Search results for: effective pair potential

Authors: Jae Won Lee, Kyung Hyun Min, Hong Jae Lee, Sang Cheon Lee

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To date, imaging techniques have attracted much attention in medicine because the detection of diseases at an early stage provides greater opportunities for successful treatment. Consequently, over the past few decades, diverse imaging modalities including magnetic resonance (MR), positron emission tomography, computed tomography, and ultrasound (US) have been developed and applied widely in the field of clinical diagnosis. However, each of the above-mentioned imaging modalities possesses unique strengths and intrinsic weaknesses, which limit their abilities to provide accurate information. Therefore, multimodal imaging systems may be a solution that can provide improved diagnostic performance. Among the current medical imaging modalities, US is a widely available real-time imaging modality. It has many advantages including safety, low cost and easy access for patients. However, its low spatial resolution precludes accurate discrimination of diseased region such as cancer sites. In contrast, MR has no tissue-penetrating limit and can provide images possessing exquisite soft tissue contrast and high spatial resolution. However, it cannot offer real-time images and needs a comparatively long imaging time. The characteristics of these imaging modalities may be considered complementary, and the modalities have been frequently combined for the clinical diagnostic process. Biominerals such as calcium carbonate (CaCO3) and calcium phosphate (CaP) exhibit pH-dependent dissolution behavior. They demonstrate pH-controlled drug release due to the dissolution of minerals in acidic pH conditions. In particular, the application of this mineralization technique to a US contrast agent has been reported recently. The CaCO3 mineral reacts with acids and decomposes to generate calcium dioxide (CO2) gas in an acidic environment. These gas-generating mineralized nanoparticles generated CO2 bubbles in the acidic environment of the tumor, thereby allowing for strong echogenic US imaging of tumor tissues. On the basis of this previous work, it was hypothesized that the loading of MR contrast agents into the CaCO3 mineralized nanoparticles may be a novel strategy in designing a contrast agent for dual imaging. Herein, CaCO3 mineralized nanoparticles that were capable of generating CO2 bubbles to trigger the release of entrapped MR contrast agents in response to tumoral acidic pH were developed for the purposes of US and MR dual-modality imaging of tumors. Gd2O3 nanoparticles were selected as an MR contrast agent. A key strategy employed in this study was to prepare Gd2O3 nanoparticle-loaded mineralized nanoparticles (Gd2O3-MNPs) using block copolymer-templated CaCO3 mineralization in the presence of calcium cations (Ca2+), carbonate anions (CO32-) and positively charged Gd2O3 nanoparticles. The CaCO3 core was considered suitable because it may effectively shield Gd2O3 nanoparticles from water molecules in the blood (pH 7.4) before decomposing to generate CO2 gas, triggering the release of Gd2O3 nanoparticles in tumor tissues (pH 6.4~7.4). The kinetics of CaCO3 dissolution and CO2 generation from the Gd2O3-MNPs were examined as a function of pH and pH-dependent in vitro magnetic relaxation; additionally, the echogenic properties were estimated to demonstrate the potential of the particles for the tumor-specific US and MR imaging.

Keywords: calcium carbonate, mineralization, ultrasound imaging, magnetic resonance imaging

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

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

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

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Authors: Annis Lai Chu Fung

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While aggression had high stability throughout developmental stages and across generations, it should be the top priority of researchers and frontline helping professionals to develop prevention and intervention programme for aggressive children and children at risk of developing aggressive behaviours. Although there is a substantial amount of anti-bullying programmes, they gave disappointingly small effect sizes. The neglectful practical significance could be attributed to the overly simplistic categorisation of individuals involved as bullies or victims. In the past three decades, the distinction between reactive and proactive aggression has been well-proved. As children displaying reactively aggressive behaviours have distinct social-information processing pattern with those showing proactively aggressive behaviours, it is critical to identify the unique needs of the two subtypes accordingly when designing an intervention. The onset of reactive aggression and proactive aggression was observed at earliest in 4.4 and 6.8 years old respectively. Such findings called for a differential early intervention targeting these high-risk children. However, to the best of the author’s knowledge, the author was the first to establish an evidence-based intervention programme against reactive and proactive aggression. With the largest samples in the world, the author, in the past 10 years, explored three different approaches and their effectiveness against aggression quantitatively and qualitatively with longitudinal design. The three approaches presented are (a) cognitive-behavioral approach, (b) moral education, with Chinese marital arts and ethics as the medium, and (c) bioneurological measures (omega-3 supplementation). The studies adopted a multi-informant approach with repeated measures before and after the intervention, and follow-up assessment. Participants were recruited from primary and secondary schools in Hong Kong. In the cognitive-behavioral approach, 66 reactive aggressors and 63 proactive aggressors, aged from 11 to 17, were identified from 10,096 secondary-school children with questionnaire and subsequent structured interview. Participants underwent 10 group sessions specifically designed for each subtype of aggressor. Results revealed significant declines in aggression levels from the baseline to the follow-up assessment after 1 year. In moral education through the Chinese martial arts, 315 high-risk aggressive children, aged 6 to 12 years, were selected from 3,511 primary-school children and randomly assigned into four types of 10-session intervention group, namely martial-skills-only, martial-ethics-only, both martial-skills-and-ethics, and physical fitness (placebo). Results showed only the martial-skills-and-ethics group had a significant reduction in aggression after treatment and 6 months after treatment comparing with the placebo group. In the bioneurological approach, 218 children, aged from 8 to 17, were randomly assigned to the omega-3 supplement group and the placebo group. Results revealed that compared with the placebo group, the omega-3 supplement group had significant declines in aggression levels at the 6-month follow-up assessment. All three approaches were effective in reducing proactive and reactive aggression. Traditionally, intervention programmes against aggressive behaviour often adapted the cognitive and/or behavioural approach. However, cognitive-behavioural approach for children was recently challenged by its demanding requirement of cognitive ability. Traditional cognitive interventions may not be as beneficial to an older population as in young children. The present study offered an insightful perspective in aggression reduction measures.

Keywords: intervention, outcome effectiveness, proactive aggression, reactive aggression

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Authors: F. Portet-Koltalo, Y. Tian, I. Berger, C. Boulanger-Lecomte, A. Benamar, N. Machour

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Sediments are complex environments which can accumulate a great variety of persistent toxic contaminants such as polychlorobiphenyles (PCBs), polycyclic aromatic hydrocarbons (PAHs) and some of their more toxic degradation metabolites such as hydroxylated PAHs (OH-PAHs). Owing to their composition, fine clayey sediments can be more difficult to extract than soils using conventional solvent extraction processes. So this study aimed to compare the potential of MSPD (matrix solid phase dispersive extraction) to extract PCBs, PAHs and OH-PAHs, in comparison with microwave assisted extraction (MAE). Methodologies: MAE extraction with various solvent mixtures was used to extract PCBs, PAHs and OH-PAHs from sediments in two runs, followed by two GC-MS analyses. MSPD consisted in crushing the dried sediment with dispersive agents, introducing the mixture in cartridges and eluting the target compounds with an appropriate volume of selected solvents. So MSPD combined with cartridges containing MIPs (molecularly imprinted polymers) designed for OH-PAHs was used to extract the three families of target compounds in only one run, followed by parallel analyses in GC-MS for PAHs/PCBs and HPLC-FLD for OH-PAHs. Results: MAE extraction was optimized to extract from clayey sediments, in two runs, PAHs/PCBs in one hand and OH-PAHs in the other hand. Indeed, the best conditions of extractions (mixtures of extracting solvents, temperature) were different if we consider the polarity and the thermodegradability of the different families of target contaminants: PAHs/PCBs were better extracted using an acetone/toluene 50/50 mixture at 130°C whereas OH-PAHs were better extracted using an acetonitrile/toluene 90/10 mixture at 100°C. Moreover, the two consecutive GC-MS analyses contributed to double the total analysis time. A matrix solid phase dispersive (MSPD) extraction procedure was also optimized, with the first objective of increasing the extraction recovery yields of PAHs and PCBs from fine-grained sediment. The crushing time (2-10 min), the nature of the dispersing agents added for purifying and increasing the extraction yields (Florisil, octadecylsilane, 3-chloropropyle, 4-benzylchloride), the nature and the volume of eluting solvents (methylene chloride, hexane, hexane/acetone…) were studied. It appeared that in the best conditions, MSPD was a better extraction method than MAE for PAHs and PCBs, with respectively, mean increases of 8.2% and 71%. This method was also faster, easier and less expensive. But the other advantage of MSPD was that it allowed to introduce easily, just after the first elution process of PAHs/PCBs, a step permitting the selective recovery of OH-PAHs. A cartridge containing MIPs designed for phenols was coupled to the cartridge containing the dispersed sediment, and various eluting solvents, different from those used for PAHs and PCBs, were tested to selectively concentrate and extract OH-PAHs. Thereafter OH-PAHs could be analyzed at the same time than PAHs and PCBs: the OH-PAH extract could be analyzed with HPLC-FLD, whereas the PAHs/PCBs extract was analyzed with GC-MS, adding only few minutes more to the total duration of the analytical process. Conclusion: MSPD associated to MIPs appeared to be an easy, fast and low expensive method, able to extract in one run a complex mixture of toxic apolar and more polar contaminants present in clayey fine-grained sediments, an environmental matrix which is generally difficult to analyze.

