Search results for: human knowledge
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 14608

Search results for: human knowledge

628 Comparison of Gestational Diabetes Influence on the Ultrastructure of Rectus Abdominis Muscle in Women and Rats

Authors: Giovana Vesentini, Fernanda Piculo, Gabriela Marini, Debora Damasceno, Angelica Barbosa, Selma Martheus, Marilza Rudge

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Problem statement: Skeletal muscle is highly adaptable, muscle fiber composition and size can respond to a variety of stimuli, such physiologic, as pregnancy, and metabolic abnormalities, as Diabetes mellitus. This study aimed to analyze the effects of pregnancy-associated diabetes on the rectus abdominis muscle (RA), and to compare this changes in rats and women. Methods: Female Wistar rats were maintained under controlled conditions and distributed in Pregnant (P) and Long-term mild pregnant diabetic (LTMd) (n=3 r/group). Diabetes in rats was induced by streptozotocin (100mg/Kg, sc) on the first day of life, for a hyperglycemic state between 120-300 mg/dL in adult life. Female rats were mated overnight, at day 21 of pregnancy were anesthetized, and killed for the harvesting of maternal RA. Pregnant women who attended the Diabetes Prenatal Care Clinic of Botucatu Medical School were distributed in Pregnant non-diabetic (Pnd) and Gestational Diabetic (GDM) (n=3 w/group). The diagnosis of GDM was established according to ADA’s criteria (2016). The harvesting of RA was during the cesarean section. Transversal cross-sections of the RA of both women and rats were analyzed by transmission electron microscopy. All procedures were approved by the Ethics Committee on Animal Experiments of the Botucatu Medical School (Protocol Number 1003/2013) and by the Botucatu Medical School Ethical Committee for Human Research in Medical Sciences (CAAE: 41570815.0.0000.5411). Results: The photomicrographs of the RA of rats revealed disorganized Z lines, thinning sarcomeres, and a usual quantity of intermyofibrillar mitochondria in the P group. The LTMd group showed swollen sarcoplasmic reticulum, dilated T tubes and areas with sarcomere disruption. The ultrastructural analysis of Pnd non-diabetic women in the RA showed well-organized myofibrils forming intact sarcomeres, organized Z lines and a normal distribution of intermyofibrillar mitochondria. The GDM group revealed increase in intermyofibrillar mitochondria, areas with sarcomere disruption and increased lipid droplets. Conclusion: Pregnancy and diabetes induce adaptations in the ultrastructure of the rectus abdominis muscle for both women and rats, changing the architectural design of these tissues. However, in rats these changes are more severe maybe because, besides the high blood glucose levels, the quadrupedal animal may suffer an excessive mechanical tension during pregnancy by gravity. Probably, these findings may suggest that these alterations are a risk factor that contributes to the development of muscle dysfunction in women with GDM and may motivate treatment strategies in these patients.

Keywords: gestational diabetes, muscle dysfunction, pregnancy, rectus abdominis

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627 Strengths Profiling: An Alternative Approach to Assessing Character Strengths Based on Personal Construct Psychology

Authors: Sam J. Cooley, Mary L. Quinton, Benjamin J. Parry, Mark J. G. Holland, Richard J. Whiting, Jennifer Cumming

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Practitioners draw attention to people’s character strengths to promote empowerment and well-being. This paper explores the possibility that existing approaches for assessing character strengths (e.g., the Values in Action survey; VIA-IS) could be even more autonomy supportive and empowering when combined with strengths profiling, an ideographic tool informed by personal construct theory (PCT). A PCT approach ensures that: (1) knowledge is co-created (i.e., the practitioner is not seen as the ‘expert’ who leads the process); (2) individuals are not required to ‘fit’ within a prescribed list of characteristics; and (3) individuals are free to use their own terminology and interpretations. A combined Strengths Profiling and VIA approach was used in a sample of homeless youth (aged 16-25) who are commonly perceived as ‘hard-to-engage’ through traditional forms of assessment. Strengths Profiling was completed face-to-face in small groups. Participants (N = 116) began by listing a variety of personally meaningful characteristics. Participants gave each characteristic a score out of ten for how important it was to them (1 = not so important; 10 = very important), their ideal competency, and their current competency (1 = poor; 10 = excellent). A discrepancy score was calculated for each characteristic (discrepancy score = ideal score - current score x importance), whereby a lower discrepancy score indicated greater satisfaction. Strengths Profiling was used at the beginning and end of a 10-week positive youth development programme. Experiences were captured through video diary room entries made by participants and through reflective notes taken by the facilitators. Participants were also asked to complete a pre-and post-programme questionnaire, measuring perceptions of well-being, self-worth, and resilience. All of the young people who attended the strengths profiling session agreed to complete a profile, and the majority became highly engaged in the process. Strengths profiling was found to be an autonomy supportive and empowering experience, with each participant identifying an average of 10 character strengths (M = 10.27, SD = 3.23). In total, 215 different character strengths were identified, each with varying terms and definitions used, which differed greatly between participants and demonstrated the value in soliciting personal constructs. Using the participants’ definitions, 98% of characteristics were categorized deductively into the VIA framework. Bravery, perseverance, and hope were the character strengths that featured most, whilst temperance and courage received the highest discrepancy scores. Discrepancy scores were negatively correlated with well-being, self-worth, and resilience, and meaningful improvements were recorded following the intervention. These findings support the use of strengths profiling as a theoretically-driven and novel way to engage disadvantaged youth in identifying and monitoring character strengths. When young people are given the freedom to express their own characteristics, the resulting terminologies extend beyond the language used in existing frameworks. This added freedom and control over the process of strengths identification encouraged youth to take ownership over their profiles and apply their strengths. In addition, the ability to transform characteristics post hoc into the VIA framework means that strengths profiling can be used to explore aggregated/nomothetic hypotheses, whilst still benefiting from its ideographic roots.

Keywords: ideographic, nomothetic, positive youth development, VIA-IS, assessment, homeless youth

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626 Recovery of Food Waste: Production of Dog Food

Authors: K. Nazan Turhan, Tuğçe Ersan

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The population of the world is approximately 8 billion, and it increases uncontrollably and irrepressibly, leading to an increase in consumption. This situation causes crucial problems, and food waste is one of these. The Food and Agriculture Organization of the United Nations (FAO) defines food waste as the discarding or alternative utilization of food that is safe and nutritious for the consumption of humans along the entire food supply chain, from primary production to end household consumer level. In addition, according to the estimation of FAO, one-third of all food produced for human consumption is lost or wasted worldwide every year. Wasting food endangers natural resources and causes hunger. For instance, excessive amounts of food waste cause greenhouse gas emissions, contributing to global warming. Therefore, waste management has been gaining significance in the last few decades at both local and global levels due to the expected scarcity of resources for the increasing population of the world. There are several ways to recover food waste. According to the United States Environmental Protection Agency’s Food Recovery Hierarchy, food waste recovery ways are source reduction, feeding hungry people, feeding animals, industrial uses, composting, and landfill/incineration from the most preferred to the least preferred, respectively. Bioethanol, biodiesel, biogas, agricultural fertilizer and animal feed can be obtained from food waste that is generated by different food industries. In this project, feeding animals was selected as a food waste recovery method and food waste of a plant was used to provide ingredient uniformity. Grasshoppers were used as a protein source. In other words, the project was performed to develop a dog food product by recovery of the plant’s food waste after following some steps. The collected food waste and purchased grasshoppers were sterilized, dried and pulverized. Then, they were all mixed with 60 g agar-agar solution (4%w/v). 3 different aromas were added, separately to the samples to enhance flavour quality. Since there are differences in the required amounts of different species of dogs, fulfilling all nutritional needs is one of the problems. In other words, there is a wide range of nutritional needs in terms of carbohydrates, protein, fat, sodium, calcium, and so on. Furthermore, the requirements differ depending on age, gender, weight, height, and species. Therefore, the product that was developed contains average amounts of each substance so as not to cause any deficiency or surplus. On the other hand, it contains more protein than similar products in the market. The product was evaluated in terms of contamination and nutritional content. For contamination risk, detection of E. coli and Salmonella experiments were performed, and the results were negative. For the nutritional value test, protein content analysis was done. The protein contents of different samples vary between 33.68% and 26.07%. In addition, water activity analysis was performed, and the water activity (aw) values of different samples ranged between 0.2456 and 0.4145.

Keywords: food waste, dog food, animal nutrition, food waste recovery

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625 A High Amylose-Content and High-Yielding Elite Line Is Favorable to Cook 'Nanhan' (Semi-Soft Rice) for Nursing Care Food Particularly for Serving Aged Persons

Authors: M. Kamimukai, M. Bhattarai, B. B. Rana, K. Maeda, H. B. Kc, T. Kawano, M. Murai

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Most of the aged people older than 70 have difficulty in chewing and swallowing more or less. According to magnitude of this difficulty, gruel, “nanhan” (semi-soft rice) and ordinary cooked rice are served in general, particularly in sanatoriums and homes for old people in Japan. Nanhan is the name of a cooked rice used in Japan, having softness intermediate between gruel and ordinary cooked rice, which is boiled with intermediate amount of water between those of the latter two kinds of cooked rice. In the present study, nanhan was made in the rate of 240g of water to 100g of milled rice with an electric rice cooker. Murai developed a high amylose-content and high-yielding elite line ‘Murai 79’. Sensory eating-quality test was performed for nanhan and ordinary cooked rice of Murai 79 and the standard variety ‘Hinohikari’ which is a high eating-quality variety representative in southern Japan. Panelists (6 to 14 persons) scored each cooked rice in six items viz. taste, stickiness, hardness, flavor, external appearance and overall evaluation. Grading (-3 ~ +3) in each trait was performed, regarding the value of the standard variety Hinohikari as 0. Paddy rice produced in a farmer’s field in 2013 and 2014 and in an experimental field of Kochi University in 2015 and 2016 were used for the sensory test. According to results of the sensory eating-quality test for nanhan, Murai 79 is higher in overall evaluation than Hinohikari in the four years. The former was less sticky than the latter in the four years, but the former was statistically significantly harder than the latter throughout the four years. In external appearance, the former was significantly higher than the latter in the four years. In the taste, the former was significantly higher than the latter in 2014, but significant difference was not noticed between them in the other three years. There were no significant differences throughout the four years in flavor. Regarding amylose content, Murai 79 is higher by 3.7 and 5.7% than Hinohikari in 2015 and 2016, respectively. As for protein content, Murai 79 was higher than Hinohikari in 2015, but the former was lower than the latter in 2016. Consequently, the nanhan of Murai 79 was harder and less sticky, keeping the shape of grains as compared with that of Hinohikari, which may be due to its higher amylose content. Hence, the nanhan of Murai 79 may be recognized as grains more easily in a human mouth, which could make easier the continuous performance of mastication and deglutition particularly in aged persons. Regarding ordinary cooked rice, Murai 79 was similar to or higher in both overall evaluation and external appearance as compared with Hinohikari, despite its higher hardness and lower stickiness. Additionally, Murai 79 had brown-rice yield of 1.55 times as compared with Hinohikari, suggesting that it would enable to supply inexpensive rice for making nanhan with high quality particularly for aged people in Japan.

