-deacetylases (HDAC), could possibly be involved in granting the virulence plasticity of Acanthamoeba spp. Methods: Four rounds of mouse passages were conducted to revive the virulence potential of the virulence-attenuated Acanthamoeba castellanii strain (ATCC 50492). Briefly, each mouse (n=6/group) was inoculated intraperitoneally with Acanthamoebae cells (2x 105 trophozoites/mouse) and incubated for 2 months. Acanthamoebae cells were isolated from infected mouse organs by culture method and subjected to subsequent mouse passage. In vitro cytopathic, encystment and gelatinolytic assays were conducted to evaluate the virulence characteristics of Acanthamoebae isolates for each passage. PCR primers which targeted on the 2 members (DNMT1 and DNMT2) and 5 members (HDAC1 to 5) of the DNMT and HDAC gene families respectively were custom designed. Quantitative real-time PCR (qPCR) was performed to detect and quantify the relative expression of the two gene families in each Acanthamoeba isolates. Beta-tubulin of A. castellanii (Genbank accession no: XP_004353728) was included as housekeeping gene for data normalisation. PCR mixtures were also analyzed by electrophoresis for amplicons detection. All statistical analyses were performed using the paired one-tailed Student’s t test. Results: Our pathogenicity tests showed that the virulence-reactivated Acanthamoeba had a higher degree of cytopathic effect on vero cells, a better resistance to encystment challenge and a higher gelatinolytic activity which was catalysed by serine protease. qPCR assay showed that DNMT1 expression was significantly higher in the virulence-reactivated compared to the virulence-attenuated Acanthamoeba strain (p ≤ 0.01). The specificity of primers which targeted on DNMT1 was confirmed by sequence analysis of PCR amplicons, which showed a 97% similarity to the published DNA-methyltransferase gene of A. castellanii (GenBank accession no: XM_004332804.1). Out of the five primer pairs which targeted on the HDAC family genes, only HDAC4 expression was significantly difference between the two variant strains. In contrast to DNMT1, HDAC4 expression was much higher in the virulence-attenuated Acanthamoeba strain. Conclusion: Our mouse passages had successfully restored the virulence of the attenuated strain. Our findings suggested that DNA-methyltransferase (DNMT1) and histone deacetylase (HDAC4) expressions are associated with virulence potential of Acanthamoeba spp.

Keywords: acanthamoeba, DNA-methyltransferase, histone deacetylase, virulence-associated proteins

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Authors: Selin Guney, Andres Riquelme

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Among the main purposes of optimal and efficient forecasts of agricultural commodity prices is to guide the firms to advance the economic decision making process such as planning business operations and marketing decisions. Governments are also the beneficiaries and suppliers of agricultural price forecasts. They use this information to establish a proper agricultural policy, and hence, the forecasts affect social welfare and systematic errors in forecasts could lead to a misallocation of scarce resources. Various empirical approaches have been applied to forecast commodity prices that have used different methodologies. Most commonly-used approaches to forecast commodity sectors depend on classical time series models that assume values of the response variables are precise which is quite often not true in reality. Recently, this literature has mostly evolved to a consideration of fuzzy time series models that provide more flexibility in terms of the classical time series models assumptions such as stationarity, and large sample size requirement. Besides, fuzzy modeling approach allows decision making with estimated values under incomplete information or uncertainty. A number of fuzzy time series models have been developed and implemented over the last decades; however, most of them are not appropriate for forecasting repeated and nonconsecutive transitions in the data. The modeling scheme used in this paper eliminates this problem by introducing Markov modeling approach that takes into account both the repeated and nonconsecutive transitions. Also, the determination of length of interval is crucial in terms of the accuracy of forecasts. The problem of determining the length of interval arbitrarily is overcome and a methodology to determine the proper length of interval based on the distribution or mean of the first differences of series to improve forecast accuracy is proposed. The specific purpose of this paper is to propose and investigate the potential of a new forecasting model that integrates methodologies for determining the proper length of interval based on the distribution or mean of the first differences of series and Fuzzy Time Series- Markov Chain model. Moreover, the accuracy of the forecasting performance of proposed integrated model is compared to different univariate time series models and the superiority of proposed method over competing methods in respect of modelling and forecasting on the basis of forecast evaluation criteria is demonstrated. The application is to daily corn and soybean prices observed at three commercially important North Carolina markets; Candor, Cofield and Roaring River for corn and Fayetteville, Cofield and Greenville City for soybeans respectively. One main conclusion from this paper is that using fuzzy logic improves the forecast performance and accuracy; the effectiveness and potential benefits of the proposed model is confirmed with small selection criteria value such MAPE. The paper concludes with a discussion of the implications of integrating fuzzy logic and nonarbitrary determination of length of interval for the reliability and accuracy of price forecasts. The empirical results represent a significant contribution to our understanding of the applicability of fuzzy modeling in commodity price forecasts.

Keywords: commodity, forecast, fuzzy, Markov

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Authors: Wondale Getinet Alemu, Lillian Mwanri, Clemence Due, Telake Azale, Anna Ziersch

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Background: Mental illness is one of the most severe, chronic, and disabling public health problems that affect patients' Quality of life (QoL). Improving the QoL for people with mental illness is one of the most critical steps in stopping disease progression and avoiding complications of mental illness. Therefore, we aimed to assess the QoL and its determinants in patients with mental illness in outpatient clinics in Northwest Ethiopia in 2023. Methods: A facility-based cross-sectional study was conducted among people with mental illness in an outpatient clinic in Ethiopia. The sampling interval was decided by dividing the total number of study participants who had a follow-up appointment during the data collection period (2400) by the total sample size of 638, with the starting point selected by lottery method. The interviewer-administered WHOQOL BREF-26 tool was used to measure the QoL of people with mental illness. The domains and Health-Related Quality of Life (HRQoL) were identified. The indirect and direct effects of variables were calculated using structural equation modeling with SPSS-28 and Amos-28 software. A p-value of < 0.05 and a 95% CI were used to evaluate statistical significance. Results: A total of 636 (99.7%) participants responded and completed the WHOQOL-BREF questionnaire. The mean score of overall HRQoL of people with mental illness in the outpatient clinic was (49.6 ± 10 Sd). The highest QoL was found in the physical health domain (50.67 ±9.5 Sd), and the lowest mean QoL was found in the psychological health domain (48.41±10 Sd). Rural residents, drug nonadherence, suicidal ideation, not getting counseling, moderate or severe subjective severity, the family does not participate in patient care, and a family history of mental illness had an indirect negative effect on HRQoL. Alcohol use and psychological health domain had a direct positive effect on QoL. Furthermore, objective severity of illness, having low self-esteem, and having a history of mental illness in the family had both direct and indirect effects on QoL. Furthermore, sociodemographic factors (residence, educational status, marital status), social support-related factors (self-esteem, family not participating in patient care), substance use factors (alcohol use, tobacco use,) and clinical factors (objective and subjective severity of illness, not getting counseling, suicidal ideation, number of episodes, comorbid illness, family history of mental illness, poor drug adherence) directly and indirectly affected QoL. Conclusions: In this study, the QoL of people with mental illness was poor, with the psychological health domain being the most affected. Sociodemographic factors, social support-related factors, drug use factors, and clinical factors directly and indirectly, affect QoL through the mediator variables of physical health domains, psychological health domains, social relation health domains, and environmental health domains. In order to improve the QoL of people with mental illnesses, we recommend that emphasis be given to addressing the scourge of mental health, including the development of policy and practice drivers that address the above-identified factors.