Keywords: contaminated fine-grained sediments, matrix solid phase dispersive extraction, microwave assisted extraction, molecularly imprinted polymers, multi-pollutant analysis

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Authors: Md Shafiqul Bari, Md Shafiqul Islam Sikdar

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This paper summarizes the current knowledge on Agroforestry practices in the Barind tract of Bangladesh. The part of greater Rajshahi, Dinajpur, Rangpur and Bogra district of Bangladesh is geographically identified as the Barind tract. The hard red soil of these areas is very significant in comparison to that of the other parts of the country. A typical dry climate with comparatively high temperature prevails in the Barind area. Scanty rainfall and excessive extraction of groundwater have created an alarming situation among the Barind people and others about irrigation to the rice field. In addition, the situation may cause an adverse impact on the people whose livelihood largely depends on agriculture. The groundwater table has been declined by at least 10 to 15 meters in some areas of the Barind tract during the last 20 years. Due to absent of forestland in the Barind tract, the soil organic carbon content can decrease more rapidly because of the higher rate of decomposition. The Barind soils are largely carbon depleted but can be brought back to carbon-carrying capacity by bringing under suitable Agroforestry systems. Agroforestry has tremendous potential for carbon sequestration not only in above C biomass but also root C biomass in deeper soil depths. Agroforestry systems habitually conserve soil organic carbon and maintain a great natural nutrient pool. Cultivation of trees with arable crops under Agroforestry systems help in improving soil organic carbon content and sequestration carbon, particularly in the highly degraded Barind lands. Agroforestry systems are a way of securing the growth of cash crops that may constitute an alternative source of income in moments of crisis. Besides being a source of fuel wood, a greater presence of trees in cropping system contributes to decreasing temperatures and to increasing rainfall, thus contrasting the negative environmental impact of climate changes. In order to fulfill the objectives of this study, two experiments were conducted. The first experiment was survey on the impact of existing agroforestry system on the livelihood security in the Barind tract of Bangladesh and the second one was the role of agroforestry system on the improvement of soil properties in a multilayered coconut orchard. Agroforestry systems have been generated a lot of employment opportunities in the Barind area. More crops mean involvement of more people in various activities like involvements in dairying, sericulture, apiculture and additional associated agro-based interventions. Successful adoption of Agroforestry practices in the Barind area has shown that the Agroforestry practitioners of this area were very sound positioned economically, and had added social status too. However, from the findings of the present study, it may be concluded that the majority rural farmers of the Barind tract of Bangladesh had a very good knowledge and medium extension contact related to agroforestry production system. It was also observed that 85 per cent farmers followed agroforestry production system and received benefits to a higher extent. Again, from the research study on orchard based mutistoried agroforestry cropping system, it was evident that there was an important effect of agroforestry cropping systems on the improvement of soil chemical properties. As a result, the agroforestry systems may be helpful to attain the development objectives and preserve the biosphere core.

Keywords: agroforestry systems, Barind tract, carbon sequestration, climate changes

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Authors: Rebekah Granger Ellis, Richard B. Speaker, Pat Austin

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This study used two psychological scales to examine the level of social and emotional intelligence and moral judgment of over 500 gifted and talented high school students in various academic and creative arts programs in a large metropolitan area in the southeastern United States. For decades, numerous models and programs purporting to encourage socio-affective characteristics of adolescent development have been explored in curriculum theory and design. Socio-affective merges social, emotional, and moral domains. It encompasses interpersonal relations and social behaviors; development and regulation of emotions; personal and gender identity construction; empathy development; moral development, thinking, and judgment. Examining development in these socio-affective domains can provide insight into why some gifted and talented adolescents are not successful in adulthood despite advanced IQ scores. Particularly whether nonintellectual characteristics of gifted and talented individuals, such as emotional, social and moral capabilities, are as advanced as their intellectual abilities and how these are related to each other. Unique characteristics distinguish gifted and talented individuals; these may appear as strengths, but there is the potential for problems to accompany them. Although many thrive in their school environments, some gifted students struggle rather than flourish. In the socio-affective domain, these adolescents face special intrapersonal, interpersonal, and environmental problems. Gifted individuals’ cognitive, psychological, and emotional development occurs asynchronously, in multidimensional layers at different rates and unevenly across ability levels. Therefore, it is important to examine the long-term effects of participation in various gifted and talented programs on the socio-affective development of gifted and talented adolescents. This quasi-experimental longitudinal study examined students in several gifted and talented education programs (creative arts school, urban charter schools, and suburban public schools) for (1) socio-affective development level and (2) whether a particular gifted and talented program encourages developmental growth. The following research questions guided the study: (1) How do academically and artistically talented gifted 10th and 11th grade students perform on psychometric scales of social and emotional intelligence and moral judgment? Do they differ from their age or grade normative sample? Are their gender differences among gifted students? (2) Does school environment impact 10th and 11th grade gifted and talented students’ socio-affective development? Do gifted adolescents who participate in a particular school gifted program differ in their developmental profiles of social and emotional intelligence and moral judgment? Students’ performances on psychometric instruments were compared over time and by type of program. Participants took pre-, mid-, and post-tests over the course of an academic school year with Defining Issues Test (DIT-2) assessing moral judgment and BarOn EQ-I: YV assessing social and emotional intelligence. Based on these assessments, quantitative differences in growth on psychological scales (individual and school) were examined. Change scores between schools were also compared. If a school showed change, artifacts (culture, curricula, instructional methodology) provided insight as to environmental qualities that produced this difference.

Keywords: gifted and talented education, moral development, socio-affective development, socio-affective education

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Authors: Juliane Toce, Steven Holochwost

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A substantial and growing literature has demonstrated that arts education benefits young children’s socioemotional and cognitive development. Less is known about the capacity of arts-integrated instruction to yield benefits to similar domains, particularly among demographically and socioeconomically diverse groups of young children. However, the small literature on this topic suggests that arts-integrated instruction may foster young children’s socioemotional and cognitive development by presenting opportunities to 1) engage in instructional content in diverse ways, 2) experience and regulate strong emotions, 3) experience growth-oriented feedback, and 4) engage in collaborative work with peers. Start with the Art is a new program of arts-integrated instruction currently being implemented in four schools in a school district that serves students from a diverse range of backgrounds. The program employs a co-teaching model in which teaching artists and classroom teachers engage in collaborative lesson planning and instruction over the course of the academic year and is currently the focus of an impact study featuring a randomized-control design, as well as an implementation study, both of which are funded through an Educational Innovation and Research grant from the United States Department of Education. The paper will present the early results from the Start with the Art implementation study. These results will provide an overview of the extent to which the program was implemented in accordance with design, with a particular emphasis on the degree to which the four opportunities enumerated above (e.g., opportunities to engage in instructional content in diverse ways) were presented to students. There will be a review key factors that may influence the fidelity of implementation, including classroom teachers’ reception of the program and the extent to which extant conditions in the classroom (e.g., the overall level of classroom organization) may have impacted implementation fidelity. With the explicit purpose of creating a program that values and meets the needs of the teachers and students, Start with the Art incorporates the feedback from individuals participating in the intervention. Tracing its trajectory from inception to ongoing development and examining the adaptive changes made in response to teachers' transformative experiences in the post-pandemic classroom, Start with the Art continues to solicit input from experts in integrating artistic content into core curricula within educational settings catering to students from under-represented backgrounds in the arts. Leveraging the input from this rich consortium of experts has allowed for a comprehensive evaluation of the program’s implementation. The early findings derived from the implementation study emphasize the potential of arts-integrated instruction to incorporate restorative practices. Such practices serve as a crucial support system for both students and educators, providing avenues for children to express themselves, heal emotionally, and foster social development, while empowering teachers to create more empathetic, inclusive, and supportive learning environments. This all-encompassing analysis spotlights Start with the Art’s adaptability to any learning environment through the program’s effectiveness, resilience, and its capacity to transform - through art - the classroom experience within the ever-evolving landscape of education.

Keywords: arts-integration, social emotional learning, diverse learners, co-teaching, teaching artists, post-pandemic teaching

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Authors: Juan Pablo Bertuzzi, Mauro Chiarella

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Employing a communication studies perspective and a socio-technological approach, this paper introduces a theoretical framework for understanding the concept of hybrid world; the avatarization phenomena; and the communicational archetype of co-hybridization. This analysis intends to make a contribution to future design of virtual reality experimental applications. Ultimately, this paper presents an ongoing research project that proposes the study of human-avatar interactions in digital educational environments, as well as an innovative reflection on inner digital communication. The aforementioned project presents the analysis of human-avatar interactions, through the development of an interactive experience in virtual reality. The goal is to generate an innovative communicational dimension that could reinforce the hypotheses presented throughout this paper. Being thought for its initial application in educational environments, the analysis and results of this research are dependent and have been prepared in regard of a meticulous planning of: the conception of a 3D digital platform; the interactive game objects; the AI or computer avatars; the human representation as hybrid avatars; and lastly, the potential of immersion, ergonomics and control diversity that can provide the virtual reality system and the game engine that were chosen. The project is divided in two main axes: The first part is the structural one, as it is mandatory for the construction of an original prototype. The 3D model is inspired by the physical space that belongs to an academic institution. The incorporation of smart objects, avatars, game mechanics, game objects, and a dialogue system will be part of the prototype. These elements have all the objective of gamifying the educational environment. To generate a continuous participation and a large amount of interactions, the digital world will be navigable both, in a conventional device and in a virtual reality system. This decision is made, practically, to facilitate the communication between students and teachers; and strategically, because it will help to a faster population of the digital environment. The second part is concentrated to content production and further data analysis. The challenge is to offer a scenario’s diversity that compels users to interact and to question their digital embodiment. The multipath narrative content that is being applied is focused on the subjects covered in this paper. Furthermore, the experience with virtual reality devices proposes users to experiment in a mixture of a seemingly infinite digital world and a small physical area of movement. This combination will lead the narrative content and it will be crucial in order to restrict user’s interactions. The main point is to stimulate and to grow in the user the need of his hybrid avatar’s help. By building an inner communication between user’s physicality and user’s digital extension, the interactions will serve as a self-guide through the gameworld. This is the first attempt to make explicit the avatarization phenomena and to further analyze the communicational archetype of co-hybridization. The challenge of the upcoming years will be to take advantage from these forms of generalized avatarization, in order to create awareness and establish innovative forms of hybridization.

