Search results for: myostatin signaling pathway
Commenced in January 2007
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Edition: International
Paper Count: 944

Search results for: myostatin signaling pathway

74 Sustainable Production of Pharmaceutical Compounds Using Plant Cell Culture

Authors: David A. Ullisch, Yantree D. Sankar-Thomas, Stefan Wilke, Thomas Selge, Matthias Pump, Thomas Leibold, Kai Schütte, Gilbert Gorr

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Plants have been considered as a source of natural substances for ages. Secondary metabolites from plants are utilized especially in medical applications but are more and more interesting as cosmetical ingredients and in the field of nutraceuticals. However, supply of compounds from natural harvest can be limited by numerous factors i.e. endangered species, low product content, climate impacts and cost intensive extraction. Especially in the pharmaceutical industry the ability to provide sufficient amounts of product and high quality are additional requirements which in some cases are difficult to fulfill by plant harvest. Whereas in many cases the complexity of secondary metabolites precludes chemical synthesis on a reasonable commercial basis, plant cells contain the biosynthetic pathway – a natural chemical factory – for a given compound. A promising approach for the sustainable production of natural products can be plant cell fermentation (PCF®). A thoroughly accomplished development process comprises the identification of a high producing cell line, optimization of growth and production conditions, the development of a robust and reliable production process and its scale-up. In order to address persistent, long lasting production, development of cryopreservation protocols and generation of working cell banks is another important requirement to be considered. So far the most prominent example using a PCF® process is the production of the anticancer compound paclitaxel. To demonstrate the power of plant suspension cultures here we present three case studies: 1) For more than 17 years Phyton produces paclitaxel at industrial scale i.e. up to 75,000 L in scale. With 60 g/kg dw this fully controlled process which is applied according to GMP results in outstanding high yields. 2) Thapsigargin is another anticancer compound which is currently isolated from seeds of Thapsia garganica. Thapsigargin is a powerful cytotoxin – a SERCA inhibitor – and the precursor for the derivative ADT, the key ingredient of the investigational prodrug Mipsagargin (G-202) which is in several clinical trials. Phyton successfully generated plant cell lines capable to express this compound. Here we present data about the screening for high producing cell lines. 3) The third case study covers ingenol-3-mebutate. This compound is found in the milky sap of the intact plants of the Euphorbiacae family at very low concentrations. Ingenol-3-mebutate is used in Picato® which is approved against actinic keratosis. Generation of cell lines expressing significant amounts of ingenol-3-mebutate is another example underlining the strength of plant cell culture. The authors gratefully acknowledge Inspyr Therapeutics for funding.

Keywords: Ingenol-3-mebutate, plant cell culture, sustainability, thapsigargin

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73 Vertical Village Buildings as Sustainable Strategy to Re-Attract Mega-Cities in Developing Countries

Authors: M. J. Eichner, Y. S. Sarhan

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Overall study purpose has been the evaluation of ‘Vertical Villages’ as a new sustainable building typology, reducing significantly negative impacts of rapid urbanization processes in third world capital cities. Commonly in fast-growing cities, housing and job supply, educational and recreational opportunities, as well as public transportation infrastructure, are not accommodating rapid population growth, exposing people to high noise and emission polluted living environments with low-quality neighborhoods and a lack of recreational areas. Like many others, Egypt’s capital city Cairo, according to the UN facing annual population growth rates of up to 428.000 people, is struggling to address the general deterioration of urban living conditions. New settlements typologies and urban reconstruction approach hardly follow sustainable urbanization principles or socio-ecologic urbanization models with severe effects not only for inhabitants but also for the local environment and global climate. The authors prove that ‘Vertical Village’ buildings can offer a sustainable solution for increasing urban density with at the same time improving the living quality and urban environment significantly. Inserting them within high-density urban fabrics the ecologic and socio-cultural conditions of low-quality neighborhoods can be transformed towards districts, considering all needs of sustainable and social urban life. This study analyzes existing building typologies in Cairo’s «low quality - high density» districts Ard el Lewa, Dokki and Mohandesen according to benchmarks for sustainable residential buildings, identifying major problems and deficits. In 3 case study design projects, the sustainable transformation potential through ‘Vertical Village’ buildings are laid out and comparative studies show the improvement of the urban microclimate, safety, social diversity, sense of community, aesthetics, privacy, efficiency, healthiness and accessibility. The main result of the paper is that the disadvantages of density and overpopulation in developing countries can be converted with ‘Vertical Village’ buildings into advantages, achieving attractive and environmentally friendly living environments with multiple synergies. The paper is documenting based on scientific criteria that mixed-use vertical building structures, designed according to sustainable principles of low rise housing, can serve as an alternative to convert «low quality - high density» districts in megacities, opening a pathway for governments to achieve sustainable urban transformation goals. Neglected informal urban districts, home to millions of the poorer population groups, can be converted into healthier living and working environments.