Keywords: quality of life, mental wellbeing, mental illness, mental disorder, Ethiopia

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Authors: Carolina Cuéllar, Paz González

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Research has shown that school leadership has a significant indirect effect on students’ achievements. In Chile, evidence has also revealed that this impact is stronger in vulnerable schools. With the aim of strengthening school leadership, public policy has taken up the challenge of enhancing capabilities of novice principals through the implementation of induction programs, which include a mentoring component, entrusting the task of delivering these programs to universities. The importance of using mentoring or coaching models in the preparation of novice school leaders has been emphasized in the international literature. Thus, it can be affirmed that building leadership capacity through partnership is crucial to facilitate cognitive and affective support required in the initial phase of the principal career, gain role clarification and socialization in context, stimulate reflective leadership practice, among others. In Chile, mentoring is a recent phenomenon in the field of school leadership and it is even more new in the preparation of new principals who work in public schools. This study, funded by the Chilean Ministry of Education, sought to explore the challenges and lessons arising from the design and implementation of mentoring processes which are part of the induction programs, according to the perception of the different actors involved: ministerial agents, university coordinators, mentors and novice principals. The investigation used a qualitative design, based on a study of three cases (three induction programs). The sources of information were 46 semi-structured interviews, applied in two moments (at the beginning and end of mentoring). Content analysis technique was employed. Data focused on the uniqueness of each case and the commonalities within the cases. Five main challenges and lessons emerged in the design and implementation of mentoring within the induction programs for new principals from Chilean public schools. They comprised the need of (i) developing a shared conceptual framework on mentoring among the institutions and actors involved, which helps align the expectations for the mentoring component within the induction programs, along with assisting in establishing a theory of action of mentoring that is relevant to the public school context; (ii) recognizing trough actions and decisions at different levels that the role of a mentor differs from the role of a principal, which challenge the idea that an effective principal will always be an effective mentor; iii) improving mentors’ selection and preparation processes trough the definition of common guiding criteria to ensure that a mentor takes responsibility for developing critical judgment of novice principals, which implies not limiting the mentor’s actions to assist in the compliance of prescriptive practices and standards; (iv) generating common evaluative models with goals, instruments and indicators consistent with the characteristics of mentoring processes, which helps to assess expected results and impact; and (v) including the design of a mentoring structure as an outcome of the induction programs, which helps sustain mentoring within schools as a collective professional development practice. Results showcased interwoven elements that entail continuous negotiations at different levels. Taking action will contribute to policy efforts aimed at professionalizing the leadership role in public schools.

Keywords: induction programs, mentoring, novice principals, school leadership preparation

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Authors: C. Fernandez-Plaza, I. Abad, E. Diaz, I. Diaz

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Births occurring before the 37th week of gestation are considered preterm births. A threat of preterm is defined as the beginning of regular uterine contractions, dilation and cervical effacement between 23 and 36 gestation weeks. To author's best knowledge, the factors that determine the beginning of the birth are not completely defined yet. In particular, the incidence of sleep habits on preterm births is weekly studied. The aim of this study is to develop a model to predict the factors affecting premature delivery on pregnancy, based on the above potential risk factors, including those derived from sleep habits and light exposure at night (introduced as 12 variables obtained by a telephone survey using two questionnaires previously used by other authors). Thus, three groups of variables were included in the study (maternal, fetal and sleep habits). The study was approved by Research Ethics Committee of the Principado of Asturias (Spain). An observational, retrospective and descriptive study was performed with 481 births between January 1, 2015 and May 10, 2016 in the University Central Hospital of Asturias (Spain). A statistical analysis using SPSS was carried out to compare qualitative and quantitative variables between preterm and term delivery. Chi-square test qualitative variable and t-test for quantitative variables were applied. Statistically significant differences (p < 0.05) between preterm vs. term births were found for primiparity, multi-parity, kind of conception, place of residence or premature rupture of membranes and interruption during nights. In addition to the statistical analysis, machine learning methods to look for a prediction model were tested. In particular, tree based models were applied as the trade-off between performance and interpretability is especially suitable for this study. C5.0, recursive partitioning, random forest and tree bag models were analysed using caret R-package. Cross validation with 10-folds and parameter tuning to optimize the methods were applied. In addition, different noise reduction methods were applied to the initial data using NoiseFiltersR package. The best performance was obtained by C5.0 method with Accuracy 0.91, Sensitivity 0.93, Specificity 0.89 and Precision 0.91. Some well known preterm birth factors were identified: Cervix Dilation, maternal BMI, Premature rupture of membranes or nuchal translucency analysis in the first trimester. The model also identifies other new factors related to sleep habits such as light through window, bedtime on working days, usage of electronic devices before sleeping from Mondays to Fridays or change of sleeping habits reflected in the number of hours, in the depth of sleep or in the lighting of the room. IF dilation < = 2.95 AND usage of electronic devices before sleeping from Mondays to Friday = YES and change of sleeping habits = YES, then preterm is one of the predicting rules obtained by C5.0. In this work a model for predicting preterm births is developed. It is based on machine learning together with noise reduction techniques. The method maximizing the performance is the one selected. This model shows the influence of variables related to sleep habits in preterm prediction.

Keywords: machine learning, noise reduction, preterm birth, sleep habit

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Authors: Dana Zelicha

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A 6-week mindfulness programme was launched to improve the well being and performance of 20 managers (including the supervisor) of an international corporation in London. A unique assessment methodology was customised to the organisation’s needs, measuring four parameters: prioritising skills, listening skills, mindfulness levels and happiness levels. All parameters showed significant improvements (p < 0.01) post intervention, with a remarkable increase in listening skills and mindfulness levels. Although corporate mindfulness programmes have proven to be effective, the challenge remains the low engagement levels at home and the implementation of these tools beyond the scope of the intervention. This study has offered an innovative approach to enforce home engagement levels, which yielded promising results. The programme launched with a 2-day introduction intervention, which was followed by a 6-week training course (1 day a week; 2 hours each). Participants learned all basic principles of mindfulness such as mindfulness meditations, Mindfulness Based Stress Reduction (MBSR) techniques and Mindfulness Based Cognitive Therapy (MBCT) practices to incorporate into their professional and personal lives. The programme contained experiential mindfulness meditations and innovative mindfulness tools (OWBA-MT) created by OWBA - The Well Being Agency. Exercises included Mindful Meetings, Unitasking and Mindful Feedback. All sessions concluded with guided discussions and group reflections. One fundamental element of this programme was engagement level outside of the workshop. In the office, participants connected with a mindfulness buddy - a team member in the group with whom they could find support throughout the programme. At home, participants completed online daily mindfulness forms that varied according to weekly themes. These customised forms gave participants the opportunity to reflect on whether they made time for daily mindfulness practice, and to facilitate a sense of continuity and responsibility. At the end of the programme, the most engaged team member was crowned the ‘mindful maven’ and received a special gift. The four parameters were measured using online self-reported questionnaires, including the Listening Skills Inventory (LSI), Mindfulness Attention Awareness Scale (MAAS), Time Management Behaviour Scale (TMBS) and a modified version of the Oxford Happiness Questionnaire (OHQ). Pre-intervention questionnaires were collected at the start of the programme, and post-intervention data was collected 4-weeks following completion. Quantitative analysis using paired T-tests of means showed significant improvements, with a 23% increase in listening skills, a 22% improvement in mindfulness levels, a 12% increase in prioritising skills, and an 11% improvement in happiness levels. Participant testimonials exhibited high levels of satisfaction and the overall results indicate that the mindfulness programme substantially impacted the team. These results suggest that 6-week mindfulness programmes can improve employees’ capacities to listen and work well with others, to effectively manage time and to experience enhanced satisfaction both at work and in life. Limitations noteworthy to consider include the afterglow effect and lack of generalisability, as this study was conducted on a small and fairly homogenous sample.

Keywords: corporate mindfulness, listening skills, organisational well being, prioritising skills, mindful leadership

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Authors: Ved Vrat Verma, Rani Gupta, Manisha Goel