Keywords: avatar, hybrid worlds, socio-technology, virtual reality

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Authors: Aberdam Edith, Sangari Linda, Petit Isabelle, Aberdam Daniel

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Background and Rationale: Transparent avascularised cornea is essential for normal vision and depends on limbal stem cells (LSC) that reside between the cornea and the conjunctiva. Ocular burns or injuries may destroy the limbus, causing limbal stem cell deficiency (LSCD). The cornea becomes vascularised by invaded conjunctival cells, the stroma is scarring, resulting in corneal opacity and loss of vision. Grafted autologous limbus or cultivated autologous LCS can restore the vision, unless the two eyes are affected. Alternative cellular sources have been tested in the last decades, including oral mucosa or hair follicle epithelial cells. However, only partial success has been achieved by the use of these cells since they were not able to uniformly commit into corneal epithelial cells. Human pluripotent stem cells (iPSC) display both unlimited growth capacity and ability to differentiate into any cell type. Our goal was to design a standardized and reproducible protocol to produce transplantable autologous LSC from patients through cell reprogramming technology. Methodology: First, keratinocyte primary culture was established from a small number of plucked hair follicles of healthy donors. The resulting epithelial cells were reprogrammed into induced pluripotent stem cells (iPSCs) and further differentiate into corneal epithelial cells (CEC), according to a robust protocol that recapitulates the main step of corneal embryonic development. qRT-PCR analysis and immunofluorescent staining during the course of differentiation confirm the expression of stage specific markers of corneal embryonic lineage. First appear ectodermal progenitor-specific cytokeratins K8/K18, followed at day 7 by limbal-specific PAX6, TP63 and cytokeratins K5/K14. At day 15, K3/K12+-corneal cells are present. To amplify the iPSC-derived LSC (named COiPSC), intact small epithelial colonies were detached and cultivated in limbal cell-specific medium. In that culture conditions, the COiPSC can be frozen and thaw at any passage, while retaining their corneal characteristics for at least eight passages. To evaluate the potential of COiPSC as an alternative ocular toxicity model, COiPSC were treated at passage P0 to P4 with increasing amounts of SDS and Benzalkonium. Cell proliferation and apoptosis of treated cells was compared to LSC and the SV40-immortalized human corneal epithelial cell line (HCE) routinely used by cosmetological industrials. Of note, HCE are more resistant to toxicity than LSC. At P0, COiPSC were systematically more resistant to chemical toxicity than LSC and even to HCE. Remarkably, this behavior changed with passage since COiPSC at P2 became identical to LSC and thus closer to physiology than HCE. Comparative transcriptome analysis confirmed that COiPSC from P2 are similar to a mixture of LSC and CEC. Finally, by organotypic reconstitution assay, we demonstrated the ability of COiPSC to produce a 3D corneal epithelium on a stromal equivalent made of keratocytes. Conclusion: COiPSC could become valuable for two main applications: (1) an alternative robust tool to perform, in a reproducible and physiological manner, toxicity assays for cosmetic products and pharmacological tests of drugs. (2). COiPSC could become an alternative autologous source for cornea transplantation for LSCD.

Keywords: Limbal stem cell deficiency, iPSC, cornea, limbal stem cells

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Authors: Mohammadjavad Sotoudeheian

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Heart failure (HF) prevalence, as a global cardiovascular problem, is increasing gradually. A variety of molecular mechanisms contribute to HF. Proteins involved in cardiac contractility regulation, such as ion channels and calcium handling proteins, are altered. Additionally, epigenetic modifications and gene expression can lead to altered cardiac function. Moreover, inflammation and oxidative stress contribute to HF. The progression of HF can be attributed to mitochondrial dysfunction that impairs energy production and increases apoptosis. Molecular mechanisms such as these contribute to the development of cardiomyocyte defects and HF and can be therapeutically targeted. The heart's contractile function is controlled by cardiomyocytes. Calpain, and its related molecules, including Bax, VEGF, and AMPK, are among the proteins involved in regulating cardiomyocyte function. Apoptosis is facilitated by Bax. Cardiomyocyte apoptosis is regulated by this protein. Furthermore, cardiomyocyte survival, contractility, wound healing, and proliferation are all regulated by VEGF, which is produced by cardiomyocytes during inflammation and cytokine stress. Cardiomyocyte proliferation and survival are also influenced by AMPK, an enzyme that plays an active role in energy metabolism. They all play key roles in apoptosis, angiogenesis, hypertrophy, and metabolism during myocardial inflammation. The role of calpains has been linked to several molecular pathways. The calpain pathway plays an important role in signal transduction and apoptosis, as well as autophagy, endocytosis, and exocytosis. Cell death and survival are regulated by these calcium-dependent cysteine proteases that cleave proteins. As a result, protein fragments can be used for various cellular functions. By cleaving adhesion and motility proteins, calcium proteins also contribute to cell migration. HF may be brought about by calpain-mediated pathways. Many physiological processes are mediated by the calpain molecular pathways. Signal transduction, cell death, and cell migration are all regulated by these molecular pathways. Calpain is activated by calcium binding to calmodulin. In the presence of calcium, calmodulin activates calpain. Calpains are stimulated by calcium, which increases matrix metalloproteinases (MMPs). In order to develop novel treatments for these diseases, we must understand how this pathway works. A variety of myocardial remodeling processes involve calpains, including remodeling of the extracellular matrix and hypertrophy of cardiomyocytes. Calpains also play a role in maintaining cardiac homeostasis through apoptosis and autophagy. The development of HF may be in part due to calpain-mediated pathways promoting cardiomyocyte death. Numerous studies have suggested the importance of the Ca2+ -dependent protease calpain in cardiac physiology and pathology. Therefore, it is important to consider this pathway to develop and test therapeutic options in humans that targets calpain in HF. Apoptosis, autophagy, endocytosis, exocytosis, signal transduction, and disease progression all involve calpain molecular pathways. Therefore, it is conceivable that calpain inhibitors might have therapeutic potential as they have been investigated in preclinical models of several conditions in which the enzyme has been implicated that might be treated with them. Ca 2+ - dependent proteases and calpains contribute to adverse ventricular remodeling and HF in multiple experimental models. In this manuscript, we will discuss the calpain molecular pathway's important roles in HF development.

Keywords: calpain, heart failure, autophagy, apoptosis, cardiomyocyte

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Authors: Sum Lau, Wing Chung Cleo Lau, Wing Yan Chu, Long Ching Ip, Wan Yin Lo, Jo Long Sam Yau, Ka Ho Hui, Sze Yi Mak

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Biomedical engineering (BME) is an interdisciplinary subject where knowledge about biology and medicine is applied to novel applications, solving clinical problems. This subject is crucial for cities such as Hong Kong, where the burden on the medical system is rising due to reasons like the ageing population. Hong Kong, who is actively boosting technological advancements in recent years, sets BME, or biotechnology, as a major category, as reflected in the 2018-19 Budget, where biotechnology was one of the four pillars for development. Over the years, while resources in terms of money and space have been provided, there has been a lack of talents expressed by both the academia and industry. While exogenous factors, such as COVID, may have hindered talents from outside Hong Kong to come, endogenous factors should also be considered. In particular, since there are already a few local universities offering BME programmes, their curriculum or style of education requires to be reviewed to intensify the network of the BME community and support post-academic career development. It was observed that while undergraduate (UG) studies focus on knowledge teaching with some technical training and postgraduate (PG) programmes concentrate on upstream research, the programmes are generally confined to the academic sector and lack connections to the industry. In light of that, a “Biomedical Innovation and Outreach Programme 2022” (“B.I.O.2022”) was held to connect students and professors from academia with clinicians and engineers from the industry, serving as a comparative approach to conventional education methods (UG and PG programmes from tertiary institutions). Over 100 participants, including undergraduates, postgraduates, secondary school students, researchers, engineers, and clinicians, took part in various outreach events such as conference and site visits, all held from June to July 2022. As a case study, this programme aimed to tackle the aforementioned problems with the theme of “4Cs” (connection, communication, collaboration, and commercialisation). The effectiveness of the programme is investigated by its ability to serve as an adult and continuing education and the effectiveness of causing social change to tackle current societal challenges, with the focus on tackling the lack of talents engaging in biomedical engineering. In this study, B.I.O.2022 is found to be able to complement the traditional educational methods, particularly in terms of knowledge exchange between the academia and the industry. With enhanced communications between participants from different career stages, there were students who followed up to visit or even work with the professionals after the programme. Furthermore, connections between the academia and industry could foster the generation of new knowledge, which ultimately pointed to commercialisation, adding value to the BME industry while filling the gap in terms of human resources. With the continuation of events like B.I.O.2022, it provides a promising starting point for the development and relationship strengthening of a BME community in Hong Kong, and shows potential as an alternative way of adult education or learning with societal benefits.

Keywords: biomedical engineering, adult education for social change, comparative methods and principles, lifelong learning, faced problems, promises, challenges and pitfalls

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Authors: Pritha Niraj Arya

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Education plays a crucial role in societal evolution by promoting economic expansion and creativity. The varied demands of students in India’s higher education setting signify inclusive and efficient teaching methods. The present study examined how teaching styles, self-efficacy, and personality traits interact among Indian higher education faculty members and how these factors collectively affect pedagogical practices. Specifically, the research explored differences in personality traits -agreeableness, conscientiousness, neuroticism, openness, and extraversion- between teachers with high and low self-efficacy and examined how these traits shape teaching strategies, either student-focused or teacher-focused. Data collection took place for three months, ensuring confidentiality and ethical compliance. 268 faculty members from Indian higher education institutions participated in this comparative study. An online questionnaire was used to gather data in which participants completed three well-established tools: the approaches to teaching inventory, which measures teaching styles; the teacher self-efficacy questionnaire, which measures self-efficacy levels; and the big five inventory, which measures personality traits. The results showed that while teachers with low self-efficacy had higher levels of neuroticism, those with high self-efficacy scored much higher on traits such as agreeableness, conscientiousness, openness, and extraversion. Despite the traditional belief that high self-efficacy is only associated with student-focused teaching, the findings suggest that teachers with high self-efficacy have cognitive flexibility, which enables them to skillfully use both teacher-focused and student-focused approaches to cater to a wide range of classroom needs. Teachers with low self-efficacy, on the other hand, are less flexible and adopt fewer different strategies in their teaching practice. The findings challenge simplistic associations between self-efficacy and teaching strategies, emphasising that high self-efficacy promotes adaptability rather than a fixed preference for specific teaching methods. This adaptability is crucial in India’s diverse educational settings, where teachers must balance standardised curricula with the varied learning needs of students. This study highlights the importance of integrating personality traits and self-efficacy into teacher training programs. By promoting self-efficacy and tailoring professional development to consider individual personality traits, institutions can enhance teachers’ teaching flexibility, hence improving student engagement and learning outcomes. These findings have practical implications for teacher education, suggesting that adopting cognitive flexibility among teachers can improve instructional quality and classroom dynamics. To gain a deeper knowledge of how personality traits and self-efficacy impact teaching practices over time, future research should investigate causal relationships using longitudinal studies. Examining external factors like institutional policies, availability of resources, and cultural settings will help to clarify the dynamics at play. Furthermore, this study emphasises the need to strike a balance between teacher-focused and student-focused approaches to provide a comprehensive education that covers both conceptual understanding and the delivery of key information. This study offers insights into how the Indian educational system is changing and how, to achieve global standards, effective teaching techniques are becoming increasingly important. This study promotes the larger objective of educational excellence by exploring the interaction of internal and external factors impacting teaching styles and providing practical policy and practice recommendations.