Keywords: high-amylose content, high-yielding rice line, nanhan, nursing care food, sensory eating quality test

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624 Differentially Expressed Protein Biomarkers in Early and Advanced Stage Young Triple-Negative Breast Cancer Patients

Authors: Shamim Mushtaq, Moazzam Shahid

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Breast cancer (BC) claims the lives of half a million women every year and is the most common cause of death in the developing world. In 2019, it was estimated that BC alone accounts for 15% of all cancer deaths in younger women (aged < 45 years old) with advanced-stage lung metastasis. According to the World Health Organization & International Union against Cancer, in Asia, a high number of cancer-related deaths will be observed in 2020, whereas the burden will be reduced in Western countries due to awareness about the disease, better health facilities and advanced treatments. In the last 15 years, it has been reported that the incidence of BC has increased by 1.1% among Asian compared to the US population from 2003 to 2012. To date, several BC biological subtypes have been reported so far, which are associated with different treatment responses. The heterogeneity and diversity of BC reflected these different subtypes, including Luminal A (23.7% prevalence) and B (38.8% prevalence) that have pathological estrogen receptor (ER+)-positive tumors, the human epidermal growth factor receptor 2 (HER2) (11.2% prevalence) and triple-negative breast cancer (TNBC) (25% prevalence). According to Shaukat Khanum Memorial Cancer Hospital and Research Centre – Pakistan, ten years of data showed that among 636 BC patients, 30.5% had TNBC who were <40 years of age, which is an extremely alarming situation. Therefore, there is a dire need to explore and develop therapeutic targets for the treatment of early TNBC. Since the last decade, unfortunately, there has been little success in understanding the complexity of TNBC and in discovering new biological therapeutic targets. However, conventional chemotherapy is the only choice of treatment for TNBC patients. Many investigators revealed advances in multi-omics (multiple "omes", e.g., genome, proteome, transcriptome, epigenome, and microbiome) which were later identified as actionable targets and increased prevalence in TNBC patients. However, various drugs have been identified so far which are related to a particular diagnostic and prognostic biomarker. For example, Epidermal growth factor receptor ( EGFR or ErbB-1), HER-2/neu (ErbB-2), HER-3 (ErbB-3), and HER-4 (ErbB-4). Protein Transglin-2 (TAGLN 2 ) and Profilins-1 (Pfn-1 ) are the ubiquitously expressed large family of proteins present in all eukaryotes, enabling actin cytoskeletal reorganization. It is known that the oncogenic transformation of cells is accompanied by alteration in the actin cytoskeleton. There are causal connections between altered expression of actin cytoskeletal regulators and cancer progression. Our case-control study identified TAGLN-2 and Pfn-1 proteins in TNBC blood by mass spectrometry. Both TAGLN-2 and Pfn-1 proteins are differentially expressed in early and advanced stages of TNBS patients, which could be potential predictors or therapeutic targets for TNBC.

Keywords: TNBC, blood biomarkers, mass spectrometry, qPCR, ELISA

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623 Basic Education Curriculum in South- South Nigeria: Challenges and Opportunities of Quality Contents in the Second Language Learning

Authors: Catherine Alex Agbor

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The modern Nigerian society is dynamic, divided in zones based on economic, political and educational resources often shared across the zones. The Six Geopolitical Zones in Nigeria is a major division in modern Nigeria, created during the regime of president Ibrahim Badamasi Babangida. They are North Central, North East, North West, South East, South South and South West. However, the zone used in this study is known as former South-Eastern State of Akwa-Ibom State and Cross-River State; former Rivers State of Bayelsa State and Rivers State; and former Mid-Western Region, Nigeria of Delta State and Edo State. Many reforms have taken place overtime, particularly in the education sector. Education is constantly presenting new ideas and innovative approaches which act to facilitate the rapid exchange of knowledge and provide quality basic education for learners. The Federal Government of Nigeria in accordance with its National Council on Education directed the Nigerian Educational Research and Development Council to restructure its basic education curriculum with the hope to enable the nation meet national and global developmental goals. One of the goals of the 9-year Basic Education Programme is developing in the entire citizenry a strong consciousness for education and a strong commitment to its vigorous promotion. Another is ensuring the acquisition of appropriate levels of literacy, numeracy, manipulative, communicative and life-skills as well as the ethical, moral and civic values for laying a solid foundation for lifelong learning. Therefore, this article at the introductory stage is aimed to describe some key issues in Nigeria’s experience in the basic education curriculum. In this study, particular attention is paid to this very recent educational policy of the Nigerian government known as Universal Basic Education, its challenges and what can be done to make the policy achieve its desired objectives. It progresses to analyze modern requirements for second language teaching; and presents the challenges of second language teaching in Nigeria. Finally, it reports a study which investigated special efforts for appropriate achievement of quality education in language classroom in the south-south zone of Nigeria. One fundamental research question was posed on what educational practices can contribute to current understanding of the structure of language curriculum. More explicitly, the study was designed to analyze the extent to which quality content contributes to current understanding of the structure of school curriculum in the zone. Otherwise stated, it investigated how student-centred educational practices impact on their learning of French language. One hundred and eighty (180) participants (teachers) were purposefully sampled for the study. Qualitative technique was used to elicit information from participants. The qualitative method used was Focus Group Discussion (FGD). Participants were divided into six groups comprising of 30 teachers from each zone. Group discussions were based mainly on curriculum contents and practices. Information from participants revealed that the curriculum content, among others is inadequate and should be re-examined. Recommendations were proffered as a panacea to concrete implementation of the basic education in Nigeria.

Keywords: basic education, quality contents, second language, south-south states

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622 The Development of Wind Energy and Its Social Acceptance: The Role of Income Received by Wind Farm Owners, the Case of Galicia, Northwest Spain

Authors: X. Simon, D. Copena, M. Montero

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The last decades have witnessed a significant increase in renewable energy, especially wind energy, to achieve sustainable development. Specialized literature in this field has carried out interesting case studies to extensively analyze both the environmental benefits of this energy and its social acceptance. However, to the best of our knowledge, work to date makes no analysis of the role of private owners of lands with wind potential within a broader territory of strong wind implantation, nor does it estimate their economic incomes relating them to social acceptance. This work fills this gap by focusing on Galicia, territory housing over 4,000 wind turbines and almost 3,400 MW of power. The main difficulty in getting this financial information is that it is classified, not public. We develop methodological techniques (semi- structured interviews and work groups), inserted within the Participatory Research, to overcome this important obstacle. In this manner, the work directly compiles qualitative and quantitative information on the processes as well as the economic results derived from implementing wind energy in Galicia. During the field work, we held 106 semi-structured interviews and 32 workshops with owners of lands occupied by wind farms. The compiled information made it possible to create the socioeconomic database on wind energy in Galicia (SDWEG). This database collects a diversity of quantitative and qualitative information and contains economic information on the income received by the owners of lands occupied by wind farms. In the Galician case, regulatory framework prevented local participation under the community wind farm formula. The possibility of local participation in the new energy model narrowed down to companies wanting to install a wind farm and demanding land occupation. The economic mechanism of local participation begins here, thus explaining the level of acceptance of wind farms. Land owners can receive significant income given that these payments constitute an important source of economic resources, favor local economic activity, allow rural areas to develop productive dynamism projects and improve the standard of living of rural inhabitants. This work estimates that land owners in Galicia perceive about 10 million euros per year in total wind revenues. This represents between 1% and 2% of total wind farm invoicing. On the other hand, relative revenues (Euros per MW), far from the amounts reached in other spaces, show enormous payment variability. This signals the absence of a regulated market, the predominance of partial agreements, and the existence of asymmetric positions between owners and developers. Sustainable development requires the replacement of conventional technologies by low environmental impact technologies, especially those that emit less CO₂. However, this new paradigm also requires rural owners to participate in the income derived from the structural transformation processes linked to sustainable development. This paper demonstrates that regulatory framework may contribute to increasing sustainable technologies with high social acceptance without relevant local economic participation.