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γ-glutamyltranspeptidase (GGT: EC 2.3.2.2) is an N-terminal nucleophile hydrolase conserved in all three domains of life. GGT plays a key role in glutathione metabolism where it catalyzes the breakage of the γ-glutamyl bonds and transfer of γ-glutamyl group to water (hydrolytic activity) or amino acids or short peptides (transpeptidase activity). GGTs from bacteria, archaea, and eukaryotes (human, rat and mouse) are homologous proteins sharing >50% sequence similarity and conserved four layered αββα sandwich like three dimensional structural fold. These proteins though similar in their structure to each other, are quite diverse in their enzyme activity: some GGTs are better at hydrolysis reactions but poor in transpeptidase activity, whereas many others may show opposite behaviour. GGT is known to be involved in various diseases like asthma, parkinson, arthritis, and gastric cancer. Its inhibition prior to chemotherapy treatments has been shown to sensitize tumours to the treatment. Microbial GGT is known to be a virulence factor too, important for the colonization of bacteria in host. However, all known inhibitors (mimics of its native substrate, glutamate) are highly toxic because they interfere with other enzyme pathways. However, a few successful efforts have been reported previously in designing species specific inhibitors. We aim to leverage the diversity seen in GGT family (pathogen vs. eukaryotes) for designing specific inhibitors. Thus, in the present study, we have used DIVERGE software to identify sites in GGT proteins, which are crucial for the functional and structural divergence of these proteins. Since, type II divergence sites vary in clade specific manner, so type II divergent sites were our focus of interest throughout the study. Type II divergent sites were identified for pathogen vs. eukaryotes clusters and sites were marked on clade specific representative structures HpGGT (2QM6) and HmGGT (4ZCG) of pathogen and eukaryotes clade respectively. The crucial divergent sites within 15 A radii of the binding cavity were highlighted, and in-silico mutations were performed on these sites to delineate the role of these sites on the mechanism of catalysis and protein folding. Further, the amino acid network (AAN) analysis was also performed by Cytoscape to delineate assortative mixing for cavity divergent sites which could strengthen our hypothesis. Additionally, molecular dynamics simulations were performed for wild complexes and mutant complexes close to physiological conditions (pH 7.0, 0.1 M ionic strength and 1 atm pressure) and the role of putative divergence sites and structural integrities of the homologous proteins have been analysed. The dynamics data were scrutinized in terms of RMSD, RMSF, non-native H-bonds and salt bridges. The RMSD, RMSF fluctuations of proteins complexes are compared, and the changes at protein ligand binding sites were highlighted. The outcomes of our study highlighted some crucial divergent sites which could be used for novel inhibitors designing in a species-specific manner. Since, for drug development, it is challenging to design novel drug by targeting similar protein which exists in eukaryotes, so this study could set up an initial platform to overcome this challenge and help to deduce the more effective targets for novel drug discovery.

Keywords: γ-glutamyltranspeptidase, divergence, species-specific, drug design

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Authors: Adrienn Kiss, Karoly Zauer, Gyorgy Keglevich, Rita Molnarne Bernath

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Ginger (Zingiber officinale) is a perennial plant from Southeast Asia, widely used as a spice, herb, and medicine for many illnesses since its beneficial health effects were observed thousands of years ago. Among the compounds found in ginger, zingerone [4-hydroxy-3- methoxyphenyl-2-butanone] deserves special attention: it has an anti-inflammatory and antispasmodic effect, it can be used in case of diarrheal disease, helps to prevent the formation of blood clots, has antimicrobial properties, and can also play a role in preventing the Alzheimer's disease. Ferulic acid [(E)-3-(4-hydroxy-3-methoxyphenyl)-prop-2-enoic acid] is another cinnamic acid derivative in ginger, which has promising properties. Like many phenolic compounds, ferulic acid is also an antioxidant. Based on the results of animal experiments, it is assumed to have a direct antitumoral effect in lung and liver cancer. It also deactivates free radicals that can damage the cell membrane and the DNA and helps to protect the skin against UV radiation. The aim of this work was to synthesize these two compounds by new methods. A few of the reactions were based on the hydrogenation of dehydrozingerone [4-(4-Hydroxy-3-methoxyphenyl)-3-buten-2-one] to zingerone. Dehydrozingerone can be synthesized by a relatively simple method from acetone and vanillin with good yield (80%, melting point: 41 °C). Hydrogenation can be carried out chemically, for example by the reaction of zinc and acetic acid, or Grignard magnesium and ethyl alcohol. Another way to complete the reduction is the electrochemical pathway. The electrolysis of dehydrozingerone without diaphragm in aqueous media was attempted to produce ferulic acid in the presence of sodium carbonate and potassium iodide using platinum electrodes. The electrolysis of dehydrozingerone in the presence of potassium carbonate and acetic acid to prepare zingerone was carried out similarly. Ferulic acid was expected to be converted to dihydroferulic acid [3-(4-Hydroxy-3-methoxyphenyl)propanoic acid] in potassium hydroxide solution using iron electrodes, separating the anode and cathode space with a Soxhlet paper sheath impregnated with saturated magnesium chloride solution. For this reaction, ferulic acid was synthesized from vanillin and malonic acid in the presence of pyridine and piperidine (yield: 88.7%, melting point: 173°C). Unfortunately, in many cases, the expected transformations did not happen or took place in low conversions, although gas evolution occurred. Thus, a deeper understanding of these experiments and optimization are needed. Since both compounds are found in different plants, they can also be obtained by alkaline extraction or steam distillation from distinct plant parts (ferulic acid from ground bamboo shoots, zingerone from grated ginger root). The products of these reactions are rich in several other organic compounds as well; therefore, their separation must be solved to get the desired pure material. The products of the reactions described above were characterized by infrared spectral data and melting points. The use of these two simple methods may be informative for the formation of the products. In the future, we would like to study the ferulic acid and zingerone content of other plants and extract them efficiently. The optimization of electrochemical reactions and the use of other test methods are also among our plans.

Keywords: ferulic acid, ginger, synthesis, zingerone

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Authors: Paola Montenegro, Maria de los Angeles Balaguera Villa

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In February 2022 Colombia achieved a historic milestone in ensuring universal access to abortion rights with ruling C-055 of 2022 decriminalising abortion up to 24 weeks of gestation. In the context of this triumph and the expansion of telemedicine services in the wake of the COVID-19 pandemic, this research studied the acceptability of self-care abortion in young people (13 - 28 years) through a telemedicine service and also explored the primary needs that should be the focus of such care. The results shine light on a more comprehensive understanding of opportunities and challenges of teleabortion practices in a context that combines overall higher access to technology and low access to reliable information of safe abortion, stigma, and scarcity especially felt by transnational migrants, racialised people, trans men and non-binary people. Through a mixed methods approach, this study collected 5.736 responses to a virtual survey disseminated nationwide in Colombia and 47 in-person interviews (24 of them with people who were assigned female at birth and 21 with local key stakeholders in the abortion ecosystem). Quantitative data was analyzed using Stata SE Version 16.0 and qualitative analysis was completed through NVivo using thematic analysis. Key findings of the research suggest that self-care abortion is practice with growing acceptability among young people, but important adjustments must be made to meet quality of care expectations of users. Elements like quick responses from providers, lower costs, and accessible information were defined by users as decisive factors to choose over the abortion service provider. In general, the narratives in participants about quality care were centred on the promotion of autonomy and the provision of accompaniment and care practices, also perceived as transformative and currently absent of most health care services. The most staggering findings from the investigation are related to current barriers faced by young people in abortion contexts even when the legal barriers have: high rates of scepticism and distrust associated with pitfalls of telehealth and structural challenges associated with lacking communications infrastructure, among a few of them. Other important barriers to safe self-care abortion identified by participants surfaced like lack of privacy and confidentiality (especially in rural areas of the country), difficulties accessing reliable information, high costs of procedures and expenses related to travel costs or having to cease economic activities, waiting times, and stigma are among the primary barriers to abortion identified by participants. Especially in a scenario marked by unprecedented social, political and economic disruptions due to the COVID-19 pandemic, the commitment to design better care services that can be adapted to the identities, experiences, social contexts and possibilities of the user population is more necessary than ever. In this sense, the possibility of expanding access to services through telemedicine brings us closer to the opportunity to rethink the role of health care models in transforming the role of individuals and communities to make autonomous, safe and informed decisions about their own health and well-being.