Keywords: sustainable, architecture, urbanization, urban transformation, vertical village

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72 Metabolomics Fingerprinting Analysis of Melastoma malabathricum L. Leaf of Geographical Variation Using HPLC-DAD Combined with Chemometric Tools

Authors: Dian Mayasari, Yosi Bayu Murti, Sylvia Utami Tunjung Pratiwi, Sudarsono

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Melastoma malabathricum L. is an Indo-Pacific herb that has been traditionally used to treat several ailments such as wounds, dysentery, diarrhea, toothache, and diabetes. This plant is common across tropical Indo-Pacific archipelagos and is tolerant of a range of soils, from low-lying areas subject to saltwater inundation to the salt-free conditions of mountain slopes. How the soil and environmental variation influences secondary metabolite production in the herb, and an understanding of the plant’s utility as traditional medicine, remain largely unknown and unexplored. The objective of this study is to evaluate the variability of the metabolic profiles of M. malabathricum L. across its geographic distribution. By employing high-performance liquid chromatography-diode array detector (HPLC-DAD), a highly established, simple, sensitive, and reliable method was employed for establishing the chemical fingerprints of 72 samples of M. malabathricum L. leaves from various geographical locations in Indonesia. Specimens collected from six terrestrial and archipelago regions of Indonesia were analyzed by HPLC to generate chromatogram peak profiles that could be compared across each region. Data corresponding to the common peak areas of HPLC chromatographic fingerprint were analyzed by hierarchical component analysis (HCA) and principal component analysis (PCA) to extract information on the most significant variables contributing to characterization and classification of analyzed samples data. Principal component values were identified as PC1 and PC2 with 41.14% and 19.32%, respectively. Based on variety and origin, the high-performance liquid chromatography method validated the chemical fingerprint results used to screen the in vitro antioxidant activity of M. malabathricum L. The result shows that the developed method has potential values for the quality of similar M. malabathrium L. samples. These findings provide a pathway for the development and utilization of references for the identification of M. malabathricum L. Our results indicate the importance of considering geographic distribution during field-collection efforts as they demonstrate regional metabolic variation in secondary metabolites of M. malabathricum L., as illustrated by HPLC chromatogram peaks and their antioxidant activities. The results also confirm the utility of this simple approach to a rapid evaluation of metabolic variation between plants and their potential ethnobotanical properties, potentially due to the environments from whence they were collected. This information will facilitate the optimization of growth conditions to suit particular medicinal qualities.