Keywords: regulatory framework, social acceptance, sustainable development, wind energy, wind income for landowners

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621 Municipalities as Enablers of Citizen-Led Urban Initiatives: Possibilities and Constraints

Authors: Rosa Nadine Danenberg

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In recent years, bottom-up urban development has started growing as an alternative to conventional top-down planning. In large proportions, citizens and communities initiate small-scale interventions; suddenly seeming to form a trend. As a result, more and more cities are witnessing not only the growth of but also an interest in these initiatives, as they bear the potential to reshape urban spaces. Such alternative city-making efforts cause new dynamics in urban governance, with inevitable consequences for the controlled city planning and its administration. The emergence of enabling relationships between top-down and bottom-up actors signals an increasingly common urban practice. Various case studies show that an enabling relationship is possible, yet, how it can be optimally realized stays rather underexamined. Therefore, the seemingly growing worldwide phenomenon of ‘municipal bottom-up urban development’ necessitates an adequate governance structure. As such, the aim of this research is to contribute knowledge to how municipalities can enable citizen-led urban initiatives from a governance innovation perspective. Empirical case-study research in Stockholm and Istanbul, derived from interviews with founders of four citizen-led urban initiatives and one municipal representative in each city, provided valuable insights to possibilities and constraints for enabling practices. On the one hand, diverging outcomes emphasize the extreme oppositional features of both cases (Stockholm and Istanbul). Firstly, both cities’ characteristics are drastically different. Secondly, the ideologies and motifs for the initiatives to emerge vary widely. Thirdly, the major constraints for citizen-led urban initiatives to relate to the municipality are considerably different. Two types of municipality’s organizational structures produce different underlying mechanisms which demonstrate the constraints. The first municipal organizational structure is steered by bureaucracy (Stockholm). It produces an administrative division that brings up constraints such as the lack of responsibility, transparency and continuity by municipal representatives. The second structure is dominated by municipal politics and governmental hierarchy (Istanbul). It produces informality, lack of transparency and a fragmented civil society. In order to cope with the constraints produced by both types of organizational structures, the initiatives have adjusted their organization to the municipality’s underlying structures. On the other hand, this paper has in fact also come to a rather unifying conclusion. Interestingly, the suggested possibilities for an enabling relationship underline converging new urban governance arrangements. This could imply that for the two varying types of municipality’s organizational structures there is an accurate governance structure. Namely, the combination of a neighborhood council with a municipal guide, with allowance for the initiatives to adopt a politicizing attitude is found as coinciding. Especially its combination appears key to redeem varying constraints. A municipal guide steers the initiatives through bureaucratic struggles, is supported by coproduction methods, while it balances out municipal politics. Next, a neighborhood council, that is politically neutral and run by local citizens, can function as an umbrella for citizen-led urban initiatives. What is crucial is that it should cater for a more entangled relationship between municipalities and initiatives with enhanced involvement of the initiatives in decision-making processes and limited involvement of prevailing constraints pointed out in this research.

Keywords: bottom-up urban development, governance innovation, Istanbul, Stockholm

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620 Ibrutinib and the Potential Risk of Cardiac Failure: A Review of Pharmacovigilance Data

Authors: Abdulaziz Alakeel, Roaa Alamri, Abdulrahman Alomair, Mohammed Fouda

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Introduction: Ibrutinib is a selective, potent, and irreversible small-molecule inhibitor of Bruton's tyrosine kinase (BTK). It forms a covalent bond with a cysteine residue (CYS-481) at the active site of Btk, leading to inhibition of Btk enzymatic activity. The drug is indicated to treat certain type of cancers such as mantle cell lymphoma (MCL), chronic lymphocytic leukaemia and Waldenström's macroglobulinaemia (WM). Cardiac failure is a condition referred to inability of heart muscle to pump adequate blood to human body organs. There are multiple types of cardiac failure including left and right-sided heart failure, systolic and diastolic heart failures. The aim of this review is to evaluate the risk of cardiac failure associated with the use of ibrutinib and to suggest regulatory recommendations if required. Methodology: Signal Detection team at the National Pharmacovigilance Center (NPC) of Saudi Food and Drug Authority (SFDA) performed a comprehensive signal review using its national database as well as the World Health Organization (WHO) database (VigiBase), to retrieve related information for assessing the causality between cardiac failure and ibrutinib. We used the WHO- Uppsala Monitoring Centre (UMC) criteria as standard for assessing the causality of the reported cases. Results: Case Review: The number of resulted cases for the combined drug/adverse drug reaction are 212 global ICSRs as of July 2020. The reviewers have selected and assessed the causality for the well-documented ICSRs with completeness scores of 0.9 and above (35 ICSRs); the value 1.0 presents the highest score for best-written ICSRs. Among the reviewed cases, more than half of them provides supportive association (four probable and 15 possible cases). Data Mining: The disproportionality of the observed and the expected reporting rate for drug/adverse drug reaction pair is estimated using information component (IC), a tool developed by WHO-UMC to measure the reporting ratio. Positive IC reflects higher statistical association while negative values indicates less statistical association, considering the null value equal to zero. The results of (IC=1.5) revealed a positive statistical association for the drug/ADR combination, which means “Ibrutinib” with “Cardiac Failure” have been observed more than expected when compared to other medications available in WHO database. Conclusion: Health regulators and health care professionals must be aware for the potential risk of cardiac failure associated with ibrutinib and the monitoring of any signs or symptoms in treated patients is essential. The weighted cumulative evidences identified from causality assessment of the reported cases and data mining are sufficient to support a causal association between ibrutinib and cardiac failure.

Keywords: cardiac failure, drug safety, ibrutinib, pharmacovigilance, signal detection

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619 Possibilities of Psychodiagnostics in the Context of Highly Challenging Situations in Military Leadership

Authors: Markéta Chmelíková, David Ullrich, Iva Burešová

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The paper maps the possibilities and limits of diagnosing selected personality and performance characteristics of military leadership and psychology students in the context of coping with challenging situations. Individuals vary greatly inter-individually in their ability to effectively manage extreme situations, yet existing diagnostic tools are often criticized mainly for their low predictive power. Nowadays, every modern army focuses primarily on the systematic minimization of potential risks, including the prediction of desirable forms of behavior and the performance of military commanders. The context of military leadership is well known for its life-threatening nature. Therefore, it is crucial to research stress load in the specific context of military leadership for the purpose of possible anticipation of human failure in managing extreme situations of military leadership. The aim of the submitted pilot study, using an experiment of 24 hours duration, is to verify the possibilities of a specific combination of psychodiagnostic to predict people who possess suitable equipment for coping with increased stress load. In our pilot study, we conducted an experiment of 24 hours duration with an experimental group (N=13) in the bomb shelter and a control group (N=11) in a classroom. Both groups were represented by military leadership students (N=11) and psychology students (N=13). Both groups were equalized in terms of study type and gender. Participants were administered the following test battery of personality characteristics: Big Five Inventory 2 (BFI-2), Short Dark Triad (SD-3), Emotion Regulation Questionnaire (ERQ), Fatigue Severity Scale (FSS), and Impulsive Behavior Scale (UPPS-P). This test battery was administered only once at the beginning of the experiment. Along with this, they were administered a test battery consisting of the Test of Attention (d2) and the Bourdon test four times overall with 6 hours ranges. To better simulate an extreme situation – we tried to induce sleep deprivation - participants were required to try not to fall asleep throughout the experiment. Despite the assumption that a stay in an underground bomb shelter will manifest in impaired cognitive performance, this expectation has been significantly confirmed in only one measurement, which can be interpreted as marginal in the context of multiple testing. This finding is a fundamental insight into the issue of stress management in extreme situations, which is crucial for effective military leadership. The results suggest that a 24-hour stay in a shelter, together with sleep deprivation, does not seem to simulate sufficient stress for an individual, which would be reflected in the level of cognitive performance. In the context of these findings, it would be interesting in future to extend the diagnostic battery with physiological indicators of stress, such as: heart rate, stress score, physical stress, mental stress ect.

Keywords: bomb shelter, extreme situation, military leadership, psychodiagnostic

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618 From Isolation to Integration: A Biophilic Design Approach for Enhancing Inhabitants’ Well-being in Urban Residential Spaces in Dhaka

Authors: Maliha Afroz Nitu, Shahreen Mukashafat Semontee

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The concept of biophilic design has emerged as a transformative approach to restore the intrinsic connection between people and nature, an innate bond disrupted by urbanization and industrialization. As urbanization progresses, it is crucial to raise awareness about these issues in order to ensure people can live and work in healthy environments that enhance well-being. Dhaka, the capital of Bangladesh, faces challenges arising from unplanned urban expansion, leading to a notable disconnect between city dwellers and their natural surroundings, a problem prevalent in rapidly developing megacities. Significant interdisciplinary research consistently shows that connecting indoor and outdoor spaces can improve mental and physical well-being by rekindling a connection with the natural world. However, there is a significant lack of study on the implementation of biophilic design principles in the built environment to tackle these problems, despite the well-documented advantages. The Palashi Government Staff Quarter, a 3.8-acre housing area for government staff with around 1,000 residents in Dhaka, has been selected as a case study. The main goal is to create and implement biophilic design solutions to address social, environmental, and health issues while also enhancing the built environment. A methodology applicable to improving biophilic design is developed according to the needs of the residents. This research uses a comprehensive approach, including site inspections and structured and semi-structured interviews with residents to gather qualitative data on their experiences and needs. A total of ten identical six-story buildings have been surveyed, with varying resident responses providing insight into their different perspectives. Based on these findings, the study proposes alternative design strategies that integrate biophilic elements such as daylight, air, plants, and water into buildings through windows, skylights, clerestories, green walls, vegetation, and constructed water bodies. The objective of these strategies is to improve the built environment that restores the existing disconnection between humans and nature. Comparative analyses of the current and proposed scenarios demonstrate substantial upgrades in the built environment, as well as major improvements in the physical and psychological well-being of residents. Although this research focuses on a particular government housing, the findings can be applied to other residential areas in Dhaka and similar urban environments. The study highlights the importance of biophilic design in housing and provides recommendations for policymakers and architects to improve living conditions by integrating nature into urban settings.