Keywords: higher education, personality traits, self-efficacy, teaching styles

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Authors: Majed Al Otaibi, Melissa Lessard-Beaudoin, Amel Loudghi, Raphael Chouinard-Watkins, Melanie Plourde, Frederic Calon, C. Alexandre Castellano, Stephen Cunnane, Helene Payette, Pierrette Gaudreau, Denis Gris, Rona K. Graham

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Introduction: Alzheimer disease (AD) is a chronic disorder that affects millions of individuals worldwide. Symptoms include memory dysfunction, and also alterations in attention, planning, language and overall cognitive function. Olfactory dysfunction is a common symptom of several neurological disorders including AD. Studying the mechanisms underlying the olfactory dysfunction may therefore lead to the discovery of potential biomarkers and/or treatments for neurodegenerative diseases. Objectives: To determine if olfactory dysfunction predicts future cognitive impairment in the aging population and to characterize the olfactory system in a murine model expressing a genetic factor of AD. Method: For the human study, quantitative olfactory tests (UPSIT and OMT) have been done on 93 subjects (aged 80 to 94 years) from the Quebec Longitudinal Study on Nutrition and Successful Aging (NuAge) cohort accepting to participate in the ORCA secondary study. The telephone Modified Mini Mental State examination (t-MMSE) was used to assess cognition levels, and an olfactory self-report was also collected. In a separate cohort, olfactory cortical volume was calculated using MRI results from healthy old adults (n=25) and patients with AD (n=18) using the AAL single-subject atlas and performed with the PNEURO tool (PMOD 3.7). For the murine study, we are using Western blotting, RT-PCR and immunohistochemistry. Result: Human Study: Based on the self-report, 81% of the participants claimed to not suffer from any problem with olfaction. However, based on the UPSIT, 94% of those subjects showed a poor olfactory performance and different forms of microsmia. Moreover, the results confirm that olfactory function declines with age. We also detected a significant decrease in olfactory cortical volume in AD individuals compared to controls. Murine study: Preliminary data demonstrate there is a significant decrease in expression levels of the proform of caspase-3 and the caspase substrate STK3, in the olfactory bulb of mice expressing human APOE4 compared with controls. In addition, there is a significant decrease in the expression level of the caspase-9 proform and caspase-8 active fragment. Analysis of the mature neuron marker, NeuN, shows decreased expression levels of both isoforms. The data also suggest that Iba-1 immunostaining is increased in the olfactory bulb of APOE4 mice compared to wild type mice. Conclusions: The activation of caspase-3 may be the cause of the decreased levels of STK3 through caspase cleavage and may play role in the inflammation observed. In the clinical study, our results suggest that seniors are unaware of their olfactory function status and therefore it is not sufficient to measure olfaction using the self-report in the elderly. Studying olfactory function and cognitive performance in the aging population will help to discover biomarkers in the early stage of the AD.

Keywords: Alzheimer's disease, APOE4, cognition, caspase, brain atrophy, neurodegenerative, olfactory dysfunction

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

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

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

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Authors: Bianca Peterson, Tomasz J. Sańko, Carlos C. Bezuidenhout, Johnnie Van Den Berg

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Busseola fusca (Fuller) (Lepidoptera: Noctuidae), the maize stem borer, is a major pest in sub-Saharan Africa. It causes economic damage to maize and sorghum crops and has evolved non-recessive resistance to genetically modified (GM) maize expressing the Cry1Ab insecticidal toxin. Since B. fusca is a non-model organism, very little genomic information is publicly available, and is limited to some cytochrome c oxidase I, cytochrome b, and microsatellite data. The biology of B. fusca is well-described, but still poorly understood. This, in combination with its larval-specific behavior, may pose problems for limiting the spread of current resistant B. fusca populations or preventing resistance evolution in other susceptible populations. As part of on-going research into resistance evolution, B. fusca larvae were collected from Bt and non-Bt maize in South Africa, followed by RNA isolation (15 specimens) and sequencing on the Illumina HiSeq 2500 platform. Quality of reads was assessed with FastQC, after which Trimmomatic was used to trim adapters and remove low quality, short reads. Trinity was used for the de novo assembly, whereas TransRate was used for assembly quality assessment. Transcript identification employed BLAST (BLASTn, BLASTp, and tBLASTx comparisons), for which two libraries (nucleotide and protein) were created from 3.27 million lepidopteran sequences. Several transcripts that have previously been implicated in Cry toxin resistance was identified for B. fusca. These included aminopeptidase N, cadherin, alkaline phosphatase, ATP-binding cassette transporter proteins, and mitogen-activated protein kinase. MEGA7 was used to align these transcripts to reference sequences from Lepidoptera to detect mutations that might potentially be contributing to Cry toxin resistance in this pest. RSEM and Bioconductor were used to perform differential gene expression analysis on groups of B. fusca larvae challenged and unchallenged with the Cry1Ab toxin. Pairwise expression comparisons of transcripts that were at least 16-fold expressed at a false-discovery corrected statistical significance (p) ≤ 0.001 were extracted and visualized in a hierarchically clustered heatmap using R. A total of 329,194 transcripts with an N50 of 1,019 bp were generated from the over 167.5 million high-quality paired-end reads. Furthermore, 110 transcripts were over 10 kbp long, of which the largest one was 29,395 bp. BLAST comparisons resulted in identification of 157,099 (47.72%) transcripts, among which only 3,718 (2.37%) were identified as Cry toxin receptors from lepidopteran insects. According to transcript expression profiles, transcripts were grouped into three subclusters according to the similarity of their expression patterns. Several immune-related transcripts (pathogen recognition receptors, antimicrobial peptides, and inhibitors) were up-regulated in the larvae feeding on Bt maize, indicating an enhanced immune status in response to toxin exposure. Above all, extremely up-regulated arylphorin genes suggest that enhanced epithelial healing is one of the resistance mechanisms employed by B. fusca larvae against the Cry1Ab toxin. This study is the first to provide a resource base and some insights into a potential mechanism of Cry1Ab toxin resistance in B. fusca. Transcriptomic data generated in this study allows identification of genes that can be targeted by biotechnological improvements of GM crops.

Keywords: epithelial healing, Lepidoptera, resistance, transcriptome

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Authors: Ali R. Alshawawreh, Francisco Liébana-Cabanillas, Francisco J. Blanco-Encomienda

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Collaboration between marketing and business intelligence (BI) is crucial in today's ever-evolving business landscape. These two domains play pivotal roles in molding customers' experiential knowledge. Marketing insights offer valuable information regarding customer needs, preferences, and behaviors. Conversely, BI facilitates data-driven decision-making, leading to heightened operational efficiency, product quality, and customer satisfaction. Customer experiential knowledge (CEK) encompasses customers' implicit comprehension of consumption experiences influenced by diverse factors, including social and cultural influences. This study primarily focuses on telecommunications companies in Jordan, scrutinizing how experiential customer knowledge mediates the relationship between marketing intelligence and business intelligence. Drawing on theoretical frameworks such as the resource-based view (RBV) and service-dominant logic (SDL), the research aims to comprehend how organizations utilize their resources, particularly knowledge, to foster Evolution. Employing a quantitative research approach, the study collected and analyzed primary data to explore hypotheses. Structural equation modeling (SEM) facilitated by Smart PLS software evaluated the relationships between the constructs, followed by mediation analysis to assess the indirect associations in the model. The study findings offer insights into the intricate dynamics of organizational Creation, uncovering the interconnected relationships between business intelligence, customer experiential knowledge-based innovation (CEK-DI), marketing intelligence (MI), and product and service innovation (PSI), underscoring the pivotal role of advanced intelligence capabilities in developing innovative practices rooted in a profound understanding of customer experiences. Furthermore, the positive impact of BI on PSI reaffirms the significance of data-driven decision-making in shaping the innovation landscape. The significant impact of CEK-DI on PSI highlights the critical role of customer experiences in driving an organization. Companies that actively integrate customer insights into their opportunity creation processes are more likely to create offerings that match customer expectations, which drives higher levels of product and service sophistication. Additionally, the positive and significant impact of MI on CEK-DI underscores the critical role of market insights in shaping evolutionary strategies. While the relationship between MI and PSI is positive, the slightly weaker significance level indicates a subtle association, suggesting that while MI contributes to the development of ideas, In conclusion, the study emphasizes the fundamental role of intelligence capabilities, especially artificial intelligence, emphasizing the need for organizations to leverage market and customer intelligence to achieve effective and competitive innovation practices. Collaborative efforts between marketing and business intelligence serve as pivotal drivers of development, influencing customer experiential knowledge and shaping organizational strategies and practices. Future research could adopt longitudinal designs and gather data from various sectors to offer broader insights. Additionally, the study focuses on the effects of marketing intelligence, business intelligence, customer experiential knowledge, and innovation, but other unexamined variables may also influence innovation processes. Future studies could investigate additional factors, mediators, or moderators, including the role of emerging technologies like AI and machine learning in driving innovation.