Keywords: contraception, family planning, premarital fertility, unplanned pregnancy

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Authors: Aleksandra Bylinska, Samantha Slight-Webb, Kevin Thomas, Miles Smith, Susan Macwana, Nicolas Dominguez, Eliza Chakravarty, Joan T. Merrill, Judith A. James, Joel M. Guthridge

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Background: Systemic Lupus Erythematosus (SLE) is an interferon-related autoimmune disease characterized by B cell dysfunction. One of the main hallmarks is a loss of tolerance to self-antigens leading to increased levels of autoantibodies against nuclear components (ANAs). However, up to 20% of healthy ANA+ individuals will not develop clinical illness. SLE is more prevalent among women and minority populations (African, Asian American and Hispanics). Moreover, African Americans have a stronger interferon (IFN) signature and develop more severe symptoms. The exact mechanisms involved in ethnicity-dependent B cell dysregulation and the progression of autoimmune disease from ANA+ healthy individuals to clinical disease remains unclear. Methods: Peripheral blood mononuclear cells (PBMCs) from African (AA) and European American (EA) ANA- (n=12), ANA+ (n=12) and SLE (n=12) individuals were assessed by multimodal scRNA-Seq/CITE-Seq methods to examine differential gene signatures in specific B cell subsets. Library preparation was done with a 10X Genomics Chromium according to established protocols and sequenced on Illumina NextSeq. The data were further analyzed for distinct cluster identification and differential gene signatures in the Seurat package in R and pathways analysis was performed using Ingenuity Pathways Analysis (IPA). Results: Comparing all subjects, 14 distinct B cell clusters were identified using a community detection algorithm and visualized with Uniform Manifold Approximation Projection (UMAP). The proportion of each of those clusters varied by disease status and ethnicity. Transitional B cells trended higher in ANA+ healthy individuals, especially in AA. Ribonucleoprotein high population (HNRNPH1 elevated, heterogeneous nuclear ribonucleoprotein, RNP-Hi) of proliferating Naïve B cells were more prevalent in SLE patients, specifically in EA. Interferon-induced protein high population (IFIT-Hi) of Naive B cells are increased in EA ANA- individuals. The proportion of memory B cells and plasma cells clusters tend to be expanded in SLE patients. As anticipated, we observed a higher signature of cytokine-related pathways, especially interferon, in SLE individuals. Pathway analysis among AA individuals revealed an NRF2-mediated Oxidative Stress response signature in the transitional B cell cluster, not seen in EA individuals. TNFR1/2 and Sirtuin Signaling pathway genes were higher in AA IFIT-Hi Naive B cells, whereas they were not detected in EA individuals. Interferon signaling was observed in B cells in both ethnicities. Oxidative phosphorylation was found in age-related B cells (ABCs) for both ethnicities, whereas Death Receptor Signaling was found only in EA patients in these cells. Interferon-related transcription factors were elevated in ABCs and IFIT-Hi Naive B cells in SLE subjects of both ethnicities. Conclusions: ANA+ healthy individuals have altered gene expression pathways in B cells that might drive apoptosis and subsequent clinical autoimmune pathogenesis. Increases in certain regulatory pathways may delay progression to SLE. Further, AA individuals have more elevated activation pathways that may make them more susceptible to SLE.

Keywords:

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Authors: Teresa Wanjiru Mbatia

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On 7th June 2011, the government Minister of Roads in Kenya announced the proposed construction of a major highway known as a southern bypass to run on the northern border of the Nairobi National Park. The following day on 8th June 2011, the chairperson of the Friends of Nairobi National Park (FONNAP) posted a protest statement on their website, with the heading, ‘Nairobi Park is Not a cake’ alerting its members and conservation groups, with the aim of getting support to the campaign against the government’s intention to hive off a section of the park for road construction. This was the first and earliest statement that led to a series of other events that culminated in conservationists and some other members of the public campaign against the government’s plan to hive off sections of the park to build road and railway infrastructure in or around the park. Together with other non-state actors, mostly non-governmental organisations in conservation/environment and tourism businesses, FoNNAP issued a series of other statements on social, print and electronic media to battle against road and railway construction. This paper examined the strategies, outcomes and interests of actors involved in opposing/proposing the development of transport infrastructure in and around the Nairobi National Park. Specifically, the objectives were to analyse the: (1) Arguments put forward by the eco-warriors to protest infrastructure development; (2) Background and interests of the eco-warriors; (3) Needs/interests and opinions of ordinary common citizens on transport infrastructural development, particularly in and around the urban nature reserve and (4) Final outcomes of the eco-politics surrounding infrastructure development in and around Nairobi National Park. The methodological approach used was environmental history and the social construction of nature. The study collected combined qualitative data using four main approaches, the grounded theory approach, narratives, case studies and a phenomenological approach. The information collected was analysed using critical discourse analysis. The major findings of the study were that under the guise of “public participation,” influential non-state actors have the capacity to perpetuate social-spatial inequalities in the form of curtailing the majority from accessing common public goods. A case in point in this study is how the efforts of powerful conservationists, environmentalists, and tourism businesspersons managed to stall the construction of much-needed road and railway infrastructure severally through litigations in lengthy environmental court processes involving injunctions and stop orders to the government bodies in charge. Moreover, powerful non-state actors were found to have formed informal and sometimes formal coalitions with politicians with selfish interests, which serves to deepen the exclusionary practices and the common good. The study concludes that mostly composed of certain types of elites (NGOs, business communities, politicians and privileged social-cultural groups), non-state actors have used participatory policies to advance their own interests at the expense of the majority whom they claim to represent. These practices are traced to the historically unjust social, political, and economic forces involved in the production of space in Nairobi.

Keywords: eco-politics, exclusion, infrastructure, Nairobi national park, non-state actors, protests

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Authors: P. J. Sweeney, T. Ahtoniemi, J. Puoliväli, T. Laitinen, K. Lehtimäki, A. Nurmi, D. Wells

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Duchenne muscular dystrophy (DMD) is caused by an X linked mutation in the dystrophin gene; lack of dystrophin causes a progressive muscle necrosis which leads to a progressive decrease in mobility in those suffering from the disease. The MDX mouse, a mutant mouse model which displays a frank dystrophinopathy, is currently widely employed in pre clinical efficacy models for treatments and therapies aimed at DMD. In general the end-points examined within this model have been based on invasive histopathology of muscles and serum biochemical measures like measurement of serum creatine kinase (sCK). It is established that a “critical period” between 4 and 6 weeks exists in the MDX mouse when there is extensive muscle damage that is largely sub clinical but evident with sCK measurements and histopathological staining. However, a full characterization of the MDX model remains largely incomplete especially with respect to the ability to aggravate of the muscle damage beyond the critical period. The purpose of this study was to attempt to aggravate the muscle damage in the MDX mouse and to create a wider, more readily translatable and discernible, therapeutic window for the testing of potential therapies for DMD. The study consisted of subjecting 15 male mutant MDX mice and 15 male wild-type mice to an intense chronic exercise regime that consisted of bi-weekly (two times per week) treadmill sessions over a 12 month period. Each session was 30 minutes in duration and the treadmill speed was gradually built up to 14m/min for the entire session. Baseline plasma creatine kinase (pCK), treadmill training performance and locomotor activity were measured after the “critical period” at around 10 weeks of age and again at 14 weeks of age, 6 months, 9 months and 12 months of age. In addition, kinematic gait analysis was employed using a novel analysis algorithm in order to compare changes in gait and fine motor skills in diseased exercised MDX mice compared to exercised wild type mice and non exercised MDX mice. In addition, a morphological and metabolic profile (including lipid profile), from the muscles most severely affected, the gastrocnemius muscle and the tibialis anterior muscle, was also measured at the same time intervals. Results indicate that by aggravating or exacerbating the underlying muscle damage in the MDX mouse by exercise a more pronounced and severe phenotype in comes to light and this can be picked up earlier by kinematic gait analysis. A reduction in mobility as measured by open field is not apparent at younger ages nor during the critical period, but changes in gait are apparent in the mutant MDX mice. These gait changes coincide with pronounced morphological and metabolic changes by non-invasive anatomical MRI and proton spectroscopy (1H-MRS) we have reported elsewhere. Evidence of a progressive asymmetric pathology in imaging parameters as well as in the kinematic gait analysis was found. Taken together, the data show that chronic exercise regime exacerbates the muscle damage beyond the critical period and the ability to measure through non-invasive means are important factors to consider when performing preclinical efficacy studies in the MDX mouse.