Keywords: biophilic design, residential, built environment, human nature connection, urban, Dhaka

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617 Human Immuno-Deficiency Virus Co-Infection with Hepatitis B Virus and Baseline Cd4+ T Cell Count among Patients Attending a Tertiary Care Hospital, Nepal

Authors: Soma Kanta Baral

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Background: Since 1981, when the first AIDS case was reported, worldwide, more than 34 million people have been infected with HIV. Almost 95 percent of the people infected with HIV live in developing countries. As HBV & HIV share similar routes of transmission by sexual intercourse or drug use by parenteral injection, co-infection is common. Because of the limited access to healthcare & HIV treatment in developing countries, HIV-infected individuals are present late for care. Enumeration of CD4+ T cell count at the time of diagnosis has been useful to initiate the therapy in HIV infected individuals. The baseline CD4+ T cell count shows high immunological variability among patients. Methods: This prospective study was done in the serology section of the Department of Microbiology over a period of one year from august 2012 to July 2013. A total of 13037 individuals subjected for HIV test were included in the study comprising of 4982 males & 8055 females. Blood sample was collected by vein puncture aseptically with standard operational procedure in clean & dry test-tube. All blood samples were screened for HIV as described by WHO algorithm by Immuno-chromatography rapid kits. Further confirmation was done by biokit ELISA method as per the manufacturer’s guidelines. After informed consent, HIV positive individuals were screened for HBsAg by Immuno-chromatography rapid kits (Hepacard). Further confirmation was done by biokit ELISA method as per the manufacturer’s guidelines. EDTA blood samples were collected from the HIV sero-positive individuals for baseline CD4+ T count. Then, CD4+ T cells count was determined by using FACS Calibur Flow Cytometer (BD). Results: Among 13037 individuals screened for HIV, 104 (0.8%) were found to be infected comprising of 69(66.34%) males & 35 (33.65%) females. The study showed that the high infection was noted in housewives (28.7%), active age group (30.76%), rural area (56.7%) & in heterosexual route (80.9%) of transmission. Out of total HIV infected individuals, distribution of HBV co-infection was found to be 6(5.7%). All co- infected individuals were married, male, above the age of 25 years & heterosexual route of transmission. Baseline CD4+ T cell count of HIV infected patient was found higher (mean CD4+ T cell count; 283cells/cu.mm) than HBV co-infected patients (mean CD4+ T cell count; 91 cells/cu.mm). Majority (77.2%) of HIV infected & all co-infected individuals were presented in our center late (CD4+ T cell count;< 350/cu. mm) for diagnosis and care. Majority of co- infected 4 (80%) were late presented with advanced AIDS stage (CD4+ count; <200/cu.mm). Conclusions: The study showed a high percentage of HIV sero-positive & co- infected individuals. Baseline CD4+ T cell count of majority of HIV infected individuals was found to be low. Hence, more sustained and vigorous awareness campaigns & counseling still need to be done in order to promote early diagnosis and management.

Keywords: HIV/AIDS, HBsAg, co-infection, CD4+

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616 A Correlations Study on Nursing Staff's Shifts Systems, Workplace Fatigue, and Quality of Working Life

Authors: Jui Chen Wu, Ming Yi Hsu

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Background and Purpose: Shift work of nursing staff is inevitable in hospital to provide continuing medical care. However, shift work is considered as a health hazard that may cause physical and psychological problems. Serious workplace fatigue of nursing shift work might impact on family, social and work life, moreover, causes serious reduction of quality of medical care, or even malpractice. This study aims to explore relationships among nursing staff’s shift, workplace fatigue and quality of working life. Method: Structured questionnaires were used in this study to explore relationships among shift work, workplace fatigue and quality of working life in nursing staffs. We recruited 590 nursing staffs in different Community Teaching hospitals in Taiwan. Data analysed by descriptive statistics, single sample t-test, single factor analysis, Pearson correlation coefficient and hierarchical regression, etc. Results: The overall workplace fatigue score is 50.59 points. In further analysis, the score of personal burnout, work-related burnout, over-commitment and client-related burnout are 57.86, 53.83, 45.95 and 44.71. The basic attributes of nursing staff are significantly different from those of workplace fatigue with different ages, licenses, sleeping quality, self-conscious health status, number of care patients of chronic diseases and number of care people in the obstetric ward. The shift variables revealed no significant influence on workplace fatigue during the hierarchical regression analysis. About the analysis on nursing staff’s basic attributes and shift on the quality of working life, descriptive results show that the overall quality of working life of nursing staff is 3.23 points. Comparing the average score of the six aspects, the ranked average score are 3.47 (SD= .43) in interrelationship, 3.40 (SD= .46) in self-actualisation, 3.30 (SD= .40) in self-efficacy, 3.15 (SD= .38) in vocational concept, 3.07 (SD= .37) in work aspects, and 3.02 (SD= .56) in organization aspects. The basic attributes of nursing staff are significantly different from quality of working life in different marriage situations, education level, years of nursing work, occupation area, sleep quality, self-conscious health status and number of care in medical ward. There are significant differences between shift mode and shift rate with the quality of working life. The results of the hierarchical regression analysis reveal that one of the shifts variables 'shift mode' which does affect staff’s quality of working life. The workplace fatigue is negatively correlated with the quality of working life, and the over-commitment in the workplace fatigue is positively related to the vocational concept of the quality of working life. According to the regression analysis of nursing staff’s basic attributes, shift mode, workplace fatigue and quality of working life related shift, the results show that the workplace fatigue has a significant impact on nursing staff’s quality of working life. Conclusion: According to our study, shift work is correlated with workplace fatigue in nursing staffs. This results work as important reference for human resources management in hospitals to establishing a more positive and healthy work arrangement policy.

Keywords: nursing staff, shift, workplace fatigue, quality of working life

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615 Ibadan-Nigeria Citizenship Behavior Scale: Development and Validation

Authors: Benjamin O. Ehigie, Aderemi Alarape, Nyitor Shenge, Sylvester A. Okhakhume, Timileyin Fashola, Fiyinfunjah Dosumu

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Organisational citizenship behaviour (OCB) is a construct in industrial and organisational behaviour that explains a person's voluntary commitment within an organisation, which is outside the scope of his or her contractual tasks. To attain organisational effectiveness the human factor of production is inevitable, hence the importance of employee behaviour. While the concept of organisational citizenship behavior is mostly discussed in the context of the workplace, it is reasoned that the idea could be reflective in relation to national commitment. Many developing countries in Africa, including Nigeria, suffer economic hardship today not necessarily due to poor resources but bad management of the resources. The mangers of their economies are not committed to the tenets of economic growth but engrossed in fraud, corruption, bribery, and other economic vices. It is this backdrop that necessitated the development and validation of the Ibadan-Nigeria Citizenship Behaviour (I-NCB) Scale. The study adopted a cross-sectional survey (online) research design, using 2404 postgraduate students in the Premier University of the country, with 99.2% being Nigerians and 0.8% non-Nigerians. Gender composition was 1,439 (60%) males and 965 (40%) females, 1201 (50%) were employed while 1203 50% unemployed, 74.2% of the employed were in public paid employment, 19.5% in private sector, and 6.3% were self-employed. Through literature review, 78 items were generated. Using 10 lecturers and 21 students, content and face validity were established respectively. Data collected were subjected to reliability and factor analytic statistics at p < .05 level of significance. Results of the content and face validity at 80% level of item acceptance resulted to 60 items; this was further reduced to 50 after item-total correlation using r=.30 criterion. Divergent validity of r= -.28 and convergent validity of r= .44 were obtained by correlating the I-NCB scale with standardized Counterproductive work behaviour (CWB) scale and Organisational Citizenship Behaviour (OCB) scale among the workers. The reliability coefficients obtained were; Cronbach alpha of internal consistency (α = 0.941) and split-half reliability of r = 0.728. Factor analyses of the I-NCB scale with principal component and varimax rotation yielded five factors when Eigenvalue above 1 were extracted. The factors which accounted for larger proportions of the total variance were given factor names as; Altruistic, Attachment, Affective, Civic responsibility and Allegiance. As much as there are vast journals on citizenship behaviour in organisations, there exists no standardized tool to measure citizenship behaviour of a country. The Ibadan-Nigeria Citizenship Behaviour (I-NCB) scale was consequently developed. The scale could be used to select personnel into political positions and senior administrative positions among career workers in Nigeria, with the aim of determining national commitment to service.

Keywords: counterproductive work behaviour, CWB, Nigeria Citizenship Behaviour, organisational citizenship behaviour, OCB, Ibadan

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614 Implication of Woman’s Status on Child Health in India

Authors: Rakesh Mishra

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India’s Demography has always amazed the world because of its unprecedented outcomes in the presence of multifaceted socioeconomic and geographical characteristics. Being the first one to implement family panning in 1952, it occupies 2nd largest population of the world, with some of its state like Uttar Pradesh contributing 5th largest population to the world population surpassing Brazil. Being the one with higher in number it is more prone to the demographic disparity persisting into its territories brought upon by the inequalities in availability, accessibility and attainability of socioeconomic and various other resources. Fifth goal of Millennium Development Goal emphasis to improve maternal and child health across the world as Children’s development is very important for the overall development of society and the best way to develop national human resources is to take care of children. The target is to reduce the infant deaths by three quarters between 1990 and 2015. Child health status depends on the care and delivery by trained personnel, particularly through institutional facilities which is further associated with the status of the mother. However, delivery in institutional facilities and delivery by skilled personnel are rising slowly in India. The main objective of the present study is to measure the child health status on based on the educational and occupational background of the women in India. Study indicates that women education plays a very crucial role in deciding the health of the new born care and access to family planning, but the women autonomy indicates to have mixed results in different states of India. It is observed that rural women are 1.61 times more likely to exclusive breastfed their children compared to urban women. With respect to Hindu category, women belonging to other religious community were 21 percent less likely to exclusive breastfed their child. Taking scheduled caste as reference category, the odds of exclusive breastfeeding is found to be decreasing in comparison to other castes, and it is found to be significant among general category. Women of high education status have higher odds of using family planning methods in most of the southern states of India. By and large, girls and boys are about equally undernourished. Under nutrition is generally lower for first births than for subsequent births and consistently increases with increasing birth order for all measures of nutritional status. It is to be noted that at age 12-23 months, when many children are being weaned from breast milk, 30 percent of children are severely stunted and around 21 percent are severely underweight. So, this paper presents the evidence on the patterns of prevailing child health status in India and its states with reference to the mother socioeconomics and biological characteristics and examines trends in these, and discusses plausible explanations.