Keywords: marketing intelligence, business intelligence, product, customer experiential knowledge-driven innovation

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Authors: Syed Jamil H. Kazmi, Faiza Sarwar

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The Indus delta of Sindh Province forms the largest creeks zone of Pakistan. The Sindh coast starts from the mouth of Hab River and terminates at Sir Creek area. In this paper, we have considered the major creeks from the site of Bin Qasim Port in Karachi to Jetty of Keti Bunder in Thatta District. A general decline in the mangrove forest has been observed that within a span of last 25 years. The unprecedented human interventions damage the creeks habitat badly which includes haphazard urban development, industrial and sewage disposal, illegal cutting of mangroves forest, reduced and inconsistent fresh water flow mainly from Jhang and Indus rivers. These activities not only harm the creeks habitat but affected the fish stock substantially. Fishing is the main livelihood of coastal people but with the above-mentioned threats, it is also under enormous pressure by fish catches resulted in unchecked overutilization of the fish resources. This pressure is almost unbearable when it joins with deleterious fishing methods, uncontrolled fleet size, increase trash and by-catch of juvenile and illegal mesh size. Along with these anthropogenic interventions study area is under the red zone of tropical cyclones and active seismicity causing floods, sea intrusion, damage mangroves forests and devastation of fish stock. In order to sustain the natural resources of the Indus Creeks, this study was initiated with the support of FAO, WWF and NIO, the main purpose was to develop a Geo-Spatial dataset for fish stock assessment. The study has been spread over a year (2013-14) on monthly basis which mainly includes detailed fish stock survey, water analysis and few other environmental analyses. Environmental analysis also includes the habitat classification of study area which has done through remote sensing techniques for 22 years’ time series (1992-2014). Furthermore, out of 252 species collected, fifteen species from estuarine and marine groups were short-listed to measure the weight, health and growth of fish species at each creek under GIS data through SPSS system. Furthermore, habitat suitability analysis has been conducted by assessing the surface topographic and aspect derivation through different GIS techniques. The output variables then overlaid in GIS system to measure the creeks productivity. Which provided the results in terms of subsequent classes: extremely productive, highly productive, productive, moderately productive and less productive. This study has revealed the Geospatial tools utilization along with the evaluation of the fisheries resources and creeks habitat risk zone mapping. It has also been identified that the geo-spatial technologies are highly beneficial to identify the areas of high environmental risk in Sindh Creeks. This has been clearly discovered from this study that creeks with high rugosity are more productive than the creeks with low levels of rugosity. The study area has the immense potential to boost the economy of Pakistan in terms of fish export, if geo-spatial techniques are implemented instead of conventional techniques.

Keywords: fish stock, geo-spatial, productivity analysis, risk

Procedia PDF Downloads 246

Authors: Hawraa Zbeeb, Hala Khalifeh, Mohamad Khalil, Francesca Storace, Francesca Baldini, Giulio Lupidi, Laura Vergani

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Sarcopoterium spinosum, a widely distributed spiny shrub belonging to the Rosaceae family, is rich in essential and beneficial constituents. In fact, S. spinosum fruits and roots are traditionally used as herbal medicine in the eastern Mediterranean landscape, and this shrub is mentioned as a medicinal plant in a large number of ethnobotanical surveys. Aqueous root extracts from S. spinosum are used by traditional medicinal practitioners for weight loss treatment of diabetes and pain. Moreover, the anti-diabetic activity of S. spinosum root extract has been reported in different studies, but the beneficial effects of aerial parts, especially fruits, have not been elucidated yet. The aim of the present study was to investigate the in vitro antioxidant and lipid-lowering properties of an ethanolic extract from S. spinosum fruits using both hepatic (FaO) and endothelial (HECV) cells in an attempt to evaluate its possible employment as a nutraceutical supplement. First of all, in vitro spectrophotometric assays were employed to characterize the extract. The total phenol content (TPC) was evaluated by Folin–Ciocalteu spectrophotometric method and the radical scavenging activity was tested by 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2, 2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) assays. After that, the beneficial effects of the extract were tested on cells. FaO cells treated for 3 hours with 0.75 mM oleate/palmitate mix (1:2 molar ratio) mimic in vitro a moderate hepato-steatosis. HECV cells exposed for 1 hour to 100 µM H₂O₂ mimic an oxidative insult leading to oxidative stress conditions. After the metabolic and oxidative insult, both cell lines were treated with increasing concentrations of the S. spinosum extract (1, 10, 25 µg/mL) for 24 hours. The results showed the S. spinosum ethanolic extract is rather rich in phenols (TPC of 18.6 mgGAE/g dry extracts). Moreover, the extract showed a good scavenging ability in vitro (IC₅₀ 15.9 µg/ml and 10.9 µg/ml measured by DPPH and ABTS assays, respectively). When the extract was tested on cells, the results showed that it could ameliorate some markers of cell dysfunction. The three concentrations of the extract led to a significant decrease in the intracellular triglyceride (TG) content in steatotic FaO cells measured by spectrophotometric assay. On the other hand, HECV cells treated with increasing concentrations of the extract did not result in a significant decrease in both lipid peroxidation measured by the Thiobarbituric Acid Reactive Substances (TBARS) assay, and in reactive oxygen species (ROS) production measured by fluorometric analysis after DCF staining. Interestingly, the ethanolic extract was able to accelerate the wound repair of confluent HECV cells with respect to H₂O₂-insulted cells as measured by T-scratch assay. Taken together, these results seem to indicate that the ethanol extract from S. spinosum fruits is rich in phenol compounds and plays considerable lipid-lowering activity in vitro on steatotic hepatocytes and accelerates wound healing repair on endothelial cells. In light of that, the ethanolic extract from S. spinosum fruits could be a potential candidate for nutraceutical applications.

Keywords: antioxidant activity, ethanolic extract, lipid-lowering activity, phenolic compounds, Sarcopoterium spinosum fruits

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Authors: Pomozova Kseniia, Gorlachev Gennadiy, Chernyaev Aleksandr, Golanov Andrey

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In clinical practice, and especially in stereotactic radiosurgery planning, the significance of diffusion-weighted imaging (DWI) is growing. This makes the existence of software capable of quickly processing and reliably visualizing diffusion data, as well as equipped with tools for their analysis in terms of different tasks. We are developing the «MRDiffusionImaging» software on the standard C++ language. The subject part has been moved to separate class libraries and can be used on various platforms. The user interface is Windows WPF (Windows Presentation Foundation), which is a technology for managing Windows applications with access to all components of the .NET 5 or .NET Framework platform ecosystem. One of the important features is the use of a declarative markup language, XAML (eXtensible Application Markup Language), with which you can conveniently create, initialize and set properties of objects with hierarchical relationships. Graphics are generated using the DirectX environment. The MRDiffusionImaging software package has been implemented for processing diffusion magnetic resonance imaging (dMRI), which allows loading and viewing images sorted by series. An algorithm for "masking" dMRI series based on T2-weighted images was developed using a deformable surface model to exclude tissues that are not related to the area of interest from the analysis. An algorithm of distortion correction using deformable image registration based on autocorrelation of local structure has been developed. Maximum voxel dimension was 1,03 ± 0,12 mm. In an elementary brain's volume, the diffusion tensor is geometrically interpreted using an ellipsoid, which is an isosurface of the probability density of a molecule's diffusion. For the first time, non-parametric intensity distributions, neighborhood correlations, and inhomogeneities are combined in one segmentation of white matter (WM), grey matter (GM), and cerebrospinal fluid (CSF) algorithm. A tool for calculating the coefficient of average diffusion and fractional anisotropy has been created, on the basis of which it is possible to build quantitative maps for solving various clinical problems. Functionality has been created that allows clustering and segmenting images to individualize the clinical volume of radiation treatment and further assess the response (Median Dice Score = 0.963 ± 0,137). White matter tracts of the brain were visualized using two algorithms: deterministic (fiber assignment by continuous tracking) and probabilistic using the Hough transform. The proposed algorithms test candidate curves in the voxel, assigning to each one a score computed from the diffusion data, and then selects the curves with the highest scores as the potential anatomical connections. White matter fibers were visualized using a Hough transform tractography algorithm. In the context of functional radiosurgery, it is possible to reduce the irradiation volume of the internal capsule receiving 12 Gy from 0,402 cc to 0,254 cc. The «MRDiffusionImaging» will improve the efficiency and accuracy of diagnostics and stereotactic radiotherapy of intracranial pathology. We develop software with integrated, intuitive support for processing, analysis, and inclusion in the process of radiotherapy planning and evaluating its results.

Keywords: diffusion-weighted imaging, medical imaging, stereotactic radiosurgery, tractography

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Authors: Jennifer Buz, Alvin Spivey

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The most common greenhouse gas emitted through human activities, carbon dioxide (CO2), is naturally consumed by plants during photosynthesis. This process is actively being monetized by companies wishing to offset their carbon dioxide emissions. For example, companies are now able to purchase protections for vegetated land due-to-be clear cut or purchase barren land for reforestation. Therefore, by actively preventing the destruction/decay of plant matter or by introducing more plant matter (reforestation), a company can theoretically offset some of their emissions. One of the biggest issues in the carbon credit market is validating and verifying carbon offsets. There is a need for a system that can accurately and frequently ensure that the areas sold for carbon credits have the vegetation mass (and therefore for carbon offset capability) they claim. Traditional techniques for measuring vegetation mass and determining health are costly and require many person-hours. Orbital Sidekick offers an alternative approach that accurately quantifies carbon mass and assesses vegetation health through satellite hyperspectral imagery, a technique which enables us to remotely identify material composition (including plant species) and condition (e.g., health and growth stage). How much carbon a plant is capable of storing ultimately is tied to many factors, including material density (primarily species-dependent), plant size, and health (trees that are actively decaying are not effectively storing carbon). All of these factors are capable of being observed through satellite hyperspectral imagery. This abstract focuses on speciation. To build a species classification model, we matched pixels in our remote sensing imagery to plants on the ground for which we know the species. To accomplish this, we collaborated with the researchers at the Teakettle Experimental Forest. Our remote sensing data comes from our airborne “Kato” sensor, which flew over the study area and acquired hyperspectral imagery (400-2500 nm, 472 bands) at ~0.5 m/pixel resolution. Coverage of the entire teakettle experimental forest required capturing dozens of individual hyperspectral images. In order to combine these images into a mosaic, we accounted for potential variations of atmospheric conditions throughout the data collection. To do this, we ran an open source atmospheric correction routine called ISOFIT1 (Imaging Spectrometer Optiman FITting), which converted all of our remote sensing data from radiance to reflectance. A database of reflectance spectra for each of the tree species within the study area was acquired using the Teakettle stem map and the geo-referenced hyperspectral images. We found that a wide variety of machine learning classifiers were able to identify the species within our images with high (>95%) accuracy. For the most robust quantification of carbon mass and the best assessment of the health of a vegetated area, speciation is critical. Through the use of high resolution hyperspectral data, ground-truth databases, and complex analytical techniques, we are able to determine the species present within a pixel to a high degree of accuracy. These species identifications will feed directly into our carbon mass model.