Keywords: Gait, muscular dystrophy, Kinematic analysis, neuromuscular disease

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Authors: M. Naveed Yasin, Robert Brooke, Andrew Chan, Geoffrey I. N. Waterhouse, Drew Evans, Darren Svirskis, Ilva D. Rupenthal

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Poly(3,4-ethylenedioxythiophene) (PEDOT) is a robust conducting polymer (CP) exhibiting high conductivity and environmental stability. It can be synthesized by either chemical, electrochemical or vapour phase polymerization (VPP). Dexamethasone sodium phosphate (dexP) is an anionic drug molecule which has previously been loaded onto PEDOT as a dopant via electrochemical polymerisation; however this technique requires conductive surfaces from which polymerization is initiated. On the other hand, VPP produces highly organized biocompatible CP structures while polymerization can be achieved onto a range of surfaces with a relatively straight forward scale-up process. Following VPP of PEDOT, dexP can be loaded and subsequently released via ion-exchange. This study aimed at preparing and characterising both non-porous and porous VPP PEDOT structures including examining drug loading and release via ion-exchange. Porous PEDOT structures were prepared by first depositing a sacrificial polystyrene (PS) colloidal template on a substrate, heat curing this deposition and then spin coating it with the oxidant solution (iron tosylate) at 1500 rpm for 20 sec. VPP of both porous and non-porous PEDOT was achieved by exposing to monomer vapours in a vacuum oven at 40 mbar and 40 °C for 3 hrs. Non-porous structures were prepared similarly on the same substrate but without any sacrificial template. Surface morphology, compositions and behaviour were then characterized by atomic force microscopy (AFM), scanning electron microscopy (SEM), x-ray photoelectron spectroscopy (XPS) and cyclic voltammetry (CV) respectively. Drug loading was achieved by 50 CV cycles in a 0.1 M dexP aqueous solution. For drug release, each sample was exposed to 20 mL of phosphate buffer saline (PBS) placed in a water bath operating at 37 °C and 100 rpm. Film was stimulated (continuous pulse of ± 1 V at 0.5 Hz for 17 mins) while immersed into PBS. Samples were collected at 1, 2, 6, 23, 24, 26 and 27 hrs and were analysed for dexP by high performance liquid chromatography (HPLC Agilent 1200 series). AFM and SEM revealed the honey comb nature of prepared porous structures. XPS data showed the elemental composition of the dexP loaded film surface, which related well with that of PEDOT and also showed that one dexP molecule was present per almost three EDOT monomer units. The reproducible electroactive nature was shown by several cycles of reduction and oxidation via CV. Drug release revealed success in drug loading via ion-exchange, with stimulated porous and non-porous structures exhibiting a proof of concept burst release upon application of an electrical stimulus. A similar drug release pattern was observed for porous and non-porous structures without any significant statistical difference, possibly due to the thin nature of these structures. To our knowledge, this is the first report to explore the potential of VPP prepared PEDOT for stimuli-responsive drug delivery via ion-exchange. The produced porous structures were ordered and highly porous as indicated by AFM and SEM. These porous structures exhibited good electroactivity as shown by CV. Future work will investigate porous structures as nano-reservoirs to increase drug loading while sealing these structures to minimize spontaneous drug leakage.

Keywords: PEDOT for ion-exchange drug delivery, stimuli-responsive drug delivery, template based porous PEDOT structures, vapour phase polymerization of PEDOT

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Authors: Rijnbout-St.James Willem, Lindner Volkhard, Scholand Mary Beth, Ashton M. Tillett, Di Gennaro Michael Jude, Smith Silvia Enrica

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Introduction: Fibrosing lung diseases are characterized by changes in the lung interstitium and are classified based on etiology: 1) environmental/exposure-related, 2) autoimmune-related, 3) sarcoidosis, 4) interstitial pneumonia, and 4) idiopathic. Among interstitial lung diseases (ILD) idiopathic forms, idiopathic pulmonary fibrosis (IPF) is the most severe. Pathogenesis of IPF is characterized by an increased presence of proinflammatory mediators, resulting in alveolar injury, where injury to alveolar epithelium precipitates an increase in collagen deposition, subsequently thickening the alveolar septum and decreasing gas exchange. Identifying biomarkers implicated in the pathogenesis of lung fibrosis is key to developing new therapies and improving the efficacy of existing therapies. The transforming growth factor-beta (TGF-B1), a mediator of tissue repair associated with WNT5A signaling, is partially responsible for fibroblast proliferation in ILD and is the target of Pirfenidone, one of the antifibrotic therapies used for patients with IPF. Canonical TGF-B signaling is mediated by the proteins SMAD 2/3, which are, in turn, indirectly regulated by Collagen Triple Helix Repeat Containing 1 (CTHRC1). In this study, we tested the following hypotheses: 1) CTHRC1 is more elevated in the ILD cohort compared to unaffected controls, and 2) CTHRC1 is differently expressed among ILD types. Material and Methods: CTHRC1 levels were measured by ELISA in 171 plasma samples from the deidentified University of Utah ILD cohort. Data represent a cohort of 131 ILD-affected participants and 40 unaffected controls. CTHRC1 samples were categorized by a pulmonologist based on affectation status and disease subtypes: IPF (n = 45), sarcoidosis (4), nonspecific interstitial pneumonia (16), hypersensitivity pneumonitis (n = 7), interstitial pneumonia (n=13), autoimmune (n = 15), other ILD - a category that includes undifferentiated ILD diagnoses (n = 31), and unaffected controls (n = 40). We conducted a single-factor ANOVA of plasma CTHRC1 levels to test whether CTHRC1 variance among affected and non-affected participants is statistically significantly different. In-silico analysis was performed with Ingenuity Pathway Analysis® to characterize the role of CTHRC1 in the pathway of lung fibrosis. Results: Statistical analyses of CTHRC1 in plasma samples indicate that the average CTHRC1 level is significantly higher in ILD-affected participants than controls, with the autoimmune ILD being higher than other ILD types, thus supporting our hypotheses. In-silico analyses show that CTHRC1 indirectly activates and phosphorylates SMAD3, which in turn cross-regulates TGF-B1. CTHRC1 also may regulate the expression and transcription of TGFB-1 via WNT5A and its regulatory relationship with CTNNB1. Conclusion: In-silico pathway analyses demonstrate that CTHRC1 may be an important biomarker in ILD. Analysis of plasma samples indicates that CTHRC1 expression is positively associated with ILD affectation, with autoimmune ILD having the highest average CTHRC1 values. While characterizing CTHRC1 levels in plasma can help to differentiate among ILD types and predict response to Pirfenidone, the extent to which plasma CTHRC1 level is a function of ILD severity or chronicity is unknown.

Keywords: interstitial lung disease, CTHRC1, idiopathic pulmonary fibrosis, pathway analyses

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Authors: Negar Vkilbashi, Hassan Kabiri

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Modern diplomacy, in its general form, refers to the people and not the governments, and diplomacy tactics are more addressed to the people than to the governments. Media diplomacy and cyber diplomacy are also one of the sub-branches of public diplomacy and, in fact, the role of media in the process of influencing public opinion and directing foreign policy. Mass media, including written, radio and television, theater, satellite, internet, and news agencies, transmit information and demands. What the Qatari government tried to implement in the countries of the region during the Arab Spring and after was through its important media, Al Jazeera. The embargo on Qatar began in 2017, when Saudi Arabia, the United Arab Emirates, Bahrain, and Egypt imposed a land, sea, and air blockade against the country. The media tool constitutes the cornerstone of soft power in the field of foreign policy, which Qatari leaders have consistently resorted to over the past two decades. Undoubtedly, the role it played in covering the events of the Arab Spring has created geopolitical tensions. The United Arab Emirates and other neighboring countries sometimes criticize Al Jazeera for providing a platform for the Muslim Brotherhood, Hamas, and other Islamists to promote their ideology. In 2011, at the same time as the Arab Spring, Al Jazeera reached the peak of its popularity. Al Jazeera's live coverage of protests in Tunisia, Egypt, Yemen, Libya, and Syria helped create a unified narrative of the Arab Spring, with audiences tuning in every Friday to watch simultaneous protests across the Middle East. Al Jazeera operates in three groups: First, it is a powerful base in the hands of the government so that it can direct and influence Arab public opinion. Therefore, this network has been able to benefit from the unlimited financial support of the Qatar government to promote its desired policies and culture. Second, it has provided an attractive platform for politicians and scientific and intellectual elites, thus attracting their support and defense from the government and its rulers. Third, during the last years of Prince Hamad's reign, the Al Jazeera network formed a deterrent weapon to counter the media and political struggle campaigns. The importance of the research is that this network covers a wide range of people in the Middle East and, therefore, has a high influence on the decision-making of countries. On the other hand, Al Jazeera is influential as a tool of public diplomacy and soft power in Qatar's foreign policy, and by studying it, the results of its effectiveness in the past years can be examined. Using a qualitative method, this research analyzes the impact of the media on the implementation of Qatar's foreign policy on the public opinion of the people of the Middle East. Data collection has been done by the secondary method, that is, reading related books, magazine articles, newspaper reports and articles, and analytical reports of think tanks. The most important findings of the research are that Al Jazeera plays an important role in Qatar's foreign policy in Qatar's public diplomacy. So that, in 2011, 2017 and 2023, it played an important role in Qatar's foreign policy in various crises. Also, the people of Arab countries use Al-Jazeera as their first reference.