Keywords: immunization, exclusive breastfeeding, under five mortality, binary logistic regression, ordinal regression and life table

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613 Application of Alumina-Aerogel in Post-Combustion CO₂ Capture: Optimization by Response Surface Methodology

Authors: S. Toufigh Bararpour, Davood Karami, Nader Mahinpey

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Dependence of global economics on fossil fuels has led to a large growth in the emission of greenhouse gases (GHGs). Among the various GHGs, carbon dioxide is the main contributor to the greenhouse effect due to its huge emission amount. To mitigate the threatening effect of CO₂, carbon capture and sequestration (CCS) technologies have been studied widely in recent years. For the combustion processes, three main CO₂ capture techniques have been proposed such as post-combustion, pre-combustion and oxyfuel combustion. Post-combustion is the most commonly used CO₂ capture process as it can be readily retrofit into the existing power plants. Multiple advantages have been reported for the post-combustion by solid sorbents such as high CO₂ selectivity, high adsorption capacity, and low required regeneration energy. Chemical adsorption of CO₂ over alkali-metal-based solid sorbents such as K₂CO₃ is a promising method for the selective capture of diluted CO₂ from the huge amount of nitrogen existing in the flue gas. To improve the CO₂ capture performance, K₂CO₃ is supported by a stable and porous material. Al₂O₃ has been employed commonly as the support and enhanced the cyclic CO₂ capture efficiency of K₂CO₃. Different phases of alumina can be obtained by setting the calcination temperature of boehmite at 300, 600 (γ-alumina), 950 (δ-alumina) and 1200 °C (α-alumina). By increasing the calcination temperature, the regeneration capacity of alumina increases, while the surface area reduces. However, sorbents with lower surface areas have lower CO₂ capture capacity as well (except for the sorbents prepared by hydrophilic support materials). To resolve this issue, a highly efficient alumina-aerogel support was synthesized with a BET surface area of over 2000 m²/g and then calcined at a high temperature. The synthesized alumina-aerogel was impregnated on K₂CO₃ based on 50 wt% support/K₂CO₃, which resulted in the preparation of a sorbent with remarkable CO₂ capture performance. The effect of synthesis conditions such as types of alcohols, solvent-to-co-solvent ratios, and aging times was investigated on the performance of the support. The best support was synthesized using methanol as the solvent, after five days of aging time, and at a solvent-to-co-solvent (methanol-to-toluene) ratio (v/v) of 1/5. Response surface methodology was used to investigate the effect of operating parameters such as carbonation temperature and H₂O-to-CO₂ flowrate ratio on the CO₂ capture capacity. The maximum CO₂ capture capacity, at the optimum amounts of operating parameters, was 7.2 mmol CO₂ per gram K₂CO₃. Cyclic behavior of the sorbent was examined over 20 carbonation and regenerations cycles. The alumina-aerogel-supported K₂CO₃ showed a great performance compared to unsupported K₂CO₃ and γ-alumina-supported K₂CO₃. Fundamental performance analyses and long-term thermal and chemical stability test will be performed on the sorbent in the future. The applicability of the sorbent for a bench-scale process will be evaluated, and a corresponding process model will be established. The fundamental material knowledge and respective process development will be delivered to industrial partners for the design of a pilot-scale testing unit, thereby facilitating the industrial application of alumina-aerogel.

Keywords: alumina-aerogel, CO₂ capture, K₂CO₃, optimization

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612 The Effects of Heavy Metal and Aromatic Hydrocarbon Pollution on Bees

Authors: Katarzyna Zięba, Hajnalka Szentgyörgyi, Paweł Miśkowiec, Agnieszka Moos-Matysik

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Bees are effective pollinators of plants using by humans. However, there is a concern about the fate different species due to their recently decline. Pollution of the environment is described in the literature as one of the causes of this phenomenon. Due to human activities, heavy metals and aromatic hydrocarbons can occur in bee organisms in high concentrations. The presented study aims to provide information on how pollution affects bee quality, taking into account, also the biological differences between various groups of bees. Understanding the consequences of environmental pollution on bees can help to create and promote bee friendly habitats and actions. The analyses were carried out using two contamination gradients with 5 sites on each. The first, mainly heavy metal polluted gradient is stretching approx. 30km from the Bukowno Zinc smelter near Olkusz in the Lesser Poland Voivodship, to the north. The second cuts through the agglomeration of Kraków up to the southern borders of the Ojców National Park. The gradient near Olkusz is a well-described pollution gradient contaminated mainly by zinc, lead, and cadmium. The second gradient cut through the agglomeration of Kraków and end below the Ojców National Park. On each gradient, two bee species were installed: red mason bees (Osmia bicornis) and honey bees (Apis mellifera). Red mason bee is a polylectic, solitary bee species, widely distributed in Poland. Honey bees are a highly social species of bees, with clearly defined casts and roles in the colony. Before installing the bees in the field, samples of imagos of red mason bees and samples of pollen and imagos from each honey bee colony were analysed for zinc, lead cadmium, polycyclic and monocyclic hydrocarbons levels. After collecting the bees from the field, samples of bees and pollen samples for each site were prepared for heavy metal, monocyclic hydrocarbon, and polycyclic hydrocarbon analysis. Analyses of aromatic hydrocarbons were performed with gas chromatography coupled with a headspace sampler (HP 7694E) and mass spectrometer (MS) as detector. Monocyclic compounds were injected into column with headspace sampler while polycyclic ones with manual injector (after solid-liquid extraction with hexane). The heavy metal content (zinc, lead and cadmium) was assessed with flame atomic absorption spectroscopy (FAAS AAnalyst 300 Perkin Elmer spectrometer) according to the methods for honey and bee products described in the literature. Pollution levels found in bee bodies and imago body masses in both species, and proportion of sex in case of red mason bees were correlated with pollution levels found in pollen for each site and colony or trap nest. An attempt to pinpoint the most important form of contamination regarding bee health was also be undertaken based on the achieved results.

Keywords: heavy metals, aromatic hydrocarbons, bees, pollution

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611 Hybrid Living: Emerging Out of the Crises and Divisions

Authors: Yiorgos Hadjichristou

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The paper will focus on the hybrid living typologies which are brought about due to the Global Crisis. Mixing of the generations and the groups of people, mingling the functions of living with working and socializing, merging the act of living in synergy with the urban realm and its constituent elements will be the springboard of proposing an essential sustainable housing approach and the respective urban development. The thematic will be based on methodologies developed both on the academic, educational environment including participation of students’ research and on the practical aspect of architecture including case studies executed by the author in the island of Cyprus. Both paths of the research will deal with the explorative understanding of the hybrid ways of living, testing the limits of its autonomy. The evolution of the living typologies into substantial hybrid entities, will deal with the understanding of new ways of living which include among others: re-introduction of natural phenomena, accommodation of the activity of work and services in the living realm, interchange of public and private, injections of communal events into the individual living territories. The issues and the binary questions raised by what is natural and artificial, what is private and what public, what is ephemeral and what permanent and all the in-between conditions are eloquently traced in the everyday life in the island. Additionally, given the situation of Cyprus with the eminent scar of the dividing ‘Green line’ and the waiting of the ‘ghost city’ of Famagusta to be resurrected, the conventional way of understanding the limits and the definitions of the properties is irreversibly shaken. The situation is further aggravated by the unprecedented phenomenon of the crisis on the island. All these observations set the premises of reexamining the urban development and the respective sustainable housing in a synergy where their characteristics start exchanging positions, merge into each other, contemporarily emerge and vanish, changing from permanent to ephemeral. This fluidity of conditions will attempt to render a future of the built- and unbuilt realm where the main focusing point will be redirected to the human and the social. Weather and social ritual scenographies together with ‘spontaneous urban landscapes’ of ‘momentary relationships’ will suggest a recipe for emerging urban environments and sustainable living. Thus, the paper will aim at opening a discourse on the future of the sustainable living merged in a sustainable urban development in relation to the imminent solution of the division of island, where the issue of property became the main obstacle to be overcome. At the same time, it will attempt to link this approach to the global need for a sustainable evolution of the urban and living realms.