Keywords: hyperspectral, satellite, carbon, imagery, python, machine learning, speciation

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Authors: Sagar Saroha, Sawan S. Sinha, Sunil Lakshmipathy

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Reynolds averaged Navier-Stokes (RANS) model is the popular computational tool for prediction of turbulent flows. Being computationally less expensive as compared to direct numerical simulation (DNS), RANS has received wide acceptance in industry and research community as well. However, for high Reynolds number flows, the traditional RANS approach based on the Boussinesq hypothesis is incapacitated to capture all the essential flow characteristics, and thus, its performance is restricted in high Reynolds number flows of practical interest. RANS performance turns out to be inadequate in regimes like flow over curved surfaces, flows with rapid changes in the mean strain rate, duct flows involving secondary streamlines and three-dimensional separated flows. In the recent decade, partially averaged Navier-Stokes (PANS) methodology has gained acceptability among seamless bridging methods of turbulence- placed between DNS and RANS. PANS methodology, being a scale resolving bridging method, is inherently more suitable than RANS for simulating turbulent flows. The superior ability of PANS method has been demonstrated for some cases like swirling flows, high-speed mixing environment, and high Reynolds number turbulent flows. In our work, we intend to evaluate PANS in case of separated turbulent flows past bluff bodies -which is of broad aerodynamic research and industrial application. PANS equations, being derived from base RANS, continue to inherit the inadequacies from the parent RANS model based on linear eddy-viscosity model (LEVM) closure. To enhance PANS’ capabilities for simulating separated flows, the shortcomings of the LEVM closure need to be addressed. Inabilities of the LEVMs have inspired the development of non-linear eddy viscosity models (NLEVM). To explore the potential improvement in PANS performance, in our study we evaluate the PANS behavior in conjugation with NLEVM. Our work can be categorized into three significant steps: (i) Extraction of PANS version of NLEVM from RANS model, (ii) testing the model in the homogeneous turbulence environment and (iii) application and evaluation of the model in the canonical case of separated non-homogeneous flow field (flow past prismatic bodies and bodies of revolution at high Reynolds number). PANS version of NLEVM shall be derived and implemented in OpenFOAM -an open source solver. Homogeneous flows evaluation will comprise the study of the influence of the PANS’ filter-width control parameter on the turbulent stresses; the homogeneous analysis performed over typical velocity fields and asymptotic analysis of Reynolds stress tensor. Non-homogeneous flow case will include the study of mean integrated quantities and various instantaneous flow field features including wake structures. Performance of PANS + NLEVM shall be compared against the LEVM based PANS and LEVM based RANS. This assessment will contribute to significant improvement of the predictive ability of the computational fluid dynamics (CFD) tools in massively separated turbulent flows past bluff bodies.

Keywords: bridging methods of turbulence, high Re-CFD, non-linear PANS, separated turbulent flows

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Authors: Prince Chukwuneme Enwereji, Annelien Van Rooyen

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In recent years, technology integration in education has transformed the learning landscape, particularly in online institutions. One technological advancement that has gained popularity is e-tutoring, which offers personalised academic support to students through online platforms. While e-tutoring has become well-known and has been adopted to promote collaborative learning, there are still students who do not use these services for various reasons. However, little attention has been given to understanding the perspectives of students who have not utilized these services. The research objectives include identifying the perceived benefits that non-e-tutoring students believe e-tutoring could offer, such as enhanced academic support, personalized learning experiences, and improved performance. Additionally, the study explored the potential drawbacks or concerns that non-e-tutoring students associate with e-tutoring, such as concerns about efficacy, a lack of face-to-face interaction, and platform accessibility. The study adopted a quantitative research approach with a descriptive design to gather and analyze data on non-e-tutoring students' perspectives. Online questionnaires were employed as the primary data collection method, allowing for the efficient collection of data from many participants. The collected data was analyzed using the Statistical Package for the Social Sciences (SPSS). Ethical concepts such as informed consent, anonymity of responses and protection of respondents against harm were maintained. Findings indicate that non-e-tutoring students perceive a sense of control over their own pace of learning, suggesting a preference for self-directed learning and the ability to tailor their educational experience to their individual needs and learning styles. They also exhibit high levels of motivation, believe in their ability to effectively participate in their studies and organize their academic work, and feel comfortable studying on their own without the help of e-tutors. However, non-e-tutoring students feel that e-tutors do not sufficiently address their academic needs and lack engagement. They also perceive a lack of clarity in the roles of e-tutors, leading to uncertainty about their responsibilities. In terms of communication, students feel overwhelmed by the volume of announcements and find repetitive information frustrating. Additionally, some students face challenges with their internet connection and associated cost, which can hinder their participation in online activities. Furthermore, non-e-tutoring students express a desire for interactions with their peers and a sense of belonging to a group or team. They value opportunities for collaboration, teamwork in their learning experience, the importance of fostering social interactions and creating a sense of community in online learning environments. This study recommended that students seek alternate support systems by reaching out to professors or academic advisors for guidance and clarification. Developing self-directed learning skills is essential, empowering students to take charge of their own learning through setting objectives, creating own study plans, and utilising resources. For HEIs, it was recommended that they should ensure that a variety of support services are available to cater to the needs of all students, including non-e-tutoring students. HEIs should also ensure easy access to online resources, promote a supportive community, and regularly evaluate and adapt their support techniques to meet students' changing requirements.

Keywords: online-tutor;, student support;, online education, educational practices, distance education

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Authors: Ma Chu Jui

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Culture plays a pivotal role in shaping societal responses to change and fostering adaptability. In the realm of healthcare provision, hospitals serve as dynamic settings molded by the cultural consciousness of healthcare professionals. This intricate interplay extends to their expectations of leadership, communication styles, and attitudes towards patient care. Recognizing the cultural inclinations of healthcare professionals becomes imperative in navigating this complex landscape. This study will utilize Hofstede's Value Survey Module 2013 (VSM 2013) as a comprehensive analytical tool. The targeted participants for this research are in-service nursing professionals with a tenure of at least three months, specifically employed in the nursing department of an Eastern hospital. This quantitative approach seeks to quantify diverse cultural tendencies among the targeted nursing professionals, elucidating not only abstract cultural concepts but also revealing their cultural inclinations across different dimensions. The study anticipates gathering between 400 to 500 responses, ensuring a robust dataset for a comprehensive analysis. The focused approach on nursing professionals within the Eastern hospital setting enhances the relevance and specificity of the cultural insights obtained. The research aims to contribute valuable knowledge to the understanding of cultural tendencies among in-service nursing personnel in the nursing department of this specific Eastern hospital. The VSM 2013 will be initially distributed to this specific group to collect responses, aiming to calculate scores on each of Hofstede's six cultural dimensions—Power Distance Index (PDI), Individualism vs. Collectivism (IDV), Uncertainty Avoidance Index (UAI), Masculinity vs. Femininity (MAS), Long-Term Orientation vs. Short-Term Normative Orientation (LTO), and Indulgence vs. Restraint (IVR). the study unveils a significant correlation between different cultural dimensions and healthcare professionals' tendencies in understanding leadership expectations through PDI, grasping behavioral patterns via IDV, acknowledging risk acceptance through UAI, and understanding their long-term and short-term behaviors through LTO. These tendencies extend to communication styles and attitudes towards patient care. These findings provide valuable insights into the nuanced interconnections between cultural factors and healthcare practices. Through a detailed analysis of the varying levels of these cultural dimensions, we gain a comprehensive understanding of the predominant inclinations among the majority of healthcare professionals. This nuanced perspective adds depth to our comprehension of how cultural values shape their approach to leadership, communication, and patient care, contributing to a more holistic understanding of the healthcare landscape. A profound comprehension of the cultural paradigms embraced by healthcare professionals holds transformative potential. Beyond a mere understanding, it acts as a catalyst for elevating the caliber of healthcare services. This heightened awareness fosters cohesive collaboration among healthcare teams, paving the way for the establishment of a unified healthcare ethos. By cultivating shared values, our study envisions a healthcare environment characterized by enhanced quality, improved teamwork, and ultimately, a more favorable and patient-centric healthcare landscape. In essence, our research underscores the critical role of cultural awareness in shaping the future of healthcare delivery.