Keywords: Al Jazeera, Qatar, media, diplomacy

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Authors: Ayoti Banerjee, Meenakshi Banerjee

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The study of metamorphic reaction textures is important in contributing to our understanding of the evolution of metamorphic terranes. Where preserved, they provide information on changes in the P-T conditions during the metamorphic history of the rock, and thus allow us to speculate on the P-T-t evolution of the terrane. Mafic dykes have attracted the attention of petrologists because they act as window to mantle. This rock represents a mafic dyke of doleritic composition. It is fine to medium grained in which clinopyroxene are enclosed by the lath shaped plagioclase grains to form spectacular ophitic texture. At places, sub ophitic texture was also observed. Grains of pyroxene and plagioclase show very less deformation typically plagioclase showing deformed lamella along with plagioclase-clinopyroxene-phyric granoblastic fabric within a groundmass of feldspar microphenocrysts and Fe–Ti oxides. Both normal and reverse zoning were noted in the plagioclase laths. The clinopyroxene grains contain exsolved phases such as orthopyroxene, plagioclase, magnetite, ilmenite along the cleavage traces and the orthopyroxene lamella form granules in the periphery of the clinopyroxene grains. Garnet corona also develops preferentially around plagioclase at the contact of clinopyroxene, ilmenite or magnetite. Tiny quartz and K-fs grains showed symplectic intergrowth with garnet at a few places. The product quartz formed along with garnet rims the coronal garnet and the reacting clinopyroxene. Thin amphibole corona formed along the periphery of deformed plagioclase and clinopyroxene occur as patches over the magmatic minerals. The amphibole coronas cannot be assigned to a late magmatic stage and are interpreted as reactive being restricted to the contact between clinopyroxene and plagioclase, thus postdating the crystallization of both. The amphibole and garnet do not share grain boundary in the entire rock and is thus pointing towards simultaneous crystallization. Olivine is absent. Spectacular myrmekitic growth of orthoclase and quartz rimming the plagioclase is consistent with the potash metasomatic effects that is also found in other rocks of this region. These textural features are consistent with a phase of fluid induced metamorphism (retrogression). But the appearance of coronal garnet and amphibole exclusive of each other reflects the participation if Ti as the prime reason. Presence of Ti as a reactant phase is a must for amphibole forming reactions whereas it is not so in case of garnet forming reactions although the reactants are the same plagioclase and clinopyroxene in both cases. These findings are well validated by petrographical and textural analysis. In order to obtain balanced chemical reactions that explain formation of amphibole and garnet in the mafic dyke rocks a matrix operation technique called Singular Value Decomposition (SVD) was adopted utilizing the measured chemical compositions of the minerals. The computer program C-Space was used for this purpose and the required compositional matrix. Data fed to C-Space was after doing cation-calculation of the oxide percentages obtained from EPMA analysis. The Garnet-Clinopyroxene geothermometer yielded a temperature of 650 degrees Celsius. The Garnet-Clinopyroxene-Plagioclase geobarometer and Al-in amphibole yielded roughly 7.5 kbar pressure.

Keywords: corona, dolerite, geothermometer, metasomatism, metamorphic reaction texture, retrogression

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Authors: Pedro Felgueiras, Ana Miguel, Nélson Almeida, Raquel Silva

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Objective: Through the study of a clinical case of delusional somatic disorder secondary to phantogeusia, we aim to highlight the importance of considering psychosomatic conditions in differential diagnosis, as well as to emphasize the complexity of its comprehension, treatment, and respective impact on patients’ functioning. Methods: Bearing this in mind, we conducted a critical analysis of a case series based on patient observations, clinical data, and complementary diagnostic methods, as well as a non-systematic review of the literature on the subject. Results: A 61-year-old female patient with no history of psychiatric conditions. Family psychiatric history of mood disorder (depression), with psychotic features found in her mother. Medical history of many comorbidities affecting different organ systems (endocrine, gastrointestinal, genitourinary, ophthalmological). Documented neuroticism traits of personality. The patient’s family described a persistent concern about several physical symptoms across her life, with a continuous effort to obtain explanations about any sensation out of her normal perception. Since being subjected to endoscopy in 2018, she started complaints of persistent phantogeusia (acid taste) and developed excessive thoughts, feelings, and behaviors associated with this somatic symptom. The patient was evaluated by several medical specialties, and an extensive panel of medical exams was carried out, excluding any disease. Besides all the investigation and with no evidence of disease signs, acute anxiety, time, and energy dispended to this symptom culminated in severe psychosocial impairment. The patient was admitted to a psychiatric ward for investigation and treatment of this clinical picture, leading to the diagnosis of the delusional somatic disorder. In order to exclude the acute organic etiology of this psychotic disorder, an analytic panel was carried out with no abnormal results. In the context of a psychotic clinical picture, a CT scan was performed, which revealed a right cortical vascular lesion. Neuropsychological evaluation was made, with the description of cognitive functioning being globally normative. During treatment with an antipsychotic (pimozide), a complete remission of the somatic delusion was associated with the disappearance of gustative perception disturbance. In follow-up, a relapse of gustative sensation was documented, and her thoughts and speech were dominated by concerns about multiple somatic symptoms. Conclusion: In terms of abnormal bodily sensations, the oral cavity is one of the frequent sites of delusional disorder. Patients with these gustatory perception distortions complain about unusual sensations without corresponding abnormal findings in the oral area. Its pathophysiology has not been fully elucidated yet. In terms of its comprehensive psychopathology, this case was hypothesized as a paranoid development of a delusional somatic disorder triggered by a post-invasive procedure phantogeusia (which is described as a possible side effect of an endoscopy) in a patient with an anankastic personality. This case presents interesting psychopathology, reinforcing the complexity of psychosomatic disorders in terms of their etiopathogenesis, clinical treatment, and long-term prognosis.

Keywords: psychosomatics, delusional somatic disorder, phantogeusia, paranoid development

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Authors: Humaira Maheen, Elizabeth Hoban, Catherine Bennette

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Background: Globally, role of Community Health Workers (CHW) as front line disaster health work force is underutilized. Developing countries which are at risk of natural disasters or humanitarian emergencies should lay down effective strategies especially to ensure adequate access to maternity care during crisis situation by using CHW as they are local, trained, and most of them possess a good relationship with the community. The Minimum Initial Service Package (MISP) is a set of universal guidelines that addresses women’s reproductive health needs during the first phase of an emergency. According to the MISP, pregnant women should have access to a skilled birth attendant and adequate transportation arrangements so they can access a maternity care facility. Pakistan is one of the few countries which has been severely affected by a number of natural disaster as well as humanitarian emergencies in last decade. Pakistan has a young and structured National Disaster Management System in place, where District Authorities play a vital role in disaster management. The District Health Department develops the contingency health plan for an emergency situation and implements it under the existing district health human resources (health workers and medical staff at the health facility) and infrastructure (health care facilities). Methods: A mixed methods study was conducted in rural villages of Sindh adjacent to the river Indus, and included in-depth interviews with 15 women who gave birth during the floods, structured interviews with 668 women who were pregnant during 2010-2014, and in-depth interviews with 25 community health workers (CHW) and 30 key informants. Results: Women said that giving birth in the relief camps during the floods was one of the most challenging times of their life. The district health department didn’t make transportation arrangement for labouring women from relief camp to the nearest health care facility. As a result 91.2% women gave birth in temporary shelters with the help of a traditional birth attendant (Dai) with no clean physical space available to birth. Of the 332 women who were pregnant at the time of the floods, 26 had adverse birth outcomes; 10 had miscarriages, 14 had stillbirths and there were four neonatal deaths. Conclusion: The district health department was not able to provide access to adequate maternity care during according to the international standard during the floods in 2011. We propose a model where CHWs will be used as frontline maternity care providers during any emergency or disaster situations in Pakistan. A separate "birthing station" should be mandatory in all district relief camps, managed by CHWs. Community midwives (CMW) would and the Lady Health Workers (LHW) would provide antenatal and postnatal care alongside, vaccination for pregnant women, neonates and children under five. There must be an ambulance facility for emergency obstetric cases and all district health facilities should have at least two medical staff identified and trained for emergency obstetric management. The District Health Department must provide clean birthing kits and regular and emergency contraceptives in the relief camps. Methods: A mixed methods study was conducted in rural villages of Sindh adjacent to the river Indus, and included in-depth interviews with 15 women who gave birth during the floods, structured interviews with 668 women who were pregnant during 2010-2014, and in-depth interviews with 25 community health workers (CHW) and 30 key informants. Results: Women said that giving birth in the relief camps during the floods was one of the most challenging times of their life. Nearly 91.2% women gave birth in temporary shelters with the help of a traditional birth attendant (Dai) with no clean physical space available to birth, and the health camp was mostly accessed by men and always overcrowded. There was no obstetric trained medical staff in the health camps or transportation provided to take women with complications to the nearest health facility. The rate of adverse outcome following disaster was 22.2% (95% CI: 8.62% – 42.2%) amongst 27 women who did not evacuate as compare to 7.91% (95% CI: 5.03% – 11.8%) among 278 women who lived in relief camp study participants. There were 27 women who evacuated on pre-flood warning and had 0% rate of adverse outcome. Conclusion: We propose a model where CHWs will be used as frontline maternity care providers during any emergency or disaster situations in Pakistan. A separate "birthing station" should be mandatory in all district relief camps, managed by CHWs. Community midwives (CMW) would and the Lady Health Workers (LHW) would provide antenatal and postnatal care alongside, vaccination for pregnant women, neonates and children under five. There must be an ambulance facility for emergency obstetric cases and all district health facilities should have at least two medical staff identified and trained for emergency obstetric management. The District Health Department must provide clean birthing kits and regular and emergency contraceptives in the relief camps.