Keywords: social ritual scenographies, spontaneous urban landscapes, substantial hybrid entities, re-introduction of natural phenomena

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610 Genome-Wide Homozygosity Analysis of the Longevous Phenotype in the Amish Population

Authors: Sandra Smieszek, Jonathan Haines

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Introduction: Numerous research efforts have focused on searching for ‘longevity genes’. However, attempting to decipher the genetic component of the longevous phenotype have resulted in limited success and the mechanisms governing longevity remain to be explained. We conducted a genome-wide homozygosity analysis (GWHA) of the founder population of the Amish community in central Ohio. While genome-wide association studies using unrelated individuals have revealed many interesting longevity associated variants, these variants are typically of small effect and cannot explain the observed patterns of heritability for this complex trait. The Amish provide a large cohort of extended kinships allowing for in depth analysis via family-based approach excellent population due to its. Heritability of longevity increases with age with significant genetic contribution being seen in individuals living beyond 60 years of age. In our present analysis we show that the heritability of longevity is estimated to be increasing with age particularly on the paternal side. Methods: The present analysis integrated both phenotypic and genotypic data and led to the discovery of a series of variants, distinct for stratified populations across ages and distinct for paternal and maternal cohorts. Specifically 5437 subjects were analyzed and a subset of 893 successfully genotyped individuals was used to assess CHIP heritability. We have conducted the homozygosity analysis to examine if homozygosity is associated with increased risk of living beyond 90. We analyzed AMISH cohort genotyped for 614,957 SNPs. Results: We delineated 10 significant regions of homozygosity (ROH) specific for the age group of interest (>90). Of particular interest was ROH on chromosome 13, P < 0.0001. The lead SNPs rs7318486 and rs9645914 point to COL4A2 and our lead SNP. COL25A1 encodes one of the six subunits of type IV collagen, the C-terminal portion of the protein, known as canstatin, is an inhibitor of angiogenesis and tumor growth. COL4A2 mutations have been reported with a broader spectrum of cerebrovascular, renal, ophthalmological, cardiac, and muscular abnormalities. The second region of interest points to IRS2. Furthermore we built a classifier using the obtained SNPs from the significant ROH region with 0.945 AUC giving ability to discriminate between those living beyond to 90 years of age and beyond. Conclusion: In conclusion our results suggest that a history of longevity does indeed contribute to increasing the odds of individual longevity. Preliminary results are consistent with conjecture that heritability of longevity is substantial when we start looking at oldest fifth and smaller percentiles of survival specifically in males. We will validate all the candidate variants in independent cohorts of centenarians, to test whether they are robustly associated with human longevity. The identified regions of interest via ROH analysis could be of profound importance for the understanding of genetic underpinnings of longevity.

Keywords: regions of homozygosity, longevity, SNP, Amish

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609 Production of Bacillus Lipopeptides for Biocontrol of Postharvest Crops

Authors: Vivek Rangarajan, Kim G. Klarke

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With overpopulation threatening the world’s ability to feed itself, food production and protection has become a major issue, especially in developing countries. Almost one-third of the food produced for human consumption, around 1.3 billion tonnes, is either wasted or lost annually. Postharvest decay in particular constitutes a major cause of crop loss with about 20% of fruits and vegetables produced lost during postharvest storage, mainly due to fungal disease. Some of the major phytopathogenic fungi affecting postharvest fruit crops in South Africa include Aspergillus, Botrytis, Penicillium, Alternaria and Sclerotinia spp. To date control of fungal phytopathogens has primarily been dependent on synthetic chemical fungicides, but these chemicals pose a significant threat to the environment, mainly due to their xenobiotic properties and tendency to generate resistance in the phytopathogens. Here, an environmentally benign alternative approach to control postharvest fungal phytopathogens in perishable fruit crops has been presented, namely the application of a bio-fungicide in the form of lipopeptide molecules. Lipopeptides are biosurfactants produced by Bacillus spp. which have been established as green, nontoxic and biodegradable molecules with antimicrobial properties. However, since the Bacillus are capable of producing a large number of lipopeptide homologues with differing efficacies against distinct target organisms, the lipopeptide production conditions and strategy are critical to produce the maximum lipopeptide concentration with homologue ratios to specification for optimum bio-fungicide efficacy. Process conditions, and their impact on Bacillus lipopeptide production, were evaluated in fully instrumented laboratory scale bioreactors under well-regulated controlled and defined environments. Factors such as the oxygen availability and trace element and nitrate concentrations had profound influences on lipopeptide yield, productivity and selectivity. Lipopeptide yield and homologue selectivity were enhanced in cultures where the oxygen in the sparge gas was increased from 21 to 30 mole%. The addition of trace elements, particularly Fe2+, increased the total concentration of lipopeptides and a nitrate concentration equivalent to 8 g/L ammonium nitrate resulted in optimum lipopeptide yield and homologue selectivity. Efficacy studies of the culture supernatant containing the crude lipopeptide mixture were conducted using phytopathogens isolated from fruit in the field, identified using genetic sequencing. The supernatant exhibited antifungal activity against all the test-isolates, namely Lewia, Botrytis, Penicillium, Alternaria and Sclerotinia spp., even in this crude form. Thus the lipopeptide product efficacy has been confirmed to control the main diseases, even in the basic crude form. Future studies will be directed towards purification of the lipopeptide product and enhancement of efficacy.

Keywords: antifungal efficacy, biocontrol, lipopeptide production, perishable crops

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608 Green Extraction Technologies of Flavonoids Containing Pharmaceuticals

Authors: Lamzira Ebralidze, Aleksandre Tsertsvadze, Dali Berashvili, Aliosha Bakuridze

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Nowadays, there is an increasing demand for biologically active substances from vegetable, animal, and mineral resources. In terms of the use of natural compounds, pharmaceutical, cosmetic, and nutrition industry has big interest. The biggest drawback of conventional extraction methods is the need to use a large volume of organic extragents. The removal of the organic solvent is a multi-stage process. And their absolute removal cannot be achieved, and they still appear in the final product as impurities. A large amount of waste containing organic solvent damages not only human health but also has the harmful effects of the environment. Accordingly, researchers are focused on improving the extraction methods, which aims to minimize the use of organic solvents and energy sources, using alternate solvents and renewable raw materials. In this context, green extraction principles were formed. Green Extraction is a need of today’s environment. Green Extraction is the concept, and it totally corresponds to the challenges of the 21st century. The extraction of biologically active compounds based on green extraction principles is vital from the view of preservation and maintaining biodiversity. Novel technologies of green extraction are known, such as "cold methods" because during the extraction process, the temperature is relatively lower, and it doesn’t have a negative impact on the stability of plant compounds. Novel technologies provide great opportunities to reduce or replace the use of organic toxic solvents, the efficiency of the process, enhance excretion yield, and improve the quality of the final product. The objective of the research is the development of green technologies of flavonoids containing preparations. Methodology: At the first stage of the research, flavonoids containing preparations (Tincture Herba Leonuri, flamine, rutine) were prepared based on conventional extraction methods: maceration, bismaceration, percolation, repercolation. At the same time, the same preparations were prepared based on green technologies, microwave-assisted, UV extraction methods. Product quality characteristics were evaluated by pharmacopeia methods. At the next stage of the research technological - economic characteristics and cost efficiency of products prepared based on conventional and novel technologies were determined. For the extraction of flavonoids, water is used as extragent. Surface-active substances are used as co-solvent in order to reduce surface tension, which significantly increases the solubility of polyphenols in water. Different concentrations of water-glycerol mixture, cyclodextrin, ionic solvent were used for the extraction process. In vitro antioxidant activity will be studied by the spectrophotometric method, using DPPH (2,2-diphenyl-1- picrylhydrazyl) as an antioxidant assay. The advantage of green extraction methods is also the possibility of obtaining higher yield in case of low temperature, limitation extraction process of undesirable compounds. That is especially important for the extraction of thermosensitive compounds and maintaining their stability.

Keywords: extraction, green technologies, natural resources, flavonoids

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607 Antimicrobial Resistance of Acinetobacter baumannii in Veterinary Settings: A One Health Perspective from Punjab, Pakistan

Authors: Minhas Alam, Muhammad Hidayat Rasool, Mohsin Khurshid, Bilal Aslam

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The genus Acinetobacter has emerged as a significant concern in hospital-acquired infections, particularly due to the versatility of Acinetobacter baumannii in causing nosocomial infections. The organism's remarkable metabolic adaptability allows it to thrive in various environments, including the environment, animals, and humans. However, the extent of antimicrobial resistance in Acinetobacter species from veterinary settings, especially in developing countries like Pakistan, remains unclear. This study aimed to isolate and characterize Acinetobacter spp. from veterinary settings in Punjab, Pakistan. A total of 2,230 specimens were collected, including 1,960 samples from veterinary settings (nasal and rectal swabs from dairy and beef cattle), 200 from the environment, and 70 from human clinical settings. Isolates were identified using routine microbiological procedures and confirmed by polymerase chain reaction (PCR). Antimicrobial susceptibility was determined by the disc diffusion method, and minimum inhibitory concentration (MIC) was measured by the micro broth dilution method. Molecular techniques, such as PCR and DNA sequencing, were used to screen for antimicrobial-resistant determinants. Genetic diversity was assessed using standard techniques. The results showed that the overall prevalence of A. baumannii in cattle was 6.63% (65/980). However, among cattle, a higher prevalence of A. baumannii was observed in dairy cattle, 7.38% (54/731), followed by beef cattle, 4.41% (11/249). Out of 65 A. baumannii isolates, the carbapenem resistance was found in 18 strains, i.e. 27.7%. The prevalence of A. baumannii in nasopharyngeal swabs was higher, i.e., 87.7% (57/65), as compared to rectal swabs, 12.3% (8/65). Class D β-lactamases genes blaOXA-23 and blaOXA-51 were present in all the CRAB from cattle. Among carbapenem-resistant isolates, 94.4% (17/18) were positive for class B β-lactamases gene blaIMP, whereas the blaNDM-1 gene was detected in only one isolate of A. baumannii. Among 70 clinical isolates of A. baumannii, 58/70 (82.9%) were positive for the blaOXA-23-like gene, and 87.1% (61/70) were CRAB isolates. Among all clinical isolates of A. baumannii, blaOXA-51-like gene was present. Hence, the co-existence of blaOXA-23 and blaOXA-51 was found in 82.85% of clinical isolates. From the environmental settings, a total of 18 A. baumannii isolates were recovered; among these, 38.88% (7/18) strains showed carbapenem resistance. All environmental isolates of A. baumannii harbored class D β-lactamases genes, i.e., blaOXA-51 and blaOXA-23 were detected in 38.9% (7/18) isolates. Hence, the co-existence of blaOXA-23 and blaOXA-51 was found in 38.88% of isolates. From environmental settings, 18 A. baumannii isolates were recovered, with 38.88% showing carbapenem resistance. All environmental isolates harbored blaOXA-51 and blaOXA-23 genes, with co-existence in 38.88% of isolates. MLST results showed ten different sequence types (ST) in clinical isolates, with ST 589 being the most common in carbapenem-resistant isolates. In veterinary isolates, ST2 was most common in CRAB isolates from cattle. Immediate control measures are needed to prevent the transmission of CRAB isolates among animals, the environment, and humans. Further studies are warranted to understand the mechanisms of antibiotic resistance spread and implement effective disease control programs.