Keywords: hofstede's cultural, cultural dimensions, cultural values in healthcare, cultural awareness in nursing

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Authors: Tatiana Butkova, Nikolai Kibrik, Kristina Malsagova, Alexander Izotov, Alexander Stepanov, Anna Kaysheva

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Relevance. The genetic risk of developing schizophrenia is determined by two factors: single nucleotide polymorphisms and gene copy number variations. The search for serological markers for early diagnosis of schizophrenia is driven by the fact that the first five years of the disease are accompanied by significant biological, psychological, and social changes. It is during this period that pathological processes are most amenable to correction. The aim of this study was to analyze single nucleotide polymorphisms (SNPs) that are hypothesized to potentially influence the onset and development of the endogenous process. Materials and Methods It was analyzed 73 single nucleotide polymorphism variants. The study included 48 patients undergoing inpatient treatment at "Psychiatric Clinical Hospital No. 1" in Moscow, comprising 23 females and 25 males. Inclusion criteria: - Patients aged 18 and above. - Diagnosis according to ICD-10: F20.0, F20.2, F20.8, F21.8, F25.1, F25.2. - Voluntary informed consent from patients. Exclusion criteria included: - The presence of concurrent somatic or neurological pathology, neuroinfections, epilepsy, organic central nervous system damage of any etiology, and regular use of medication. - Substance abuse and alcohol dependence. - Women who were pregnant or breastfeeding. Clinical and psychopathological assessment was complemented by psychometric evaluation using the PANSS scale at the beginning and end of treatment. The duration of observation during therapy was 4-6 weeks. Total DNA extraction was performed using QIAamp DNA. Blood samples were processed on Illumina HiScan and genotyped for 652,297 markers on the Infinium Global Chips Screening Array-24v2.0 using the IMPUTE2 program with parameters Ne=20,000 and k=90. Additional filtration was performed based on INFO>0.5 and genotype probability>0.5. Quality control of the obtained DNA was conducted using agarose gel electrophoresis, with each tested sample having a volume of 100 µL. Results. It was observed that several SNPs exhibited gender dependence. We identified groups of single nucleotide polymorphisms with a membership of 80% or more in either the female or male gender. These SNPs included rs2661319, rs2842030, rs4606, rs11868035, rs518147, rs5993883, and rs6269.Another noteworthy finding was the limited combination of SNPs sufficient to manifest clinical symptoms leading to hospitalization. Among all 48 patients, each of whom was analyzed for deviations in 73 SNPs, it was discovered that the combination of involved SNPs in the manifestation of pronounced clinical symptoms of schizophrenia was 19±3 out of 73 possible. In study, the frequency of occurrence of single nucleotide polymorphisms also varied. The most frequently observed SNPs were rs4849127 (in 90% of cases), rs1150226 (86%), rs1414334 (75%), rs10170310 (73%), rs2857657, and rs4436578 (71%). Conclusion. Thus, the results of this study provide additional evidence that these genes may be associated with the development of schizophrenia spectrum disorders. However, it's impossible cannot rule out the hypothesis that these polymorphisms may be in linkage disequilibrium with other functionally significant polymorphisms that may actually be involved in schizophrenia spectrum disorders. It has been shown that missense SNPs by themselves are likely not causative of the disease but are in strong linkage disequilibrium with non-functional SNPs that may indeed contribute to disease predisposition.

Keywords: gene polymorphisms, genotyping, single nucleotide polymorphisms, schizophrenia.

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Authors: Wissam Saliba, Mandy Nicholson

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Secretory carcinoma (SC) is a rare type of salivary gland cancer. It was first realized as a distinct type of malignancy in 2010and wasinitially termed “mammary analogue secretory carcinoma” because of similarities with secretory breast cancer. The name was later changed to SC. Most SCs originate in parotid glands, and most harbour a rare gene mutation: ETV6-NTRK3. This mutation is rare in common cancers and common in rare cancers; it is present in most secretory carcinomas. Disease outcomes for SC are usually described as favourable as many cases of SC are lowgrade (LG), and cancer growth is slow. In early stages, localized therapy is usually indicated (surgery and/or radiation). Despitea favourable prognosis, a sub-set of casescan be much more aggressive.These cases tend to be of high-grade(HG).HG casesare associated with a poorer prognosis.Management of such cases can be challenging due to limited evidence for effective systemic therapy options. This case report describes the clinical management of a 46-year-oldmale patient with a unique case of SC. He was initially diagnosed with a low/intermediate grade carcinoma of the left parotid gland in 2009; he was treated with surgery and adjuvant radiation. Surgical pathology favoured primary salivary adenocarcinoma, and 2 lymph nodes were positive for malignancy. SC was not yet realized as a distinct type of cancerat the time of diagnosis, and the pathology reportvalidated this gap by stating that the specimen lacked features of the defined types of salivary carcinoma.Slow-growing pulmonary nodules were identified in 2017. In 2020, approximately 11 years after the initial diagnosis, the patient presented with malignant pleural effusion. Pathology from a pleural biopsy was consistent with metastatic poorly differentiated cancer of likely parotid origin, likely mammary analogue secretory carcinoma. The specimen was sent for Next Generation Sequencing (NGS); ETV6-NTRK3 gene fusion was confirmed, and systemic therapy was initiated.One cycle ofcarboplatin/paclitaxel was given in June 2020. He was switched to Larotrectinib (NTRK inhibitor (NTRKi)) later that month. Larotrectinib continued for approximately 9 months, with discontinuation in March 2021 due to disease progression. A second-generation NTRKi (Selitrectinib) was accessed and prescribedthrough a single patient study. Selitrectinib was well tolerated. The patient experienced a complete radiological response within~4 months. Disease progression occurred once again in October 2021. Progression was slow, and Selitrectinib continuedwhile the medical team performed a thorough search for additional treatment options. In January 2022, a liver lesion biopsy was performed, and NGS showed an NTRKG623R solvent-front resistance mutation. Various treatment pathways were considered. The patient pursuedanother investigational NTRKi through a clinical trial, and Selitrectinib was discontinued in July 2022. Excellent performance status was maintained throughout the entire course of treatment.It can be concluded that NTRK inhibitors provided satisfactory treatment efficacy and tolerance for this patient with high-grade transformation and NTRK gene fusion cancer. In the future, more clinical research is needed on systemic treatment options for high-grade transformations in NTRK gene fusion SCs.

Keywords: secretory carcinoma, high-grade transformations, NTRK gene fusion, NTRK inhibitor

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Authors: Melanie Grier, Adam Murry

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Objective: Worldwide, Indigenous Peoples experience underemployment and poverty at disproportionately higher rates than non-Indigenous people, despite similar rates of employment seeking. Euro-colonial conquest and genocidal assimilation policies are implicated as perpetuating poverty, which research consistently links to health and wellbeing disparities. Many of the contributors to poverty, such as inadequate income and lack of access to medical care, can be directly or indirectly linked to underemployment. Calls have been made to prioritize Indigenous perspectives in Industrial-Organizational (I/O) psychology research, yet the literature on Indigenous employment remains scarce. What does exist is disciplinarily diverse, topically scattered, and lacking evidence of community-based participatory research (CBPR) practices, a research project approach which prioritizes community leadership, partnership, and betterment and reduces the potential for harm. Due to the harmful colonial legacy of extractive scientific inquiry "on" rather than "with" Indigenous groups, Indigenous leaders and research funding agencies advocate for academic researchers to adopt reparative research methodologies such as CBPR to be used when studying issues pertaining to Indigenous Peoples or individuals. However, the frequency and consistency of CBPR implementation within scholarly discourse are unknown. Therefore, this project’s goal is two-fold: (1) to understand what comprises CBPR in Indigenous research and (2) to determine if CBPR has been historically used in Indigenous employment research. Method: Using a systematic literature review process, sixteen articles about CBPR use with Indigenous groups were selected, and content was analyzed to identify key components comprising CBPR usage. An Indigenous CBPR components framework was constructed and subsequently utilized to analyze the Indigenous employment empirical literature. A similar systematic literature review process was followed to search for relevant empirical articles on Indigenous employment. A total of 120 articles were identified in six global regions: Australia, New Zealand, Canada, America, the Pacific Islands, and Greenland/Norway. Each empirical study was procedurally examined and coded for criteria inclusion using content analysis directives. Results: Analysis revealed that, in total, CBPR elements were used 14% of the time in Indigenous employment research. Most studies (n=69; 58%) neglected to mention using any CBPR components, while just two studies discussed implementing all sixteen (2%). The most significant determinant of overall CBPR use was community member partnership (CP) in the research process. Studies from New Zealand were most likely to use CBPR components, followed by Canada, Australia, and America. While CBPR use did increase slowly over time, meaningful temporal trends were not found. Further, CBPR use did not directly correspond with the total number of topical articles published that year. Conclusions: Community-initiated and engaged research approaches must be better utilized in employment studies involving Indigenous Peoples. Future research efforts must be particularly attentive to community-driven objectives and research protocols, emphasizing specific areas of concern relevant to the field of I/O psychology, such as organizational support, recruitment, and selection.

Keywords: community-based participatory research, content analysis, employment, indigenous research, international, reconciliation, recruitment, reparative research, selection, systematic literature review

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Authors: Ricardo Alberto Chiong Zevallos, Eduardo Moraes Rego Reis

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Pancreatic adenocarcinoma (PDAC) patients have a less than 10% 5-year survival rate. PDAC has no defined diagnostic and prognostic biomarkers. Gemcitabine is the first-line drug in PDAC and several other cancers. Long non-coding RNAs (lncRNAs) contribute to the tumorigenesis and are potential biomarkers for PDAC. Although lncRNAs aren’t translated into proteins, they have important functions. LncRNAs can decoy or recruit proteins from the epigenetic machinery, act as microRNA sponges, participate in protein translocation through different cellular compartments, and even promote chemoresistance. The chromatin remodeling enzyme EZH2 is a histone methyltransferase that catalyzes the methylation of histone 3 at lysine 27, silencing local expression. EZH2 is ambivalent, it can also activate gene expression independently of its histone methyltransferase activity. EZH2 is overexpressed in several cancers and interacts with lncRNAs, being recruited to a specific locus. EZH2 can be recruited to activate an oncogene or silence a tumor suppressor. The lncRNAs misregulation in cancer can result in the differential recruitment of EZH2 and in a distinct epigenetic landscape, promoting chemoresistance. The relevance of the EZH2-lncRNAs interaction to chemoresistant PDAC was assessed by Real Time quantitative PCR (RT-qPCR) and RNA Immunoprecipitation (RIP) experiments with naïve and gemcitabine-resistant PDAC cells. The expression of several lncRNAs and EZH2 gene targets was evaluated contrasting naïve and resistant cells. Selection of candidate genes was made by bioinformatic analysis and literature curation. Indeed, the resistant cell line showed higher expression of chemoresistant-associated lncRNAs and protein coding genes. RIP detected lncRNAs interacting with EZH2 with varying intensity levels in the cell lines. During RIP, the nuclear fraction of the cells was incubated with an antibody for EZH2 and with magnetic beads. The RNA precipitated with the beads-antibody-EZH2 complex was isolated and reverse transcribed. The presence of candidate lncRNAs was detected by RT-qPCR, and the enrichment was calculated relative to INPUT (total lysate control sample collected before RIP). The enrichment levels varied across the several lncRNAs and cell lines. The EZH2-lncRNA interaction might be responsible for the regulation of chemoresistance-associated genes in multiple cancers. The relevance of the lncRNA-EZH2 interaction to PDAC was assessed by siRNA knockdown of a lncRNA, followed by the analysis of the EZH2 target expression by RT-qPCR. The chromatin immunoprecipitation (ChIP) of EZH2 and H3K27me3 followed by RT-qPCR with primers for EZH2 targets also assess the specificity of the EZH2 recruitment by the lncRNA. This is the first report of the interaction of EZH2 and lncRNAs HOTTIP and PVT1 in chemoresistant PDAC. HOTTIP and PVT1 were described as promoting chemoresistance in several cancers, but the role of EZH2 is not clarified. For the first time, the lncRNA LINC01133 was detected in a chemoresistant cancer. The interaction of EZH2 with LINC02577, LINC00920, LINC00941, and LINC01559 have never been reported in any context. The novel lncRNAs-EZH2 interactions regulate chemoresistant-associated genes in PDAC and might be relevant to other cancers. Therapies targeting EZH2 alone weren’t successful, and a combinatorial approach also targeting the lncRNAs interacting with it might be key to overcome chemoresistance in several cancers.