Keywords: natural disaster, maternity care model, rural, Pakistan, community health workers

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Authors: Ishita Singh

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Social-dimension of Climate Change is no longer peripheral to Science, Technology and Innovation (STI). Indeed, STI is being mobilized to address small farmers’ vulnerability and adaptation to Climate Change. The experiences from the cold desert of Leh (Ladakh) in North-West Himalaya illustrate the potential of STI to address the challenges of Climate Change and the needs of small farmers through the use of Artificial Glacier Techniques. Small farmers have a unique technique of water harvesting to augment irrigation, called “Artificial Glaciers” - an intricate network of water channels and dams along the upper slope of a valley that are located closer to villages and at lower altitudes than natural glaciers. It starts to melt much earlier and supplements additional irrigation to small farmers’ improving their livelihoods. Therefore, the issue of vulnerability, adaptive capacity and adaptation strategy needs to be analyzed in a local context and the communities as well as regions where people live. Leh (Ladakh) in North-West Himalaya provides a Case Study for exploring the ways in which adaptation to Climate Change is taking place at a community scale using Artificial Glacier Technology. With the above backdrop, an attempt has been made to analyze the rural poor households' vulnerability and adaptation practices to Climate Change using this technology, thereby drawing lessons on vulnerability-livelihood interactions in the cold desert of Leh (Ladakh) in North-West Himalaya, India. The study is based on primary data and information collected from 675 households confined to 27 villages of Leh (Ladakh) in North-West Himalaya, India. It reveals that 61.18% of the population is driving livelihoods from agriculture and allied activities. With increased irrigation potential due to the use of Artificial Glaciers, food security has been assured to 77.56% of households and health vulnerability has been reduced in 31% of households. Seasonal migration as a livelihood diversification mechanism has declined in nearly two-thirds of households, thereby improving livelihood strategies. Use of tactical adaptations by small farmers in response to persistent droughts, such as selling livestock, expanding agriculture lands, and use of relief cash and foods, have declined to 20.44%, 24.74% and 63% of households. However, these measures are unsustainable on a long-term basis. The role of policymakers and societal stakeholders becomes important in this context. To address livelihood challenges, the role of technology is critical in a multidisciplinary approach involving multilateral collaboration among different stakeholders. The presence of social entrepreneurs and new actors on the adaptation scene is necessary to bring forth adaptation measures. Better linkage between Science and Technology policies, together with other policies, should be encouraged. Better health care, access to safe drinking water, better sanitary conditions, and improved standards of education and infrastructure are effective measures to enhance a community’s adaptive capacity. However, social transfers for supporting climate adaptive capacity require significant amounts of additional investment. Developing institutional mechanisms for specific adaptation interventions can be one of the most effective ways of implementing a plan to enhance adaptation and build resilience.

Keywords: climate change, adaptation, livelihood, stakeholders

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Authors: N. K. Ambujam, V. Sudha

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Reservoirs are being contaminated all around the world with point source and Non-Point Source (NPS) pollution. The most common NPS pollutants are sediments and nutrients. Krishnagiri Reservoir (KR) has been chosen for the present case study, which is located in the tropical semi-arid climatic zone of Tamil Nadu, South India. It is the main source of surface water in Krishnagiri district to meet the freshwater demands. The reservoir has lost about 40% of its water holding capacity due to sedimentation over the period of 50 years. Hence, from the research and management perspective, there is a need for a sound knowledge on the spatial and seasonal variations of KR water quality. The present study encompasses the specific objectives as (i) to investigate the longitudinal heterogeneity and seasonal variations of physicochemical parameters, nutrients and biological characteristics of KR water and (ii) to examine the extent of degradation of water quality in KR. 15 sampling points were identified by uniform stratified method and a systematic monthly sampling strategy was selected due to high dynamic nature in its hydrological characteristics. The physicochemical parameters, major ions, nutrients and Chlorophyll a (Chl a) were analysed. Trophic status of KR was classified by using Carlson's Trophic State Index (TSI). All statistical analyses were performed by using Statistical Package for Social Sciences programme, version-16.0. Spatial maps were prepared for Chl a using Arc GIS. Observations in KR pointed out that electrical conductivity and major ions are highly variable factors as it receives inflow from the catchment with different land use activities. The study of major ions in KR exhibited different trends in their values and it could be concluded that as the monsoon progresses the major ions in the water decreases or water quality stabilizes. The inflow point of KR showed comparatively higher concentration of nutrients including nitrate, soluble reactive phosphorus (SRP), total phosphors (TP), total suspended phosphorus (TSP) and total dissolved phosphorus (TDP) during monsoon seasons. This evidently showed the input of significant amount of nutrients from the catchment side through agricultural runoff. High concentration of TDP and TSP at the lacustrine zone of the reservoir during summer season evidently revealed that there was a significant release of phosphorus from the bottom sediments. Carlson’s TSI of KR ranged between 81 and 92 during northeast monsoon and summer seasons. High and permanent Cyanobacterial bloom in KR could be mainly due to the internal loading of phosphorus from the bottom sediments. According to Carlson’s TSI classification Krishnagiri reservoir was ranked in the hyper-eutrophic category. This study provides necessary basic data on the spatio-temporal variations of water quality in KR and also proves the impact of point and NPS pollution from the catchment area. High TSI warrants a greater threat for the recovery of internal P loading and hyper-eutrophic condition of KR. Several expensive internal measures for the reduction of internal loading of P were introduced by many scientists. However, the outcome of the present research suggests for the innovative algae harvesting technique for the removal of sediment nutrients.