Keywords: Acinetobacter baumannii, carbapenemases, drug resistance, MSLT

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606 Examination of Porcine Gastric Biomechanics in the Antrum Region

Authors: Sif J. Friis, Mette Poulsen, Torben Strom Hansen, Peter Herskind, Jens V. Nygaard

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Gastric biomechanics governs a large range of scientific and engineering fields, from gastric health issues to interaction mechanisms between external devices and the tissue. Determination of mechanical properties of the stomach is, thus, crucial, both for understanding gastric pathologies as well as for the development of medical concepts and device designs. Although the field of gastric biomechanics is emerging, advances within medical devices interacting with the gastric tissue could greatly benefit from an increased understanding of tissue anisotropy and heterogeneity. Thus, in this study, uniaxial tensile tests of gastric tissue were executed in order to study biomechanical properties within the same individual as well as across individuals. With biomechanical tests in the strain domain, tissue from the antrum region of six porcine stomachs was tested using eight samples from each stomach (n = 48). The samples were cut so that they followed dominant fiber orientations. Accordingly, from each stomach, four samples were longitudinally oriented, and four samples were circumferentially oriented. A step-wise stress relaxation test with five incremental steps up to 25 % strain with 200 s rest periods for each step was performed, followed by a 25 % strain ramp test with three different strain rates. Theoretical analysis of the data provided stress-strain/time curves as well as 20 material parameters (e.g., stiffness coefficients, dissipative energy densities, and relaxation time coefficients) used for statistical comparisons between samples from the same stomach as well as in between stomachs. Results showed that, for the 20 material parameters, heterogeneity across individuals, when extracting samples from the same area, was in the same order of variation as the samples within the same stomach. For samples from the same stomach, the mean deviation percentage for all 20 parameters was 21 % and 18 % for longitudinal and circumferential orientations, compared to 25 % and 19 %, respectively, for samples across individuals. This observation was also supported by a nonparametric one-way ANOVA analysis, where results showed that the 20 material parameters from each of the six stomachs came from the same distribution with a level of statistical significance of P > 0.05. Direction-dependency was also examined, and it was found that the maximum stress for longitudinal samples was significantly higher than for circumferential samples. However, there were no significant differences in the 20 material parameters, with the exception of the equilibrium stiffness coefficient (P = 0.0039) and two other stiffness coefficients found from the relaxation tests (P = 0.0065, 0.0374). Nor did the stomach tissue show any significant differences between the three strain-rates used in the ramp test. Heterogeneity within the same region has not been examined earlier, yet, the importance of the sampling area has been demonstrated in this study. All material parameters found are essential to understand the passive mechanics of the stomach and may be used for mathematical and computational modeling. Additionally, an extension of the protocol used may be relevant for compiling a comparative study between the human stomach and the pig stomach.

Keywords: antrum region, gastric biomechanics, loading-unloading, stress relaxation, uniaxial tensile testing

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605 On the Influence of Sleep Habits for Predicting Preterm Births: A Machine Learning Approach

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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604 Sustainability Communications Across Multi-Stakeholder Groups: A Critical Review of the Findings from the Hospitality and Tourism Sectors

Authors: Frederica Pettit

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Contribution: Stakeholder involvement in CSR is essential to ensuring pro-environmental attitudes and behaviours across multi-stakeholder groups. Despite increased awareness of the benefits surrounding a collaborative approach to sustainability communications, its success is limited by difficulties engaging with active online conversations with stakeholder groups. Whilst previous research defines the effectiveness of sustainability communications; this paper contributes to knowledge through the development of a theoretical framework that explores the processes to achieving pro-environmental attitudes and behaviours in stakeholder groups. The research will also consider social media as an opportunity to communicate CSR information to all stakeholder groups. Approach: A systematic review was chosen to investigate the effectiveness of the types of sustainability communications used in the hospitality and tourism industries. The systematic review was completed using Web of Science and Scopus using the search terms “sustainab* communicat*” “effective or effectiveness,” and “hospitality or tourism,” limiting the results to peer-reviewed research. 133 abstracts were initially read, with articles being excluded for irrelevance, duplicated articles, non-empirical studies, and language. A total of 45 papers were included as part of the systematic review. 5 propositions were created based on the results of the systematic review, helping to develop a theoretical framework of the processes needed for companies to encourage pro-environmental behaviours across multi-stakeholder groups. Results: The theoretical framework developed in the paper determined the processes necessary for companies to achieve pro-environmental behaviours in stakeholders. The processes to achieving pro-environmental attitudes and behaviours are stakeholder-focused, identifying the need for communications to be specific to their targeted audience. Collaborative communications that enable stakeholders to engage with CSR information and provide feedback lead to a higher awareness of CSR shared visions and pro-environmental attitudes and behaviours. These processes should also aim to improve their relationships with stakeholders through transparency of CSR, CSR strategies that match stakeholder values and ethics whilst prioritizing sustainability as part of their job role. Alternatively, companies can prioritize pro-environmental behaviours using choice editing by mainstreaming sustainability as the only option. In recent years, there has been extensive research on social media as a viable source of sustainability communications, with benefits including direct interactions with stakeholders, the ability to enforce the authenticity of CSR activities and encouragement of pro-environmental behaviours. Despite this, there are challenges to implementing CSR, including difficulties controlling stakeholder criticisms, negative stakeholder influences and comments left on social media platforms. Conclusion: A lack of engagement with CSR information is a reoccurring reason for preventing pro-environmental attitudes and behaviours across stakeholder groups. Traditional CSR strategies contribute to this due to their inability to engage with their intended audience. Hospitality and tourism companies are improving stakeholder relationships through collaborative processes which reduce single-use plastic consumption. A collaborative approach to communications can lead to stakeholder satisfaction, leading to changes in attitudes and behaviours. Different sources of communications are accessed by different stakeholder groups, identifying the need for targeted sustainability messaging, creating benefits such as direct interactions with stakeholders, the ability to enforce the authenticity of CSR activities, and encouraging engagement with sustainability information.

Keywords: hospitality, pro-environmental attitudes and behaviours, sustainability communication, social media

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603 Advancing Equitable Healthcare for Trans and Gender-Diverse Students: A Community-Based Participatory Action Project

Authors: Al Huuskonen, Clio Lake, K. M. Naude, Polina Petlitsyna, Sorsha Henning, Julia Wimmers-Klick

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This project presents the outcomes of a community-based participatory action initiative aimed at advocating for equitable healthcare and human rights for trans, two-spirit, and gender-diverse individuals, building upon the University of British Columbia (UBC) Trans Coalition's ongoing efforts. Participatory Action Research (PAR) was chosen as the research method with the goal of improving trans rights on the UBC campus, particularly regarding equitable access to healthcare. PAR involves active community contribution throughout the research process, which in this case was done by way of liaising with student resource groups and advocacy leaders. The goals of this project were as follows: a) identify gaps in gender-affirming healthcare for UBC students by consulting the community and collaborating with UBC services, b) develop an information package outlining provincial and university-based health insurance for gender-affirming care (including hormone therapy and surgeries), FAQs, and resources for UBC's trans students, c) make this package available to UBC students and other national transgender advocacy organizations. The initiative successfully expanded the UBC AMS Student Health and Dental Plan to include gender-affirming procedural coverage, developed a care access guide for students, and advocated for improved health records inclusivity, mechanisms for trans students to report negative care experiences, and increased access to gender-affirming primary care through the on-campus health clinic. Collaboration with other universities' pride organizations and Trans Care BC yielded positive outcomes through broader coalition building and resource sharing. Ongoing efforts are underway to update provincial policies, particularly through expanding coverage under fair pharma care and addressing the compounding effects of the primary care crisis for trans individuals. The project's tangible results include improved trans rights on campus, especially in terms of healthcare access. Expanding healthcare coverage through student care benefits thousands of students, making the ability to undergo important affirming procedures more affordable. Providing students with information on extended coverage options and communication with their doctors further removes barriers to care and positively impacts student wellbeing. This initiative demonstrates the effectiveness of community-based participatory action in advancing equitable healthcare for trans and gender-diverse individuals and serves as a model for other institutions and organizations striving to promote inclusivity and advocate for marginalized populations' rights.