Keywords: epigenetics, chemoresistance, long non-coding RNAs, pancreatic cancer, histone modification

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Authors: Adenike Adesanya, Nonthaphat Wong, Xiang-Yun Lan, Shea Ping Yip, Chien-Ling Huang

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Background: The most challenging mutation of the oncokinase BCR-ABL protein T315I, which is commonly known as the “gatekeeper” mutation and is notorious for its strong resistance to almost all tyrosine kinase inhibitors (TKIs), especially imatinib. Therefore, this study aims to identify T315I-dependent downstream microRNA (miRNA) pathways associated with drug resistance in chronic myeloid leukemia (CML) for prognostic and therapeutic purposes. Methods: T315I-carrying K562 cell clones (K562-T315I) were generated by the CRISPR-Cas9 system. Imatinib-treated K562-T315I cells were subjected to small RNA library preparation and next-generation sequencing. Putative lncRNA-miRNA-mRNA networks were analyzed with (i) DESeq2 to extract differentially expressed miRNAs, using Padj value of 0.05 as cut-off, (ii) STarMir to obtain potential miRNA response element (MRE) binding sites of selected miRNAs on lncRNA H19, (iii) miRDB, miRTarbase, and TargetScan to predict mRNA targets of selected miRNAs, (iv) IntaRNA to obtain putative interactions between H19 and the predicted mRNAs, (v) Cytoscape to visualize putative networks, and (vi) several pathway analysis platforms – Enrichr, PANTHER and ShinyGO for pathway enrichment analysis. Moreover, mitochondria isolation and transcript quantification were adopted to determine the new mechanism involved in T315I-mediated resistance of CML treatment. Results: Verification of the CRISPR-mediated mutagenesis with digital droplet PCR detected the mutation abundance of ≥80%. Further validation showed the viability of ≥90% by cell viability assay, and intense phosphorylated CRKL protein band being detected with no observable change for BCR-ABL and c-ABL protein expressions by Western blot. As reported by several investigations into hematological malignancies, we determined a 7-fold increase of H19 expression in K562-T315I cells. After imatinib treatment, a 9-fold increment was observed. DESeq2 revealed 171 miRNAs were differentially expressed K562-T315I, 112 out of these miRNAs were identified to have MRE binding regions on H19, and 26 out of the 112 miRNAs were significantly downregulated. Adopting the seed-sequence analysis of these identified miRNAs, we obtained 167 mRNAs. 6 hub miRNAs (hsa-let-7b-5p, hsa-let-7e-5p, hsa-miR-125a-5p, hsa-miR-129-5p, and hsa-miR-372-3p) and 25 predicted genes were identified after constructing hub miRNA-target gene network. These targets demonstrated putative interactions with H19 lncRNA and were mostly enriched in pathways related to cell proliferation, senescence, gene silencing, and pluripotency of stem cells. Further experimental findings have also shown the up-regulation of mitochondrial transcript and lncRNA MALAT1 contributing to the lncRNA-miRNA-mRNA networks induced by BCR-ABL T315I mutation. Conclusions: Our results have indicated that lncRNA-miRNA regulators play a crucial role not only in leukemogenesis but also in drug resistance, considering the significant dysregulation and interactions in the K562-T315I cell model generated by CRISPR-Cas9. In silico analysis has further shown that lncRNAs H19 and MALAT1 bear several complementary miRNA sites. This implies that they could serve as a sponge, hence sequestering the activity of the target miRNAs.

Keywords: chronic myeloid leukemia, imatinib resistance, lncRNA-miRNA-mRNA, T315I mutation

Procedia PDF Downloads 160

Authors: Ponchang Apollos Wuyep, Alice Njolke Mafe, Longchi Satkat Zacheaus, Dogun Ojochogu, Dabot Ayuba Yakubu

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Fungi causes huge losses when infections occur both in plants and animals. Synthetic Antifungal drugs are mostly very expensive and highly cytotoxic when taken. This study was aimed at determining the in vitro and in vivo antifungal activities of the leaves and stem extracts of Terminalia mantaly (Umbrella tree)H. Perrier on Aspergillus species in a bid to identify potential sources of cheap starting materials for the synthesis of new drugs to address the growing antimicrobial resistance. T. mantaly leave and stem powdered plant was extracted by fractionation using the method of solvent partition co-efficient in their graded form in the order n-hexane, Ethyl acetate, methanol and distilled water and phytochemical screening of each fraction revealed the presence of alkaloids, saponins, Tannins, flavonoids, carbohydrates, steroids, anthraquinones, cardiac glycosides and terpenoids in varying degrees. The Agar well diffusion technique was used to screen for antifungal activity of the fractions on clinical isolates of Aspergillus species (Aspergillus flavus and Aspergillus fumigatus). Minimum inhibitory concentration (MIC50) of the most active extracts was determined by the broth dilution method. The fractions test indicated a high antifungal activity with zones of inhibition ranging from 6 to 26 mm and 8 to 30mm (leave fractions) and 10mm to 34mm and 14mm to36mm (stem fractions) on A. flavus and A. fumigatus respectively. All the fractions indicated antifungal activity in a dose response relationship at concentrations of 62.5mg/ml, 125mg/ml, 250mg/ml and 500mg/ml. Better antifungal efficacy was shown by the Ethyl acetate, Hexane and Methanol fractions in the in vitro as the most potent fraction with MIC ranging from 62.5 to 125mg/ml. There was no statistically significant difference (P>0.05) in the potency of the Eight fractions from leave and stem (Hexane, Ethyl acetate, methanol and distilled water, antifungal (fluconazole), which served as positive control and 10% DMSO(Dimethyl Sulfoxide)which served as negative control. In the in vivo investigations, the ingestion technique was used for the infectious studies Female Drosophilla melanogaster(UAS-Diptericin)normal flies(positive control),infected and not treated flies (negative control) and infected flies with A. fumigatus and placed on normal diet, diet containing fractions(MSM and HSM each at concentrations of 10mg/ml 20mg/ml, 30mg/ml, 40mg/ml, 50mg/ml, 60mg/ml, 70mg/ml, 80mg/ml, 90mg/ml and 100mg/ml), diet containing control drugs(fluconazole as positive control)and infected flies on normal diet(negative control), the flies were observed for fifteen(15) days. Then the total mortality of flies was recorded each day. The results of the study reveals that the flies were susceptible to infection with A. fumigatus and responded to treatment with more effectiveness at 50mg/ml, 60mg/ml and 70mg/ml for both the Methanol and Hexane stem fractions. Therefore, the Methanol and Hexane stem fractions of T. mantaly contain therapeutically useful compounds, justifying the traditional use of this plant for the treatment of fungal infections.

Keywords: Terminalia mantaly, Aspergillus fumigatus, cytotoxic, Drosophila melanogaster, antifungal

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Authors: Nilki Upathissa, Torin Wirasinghe

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This solution addresses the challenges faced by traditional bookstores and the limitations of digital media, striking a balance between the tactile experience of printed books and the convenience of modern technology. The book exchange system offers a sustainable alternative, empowering users to access a diverse range of books while promoting community engagement. The user-friendly interfaces incorporated into the book exchange system ensure a seamless and enjoyable experience for users. Intuitive features for book management, search, and messaging facilitate effortless exchanges and interactions between users. By streamlining the process, the system encourages readers to explore new books aligned with their interests, enhancing the overall reading experience. Central to the system's success is the hybrid recommendation engine, which leverages advanced technologies such as Long Short-Term Memory (LSTM) models. By analyzing user input, the engine accurately predicts genre preferences, enabling personalized book recommendations. The hybrid approach integrates multiple technologies, including user interfaces, machine learning models, and recommendation algorithms, to ensure the accuracy and diversity of the recommendations. The evaluation of the book exchange system with the hybrid recommendation engine demonstrated exceptional performance across key metrics. The high accuracy score of 0.97 highlights the system's ability to provide relevant recommendations, enhancing users' chances of discovering books that resonate with their interests. The commendable precision, recall, and F1score scores further validate the system's efficacy in offering appropriate book suggestions. Additionally, the curve classifications substantiate the system's effectiveness in distinguishing positive and negative recommendations. This metric provides confidence in the system's ability to navigate the vast landscape of book choices and deliver recommendations that align with users' preferences. Furthermore, the implementation of this book exchange system with a hybrid recommendation engine has the potential to revolutionize the way readers interact with printed books. By facilitating book exchanges and providing personalized recommendations, the system encourages a sense of community and exploration within the reading community. Moreover, the emphasis on sustainability aligns with the growing global consciousness towards eco-friendly practices. With its robust technical approach and promising evaluation results, this solution paves the way for a more inclusive, accessible, and enjoyable reading experience for book lovers worldwide. In conclusion, the developed book exchange system with a hybrid recommendation engine represents a progressive solution to the challenges faced by traditional bookstores and the limitations of digital media. By promoting sustainability, widening access to printed books, and fostering engagement with reading, this system addresses the evolving needs of book enthusiasts. The integration of user-friendly interfaces, advanced machine learning models, and recommendation algorithms ensure accurate and diverse book recommendations, enriching the reading experience for users.

Keywords: recommendation systems, hybrid recommendation systems, machine learning, data science, long short-term memory, recurrent neural network

Procedia PDF Downloads 97