Keywords: NPS pollution, nutrients, hyper-eutrophication, krishnagiri reservoir

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Authors: Sarwan Dhir, Suma Basak, Dipika Parajulee

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Alfalfa is renowned for its nutritional and biopharmaceutical value as a perennial forage legume. However, establishing a rapid plant regeneration protocol using somatic embryogenesis and efficient transformation frequency are the crucial prerequisites for gene editing in alfalfa. This study was undertaken to establish and improve the protocol for somatic embryogenesis and subsequent plant regeneration. The experiments were conducted in response to natural sensitivity using various antibiotics such as cefotaxime, carbenicillin, gentamycin, hygromycin, and kanamycin. Using 3-week-old leaf tissue, somatic embryogenesis was initiated on Gamborg’s B5 basal (B5H) medium supplemented with 3% maltose, 0.9µM Kinetin, and 4.5µM 2,4-D. Embryogenic callus (EC) obtained from the B5H medium exhibited a high rate of somatic embryo formation (97.9%) after 3 weeks when the cultures were placed in the dark. Different developmental stages of somatic embryos and cotyledonary stages were then transferred to Murashige and Skoog’s (MS) basal medium under light, resulting in a 94% regeneration rate of plantlets. Our results indicate that leaf segments can grow (tolerate) up to 450 mg/L of cefotaxime and 400 mg/L of carbenicillin in the culture medium. However, the survival threshold for hygromycin at 12.5 mg/L, kanamycin at 250 mg/L, gentamycin at 50 mg/L, and timentin (300 mg/L). The experiment to improve the protocol for achieving efficient transient gene expression in alfalfa through genetic transformation with the Agrobacterium tumefaciens pCAMBIA1304 vector was also conducted. The vector contains two reporter genes such as β-glucuronidase (GUS) and green fluorescent protein (GFP), along with a selectable hygromycin B phosphotransferase gene (HPT), all driven under the CaMV 35s promoter. Various transformation parameters were optimized using 3-week-old in vitro-grown plantlets. The different parameters such as types of explant, leaf ages, preculture days, segment sizes, wounding types, bacterial concentrations, infection periods, co-cultivation periods, different concentrations of acetosyringone, silver nitrate, and calcium chloride were optimized for transient gene expression. The transient gene expression was confirmed via histochemical GUS and GFP visualization under fluorescent microscopy. The data were analyzed based on the semi-quantitative observation of the percentage and number of blue GUS spots on different days of agro-infection. The highest percentage of GUS positivity (76.2%) was observed in 3-week-old leaf segments wounded using a scalpel blade of 11 size- after 3 days of post-incubation at a bacterial concentration of 0.6, with 2 days of preculture, 30 min of bacterial-leaf segment co-cultivation, with the addition of 150 µM acetosyringone, 4 mM calcium chloride, and 75 µM silver nitrate. Our results suggest that various factors influence T-DNA delivery in the Agrobacterium-mediated transformation of alfalfa. The stable gene expression in the putative transgenic tissue was confirmed using PCR amplification of both marker genes, indicating that gene expression in explants was not solely due to Agrobacterium, but also from transformed cells. The improved protocol could be used for generating transgenic alfalfa plants using genome editing techniques such as CRISPR/Cas9.

Keywords: Medicago sativa l. (Alfalfa), agrobacterium tumefaciens, β-glucuronidase, green fluorescent protein, transient gene

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Authors: Nurkhairul Bariyah Baharun, Afzan Adam, Reena Rahayu Md Zin

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Tumor microenvironment (TME) in breast cancer is mainly composed of cancer cells, immune cells, and stromal cells. The interaction between cancer cells and their microenvironment plays an important role in tumor development, progression, and treatment response. The TME in breast cancer includes tumor-infiltrating lymphocytes (TILs) that are implicated in killing tumor cells. TILs can be found in tumor stroma (sTILs) and within the tumor (iTILs). TILs in triple negative breast cancer (TNBC) have been demonstrated to have prognostic and potentially predictive value. The international Immune-Oncology Biomarker Working Group (TIL-WG) had developed a guideline focus on the assessment of sTILs using hematoxylin and eosin (H&E)-stained slides. According to the guideline, the pathologists use “eye balling” method on the H&E stained- slide for sTILs assessment. This method has low precision, poor interobserver reproducibility, and is time-consuming for a comprehensive evaluation, besides only counted sTILs in their assessment. The TIL-WG has therefore recommended that any algorithm for computational assessment of TILs utilizing the guidelines provided to overcome the limitations of manual assessment, thus providing highly accurate and reliable TILs detection and classification for reproducible and quantitative measurement. This study is carried out to develop a TNBC digital whole slide image (WSI) dataset from H&E-stained slides and IHC (CD4+ and CD8+) stained slides. TNBC cases were retrieved from the database of the Department of Pathology, Hospital Canselor Tuanku Muhriz (HCTM). TNBC cases diagnosed between the year 2010 and 2021 with no history of other cancer and available block tissue were included in the study (n=58). Tissue blocks were sectioned approximately 4 µm for H&E and IHC stain. The H&E staining was performed according to a well-established protocol. Indirect IHC stain was also performed on the tissue sections using protocol from Diagnostic BioSystems PolyVue™ Plus Kit, USA. The slides were stained with rabbit monoclonal, CD8 antibody (SP16) and Rabbit monoclonal, CD4 antibody (EP204). The selected and quality-checked slides were then scanned using a high-resolution whole slide scanner (Pannoramic DESK II DW- slide scanner) to digitalize the tissue image with a pixel resolution of 20x magnification. A manual TILs (sTILs and iTILs) assessment was then carried out by the appointed pathologist (2 pathologists) for manual TILs scoring from the digital WSIs following the guideline developed by TIL-WG 2014, and the result displayed as the percentage of sTILs and iTILs per mm² stromal and tumour area on the tissue. Following this, we aimed to develop an automated digital image scoring framework that incorporates key elements of manual guidelines (including both sTILs and iTILs) using manually annotated data for robust and objective quantification of TILs in TNBC. From the study, we have developed a digital dataset of TNBC H&E and IHC (CD4+ and CD8+) stained slides. We hope that an automated based scoring method can provide quantitative and interpretable TILs scoring, which correlates with the manual pathologist-derived sTILs and iTILs scoring and thus has potential prognostic implications.

Keywords: automated quantification, digital pathology, triple negative breast cancer, tumour infiltrating lymphocytes

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Authors: Fadi Al Khatib, Raouf Mbarki, Malek Adouni

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In various sport and recreational activities, the patellofemoral joint undergoes large forces and moments while accommodating the significant knee joint movement. In doing so, this joint is commonly the source of anterior knee pain related to instability in normal patellar tracking and excessive pressure syndrome. One well-observed explanation of the instability of the normal patellar tracking is the patellofemoral ligaments and patellar tendon damage. Improved knowledge of the damage mechanism mediating ligaments and tendon injuries can be a great help not only in rehabilitation and prevention procedures but also in the design of better reconstruction systems in the management of knee joint disorders. This damage mechanism, specifically due to excessive mechanical loading, has been linked to the micro level of the fibred structure precisely to the tropocollagen molecules and their connection density. We argue defining a clear frame starting from the bottom (micro level) to up (macro level) in the hierarchies of the soft tissue may elucidate the essential underpinning on the state of the ligaments damage. To do so, in this study a multiscale fibril reinforced hyper elastoplastic Finite Element model that accounts for the synergy between molecular and continuum syntheses was developed to determine the short-term stresses/strains patellofemoral ligaments and tendon response. The plasticity of the proposed model is associated only with the uniaxial deformation of the collagen fibril. The yield strength of the fibril is a function of the cross-link density between tropocollagen molecules, defined here by a density function. This function obtained through a Coarse-graining procedure linking nanoscale collagen features and the tissue level materials properties using molecular dynamics simulations. The hierarchies of the soft tissues were implemented using the rule of mixtures. Thereafter, the model was calibrated using a statistical calibration procedure. The model then implemented into a real structure of patellofemoral ligaments and patellar tendon (OpenKnee) and simulated under realistic loading conditions. With the calibrated material parameters the calculated axial stress lies well with the experimental measurement with a coefficient of determination (R2) equal to 0.91 and 0.92 for the patellofemoral ligaments and the patellar tendon respectively. The ‘best’ prediction of the yielding strength and strain as compared with the reported experimental data yielded when the cross-link density between the tropocollagen molecule of the fibril equal to 5.5 ± 0.5 (patellofemoral ligaments) and 12 (patellar tendon). Damage initiation of the patellofemoral ligaments was located at the femoral insertions while the damage of the patellar tendon happened in the middle of the structure. These predicted finding showed a meaningful correlation between the cross-link density of the tropocollagen molecules and the stiffness of the connective tissues of the extensor mechanism. Also, damage initiation and propagation were documented with this model, which were in satisfactory agreement with earlier observation. To the best of our knowledge, this is the first attempt to model ligaments from the bottom up, predicted depending to the tropocollagen cross-link density. This approach appears more meaningful towards a realistic simulation of a damaging process or repair attempt compared with certain published studies.

Keywords: tropocollagen, multiscale model, fibrils, knee ligaments

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