Keywords: equitable healthcare, trans and gender-diverse individuals, inclusivity, participatory action research project

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602 Changing the Landscape of Fungal Genomics: New Trends

Authors: Igor V. Grigoriev

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Understanding of biological processes encoded in fungi is instrumental in addressing future food, feed, and energy demands of the growing human population. Genomics is a powerful and quickly evolving tool to understand these processes. The Fungal Genomics Program of the US Department of Energy Joint Genome Institute (JGI) partners with researchers around the world to explore fungi in several large scale genomics projects, changing the fungal genomics landscape. The key trends of these changes include: (i) rapidly increasing scale of sequencing and analysis, (ii) developing approaches to go beyond culturable fungi and explore fungal ‘dark matter,’ or unculturables, and (iii) functional genomics and multi-omics data integration. Power of comparative genomics has been recently demonstrated in several JGI projects targeting mycorrhizae, plant pathogens, wood decay fungi, and sugar fermenting yeasts. The largest JGI project ‘1000 Fungal Genomes’ aims at exploring the diversity across the Fungal Tree of Life in order to better understand fungal evolution and to build a catalogue of genes, enzymes, and pathways for biotechnological applications. At this point, at least 65% of over 700 known families have one or more reference genomes sequenced, enabling metagenomics studies of microbial communities and their interactions with plants. For many of the remaining families no representative species are available from culture collections. To sequence genomes of unculturable fungi two approaches have been developed: (a) sequencing DNA from fruiting bodies of ‘macro’ and (b) single cell genomics using fungal spores. The latter has been tested using zoospores from the early diverging fungi and resulted in several near-complete genomes from underexplored branches of the Fungal Tree, including the first genomes of Zoopagomycotina. Genome sequence serves as a reference for transcriptomics studies, the first step towards functional genomics. In the JGI fungal mini-ENCODE project transcriptomes of the model fungus Neurospora crassa grown on a spectrum of carbon sources have been collected to build regulatory gene networks. Epigenomics is another tool to understand gene regulation and recently introduced single molecule sequencing platforms not only provide better genome assemblies but can also detect DNA modifications. For example, 6mC methylome was surveyed across many diverse fungi and the highest among Eukaryota levels of 6mC methylation has been reported. Finally, data production at such scale requires data integration to enable efficient data analysis. Over 700 fungal genomes and other -omes have been integrated in JGI MycoCosm portal and equipped with comparative genomics tools to enable researchers addressing a broad spectrum of biological questions and applications for bioenergy and biotechnology.

Keywords: fungal genomics, single cell genomics, DNA methylation, comparative genomics

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601 Gender Gap in Returns to Social Entrepreneurship

Authors: Saul Estrin, Ute Stephan, Suncica Vujic

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Background and research question: Gender differences in pay are present at all organisational levels, including at the very top. One possible way for women to circumvent organizational norms and discrimination is to engage in entrepreneurship because, as CEOs of their own organizations, entrepreneurs largely determine their own pay. While commercial entrepreneurship plays an important role in job creation and economic growth, social entrepreneurship has come to prominence because of its promise of addressing societal challenges such as poverty, social exclusion, or environmental degradation through market-based rather than state-sponsored activities. This opens the research question whether social entrepreneurship might be a form of entrepreneurship in which the pay of men and women is the same, or at least more similar; that is to say there is little or no gender pay gap. If the gender gap in pay persists also at the top of social enterprises, what are the factors, which might explain these differences? Methodology: The Oaxaca-Blinder Decomposition (OBD) is the standard approach of decomposing the gender pay gap based on the linear regression model. The OBD divides the gender pay gap into the ‘explained’ part due to differences in labour market characteristics (education, work experience, tenure, etc.), and the ‘unexplained’ part due to differences in the returns to those characteristics. The latter part is often interpreted as ‘discrimination’. There are two issues with this approach. (i) In many countries there is a notable convergence in labour market characteristics across genders; hence the OBD method is no longer revealing, since the largest portion of the gap remains ‘unexplained’. (ii) Adding covariates to a base model sequentially either to test a particular coefficient’s ‘robustness’ or to account for the ‘effects’ on this coefficient of adding covariates might be problematic, due to sequence-sensitivity when added covariates are correlated. Gelbach’s decomposition (GD) addresses latter by using the omitted variables bias formula, which constructs a conditional decomposition thus accounting for sequence-sensitivity when added covariates are correlated. We use GD to decompose the differences in gaps of pay (annual and hourly salary), size of the organisation (revenues), effort (weekly hours of work), and sources of finances (fees and sales, grants and donations, microfinance and loans, and investors’ capital) between men and women leading social enterprises. Database: Our empirical work is made possible by our collection of a unique dataset using respondent driven sampling (RDS) methods to address the problem that there is as yet no information on the underlying population of social entrepreneurs. The countries that we focus on are the United Kingdom, Spain, Romania and Hungary. Findings and recommendations: We confirm the existence of a gender pay gap between men and women leading social enterprises. This gap can be explained by differences in the accumulation of human capital, psychological and social factors, as well as cross-country differences. The results of this study contribute to a more rounded perspective, highlighting that although social entrepreneurship may be a highly satisfying occupation, it also perpetuates gender pay inequalities.

Keywords: Gelbach’s decomposition, gender gap, returns to social entrepreneurship, values and preferences

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600 Switchable Lipids: From a Molecular Switch to a pH-Sensitive System for the Drug and Gene Delivery

Authors: Jeanne Leblond, Warren Viricel, Amira Mbarek

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Although several products have reached the market, gene therapeutics are still in their first stages and require optimization. It is possible to improve their lacking efficiency by the use of carefully engineered vectors, able to carry the genetic material through each of the biological barriers they need to cross. In particular, getting inside the cell is a major challenge, because these hydrophilic nucleic acids have to cross the lipid-rich plasmatic and/or endosomal membrane, before being degraded into lysosomes. It takes less than one hour for newly endocytosed liposomes to reach highly acidic lysosomes, meaning that the degradation of the carried gene occurs rapidly, thus limiting the transfection efficiency. We propose to use a new pH-sensitive lipid able to change its conformation upon protonation at endosomal pH values, leading to the disruption of the lipidic bilayer and thus to the fast release of the nucleic acids into the cytosol. It is expected that this new pH-sensitive mechanism promote endosomal escape of the gene, thereby its transfection efficiency. The main challenge of this work was to design a preparation presenting fast-responding lipidic bilayer destabilization properties at endosomal pH 5 while remaining stable at blood pH value and during storage. A series of pH-sensitive lipids able to perform a conformational switch upon acidification were designed and synthesized. Liposomes containing these switchable lipids, as well as co-lipids were prepared and characterized. The liposomes were stable at 4°C and pH 7.4 for several months. Incubation with siRNA led to the full entrapment of nucleic acids as soon as the positive/negative charge ratio was superior to 2. The best liposomal formulation demonstrated a silencing efficiency up to 10% on HeLa cells, very similar to a commercial agent, with a lowest toxicity than the commercial agent. Using flow cytometry and microscopy assays, we demonstrated that drop of pH was required for the transfection efficiency, since bafilomycin blocked the transfection efficiency. Additional evidence was brought by the synthesis of a negative control lipid, which was unable to switch its conformation, and consequently exhibited no transfection ability. Mechanistic studies revealed that the uptake was mediated through endocytosis, by clathrin and caveolae pathways, as reported for previous lipid nanoparticle systems. This potent system was used for the treatment of hypercholesterolemia. The switchable lipids were able to knockdown PCSK9 expression on human hepatocytes (Huh-7). Its efficiency is currently evaluated on in vivo mice model of PCSK9 KO mice. In summary, we designed and optimized a new cationic pH-sensitive lipid for gene delivery. Its transfection efficiency is similar to the best available commercial agent, without the usually associated toxicity. The promising results lead to its use for the treatment of hypercholesterolemia on a mice model. Anticancer applications and pulmonary chronic disease are also currently investigated.

Keywords: liposomes, siRNA, pH-sensitive, molecular switch

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599 Microfungi on Sandy Beaches: Potential Threats for People Enjoying Lakeside Recreation

Authors: Tomasz Balabanski, Anna Biedunkiewicz

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Research on basic bacteriological and physicochemical parameters conducted by state institutions (Provincial Sanitary and Epidemiological Station and District Sanitary and Epidemiological Station) are limited to bathing waters under constant sanitary and epidemiological supervision. Unfortunately, no routine or monitoring tests are carried out for the presence of microfungi. This also applies to beach sand used for recreational purposes. The purpose of the planned own research was to determine the diversity of the mycobiota present on supervised and unsupervised sandy beaches, on the shores of lakes, of municipal baths used for recreation. The research material consisted of microfungi isolated from April to October 2019 from sandy beaches of supervised and unsupervised lakes located within the administrative boundaries of the city of Olsztyn (North-Eastern Poland, Europe). Four lakes, out of the fifteen available (Tyrsko, Kortowskie, Skanda, and Ukiel), whose bathing waters are subjected to routine bacteriological tests, were selected for testing. To compare the diversity of the mycobiota composition on the surface and below the sand mixing layer, samples were taken from two depths (10 cm and 50 cm), using a soil auger. Micro-fungi from sand samples were obtained by surface inoculation on an RBC medium from the 1st dilution (1:10). After incubation at 25°C for 96-144 h, the average number of CFU/dm³ was counted. Morphologically differing yeast colonies were passaged into Sabouraud agar slants with gentamicin and incubated again. For detailed laboratory analyses, culture methods (macro- and micro-cultures) and identification methods recommended in diagnostic mycological laboratories were used. The conducted research allowed obtaining 140 yeast isolates. The total average population ranged from 1.37 × 10⁻² CFU/dm³ before the bathing season (April 2019), 1.64 × 10⁻³ CFU/dm³ in the season (May-September 2019), and 1.60 × 10⁻² CFU/dm³ after the end of the season (October 2019). More microfungi were obtained from the surface layer of sand (100 isolates) than from the deeper layer (40 isolates). Reported microfungi may circulate seasonally between individual elements of the lake ecosystem. From the sand/soil from the catchment area beaches, they can get into bathing waters, stopping periodically on the coastal phyllosphere. The sand of the beaches and the phyllosphere are a kind of filter for the water reservoir. The presence of microfungi with various pathogenicity potential in these places is of major epidemiological importance. Therefore, full monitoring of not only recreational waters but also sandy beaches should be treated as an element of constant control by appropriate supervisory institutions, allowing recreational areas for public use so that the use of these places does not involve the risk of infection. Acknowledgment: 'Development Program of the University of Warmia and Mazury in Olsztyn', POWR.03.05.00-00-Z310/17, co-financed by the European Union under the European Social Fund from the Operational Program Knowledge Education Development. Tomasz Bałabański is a recipient of a scholarship from the Programme Interdisciplinary Doctoral Studies in Biology and Biotechnology (POWR.03.05.00-00-Z310/17), which is funded by the 'European Social Fund'.

Keywords: beach, microfungi, sand, yeasts

Procedia PDF Downloads 95