Keywords: fingerprint, high performance liquid chromatography, Melastoma malabathricum l., metabolic profiles, principal component analysis

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71 Rediscovering English for Academic Purposes in the Context of the UN’s Sustainable Developmental Goals

Authors: Sally Abu Sabaa, Lindsey Gutt

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In an attempt to use education as a way of raising a socially responsible and engaged global citizen, the YU-Bridge program, the largest and fastest pathway program of its kind in North America, has embarked on the journey of integrating general themes from the UN’s sustainable developmental goals (SDGs) in its English for Academic Purposes (EAP) curriculum. The purpose of this initiative was to redefine the general philosophy of education in the middle of a pandemic and align with York University’s University Academic Plan that was released in summer 2020 framed around the SDGs. The YUB program attracts international students from all over the world but mainly from China, and its goal is to enable students to achieve the minimum language requirement to join their undergraduate courses at York University. However, along with measuring outcomes, objectives, and the students’ GPA, instructors and academics are always seeking innovation of the YUB curriculum to adapt to the ever growing challenges of academics in the university context, in order to focus more on subject matter that students will be exposed to in their undergraduate studies. However, with the sudden change that has happened globally with the advance of the COVID-19 pandemic, and other natural disasters like the increase in forest fires and floods, rethinking the philosophy and goal of education was a must. Accordingly, the SDGs became the solid pillars upon which we, academics and administrators of the program, could build a new curriculum and shift our perspective from simply ESL education to education with moral and ethical goals. The preliminary implementation of this initiative was supported by an institutional-wide consultation with EAP instructors who have diverse experiences, disciplines, and interests. Along with brainstorming sessions and mini-pilot projects preceding the integration of the SDGs in the YUB-EAP curriculum, those meetings led to creating a general outline of a curriculum and an assessment framework that has the SDGs at its core with the medium of ESL used for language instruction. Accordingly, a community of knowledge exchange was spontaneously created and facilitated by instructors. This has led to knowledge, resources, and teaching pedagogies being shared and examined further. In addition, experiences and reactions of students are being shared, leading to constructive discussions about opportunities and challenges with the integration of the SDGs. The discussions have branched out to discussions about cultural and political barriers along with a thirst for knowledge and engagement, which has resulted in increased engagement not only on the part of the students but the instructors as well. Later in the program, two surveys will be conducted: one for the students and one for the instructors to measure the level of engagement of each in this initiative as well as to elicit suggestions for further development. This paper will describe this fundamental step into using ESL methodology as a mode of disseminating essential ethical and socially correct knowledge for all learners in the 21st Century, the students’ reactions, and the teachers’ involvement and reflections.

Keywords: EAP, curriculum, education, global citizen

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70 The Molecular Mechanism of Vacuolar Function in Yeast Cell Homeostasis

Authors: Chang-Hui Shen, Paulina Konarzewska

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Cell homeostasis is regulated by vacuolar activity and it has been shown that lipid composition of the vacuole plays an important role in vacuolar function. The major phosphoinositide species present in the vacuolar membrane include phosphatidylinositol 3,5-biphosphate (PI(3,5)P₂) which is generated from PI(3)P controlled by Fab1p. Deletion of FAB1 gene reduce the synthesis of PI(3,5)P₂ and thus result in enlarged or fragmented vacuoles, with neutral vacuolar pH due to reduced vacuolar H⁺-ATPase activity. These mutants also exhibited poor growth at high extracellular pH and in the presence of CaCl₂. Conversely, VPS34 regulates the synthesis of PI(3)P from phosphatidylinositol (PI), and the lack of Vps34p results in the reduction of vacuolar activity. Although the cellular observations are clear, it is still unknown about the molecular mechanism between the phospholipid biosynthesis pathway and vacuolar activity. Since both VPS34 and FAB1 are important in vacuolar activity, we hypothesize that the molecular mechanism of vacuolar function might be regulated by the transcriptional regulators of phospholipid biosynthesis. In this study, we study the role of the major phospholipid biosynthesis transcription factor, INO2, in the regulation of vacuolar activity. We first performed qRT-PCR to examine the effect of Ino2p on the expression of VPS34 and FAB1. Our results showed that VPS34 was upregulated in the presence of inositol for both WT and ino2Δ cells. However, FAB1 was only upregulated significantly in ino2Δ cells. This indicated that Ino2p might be the negative regulator for FAB1 expression. Next, growth sensitivity experiment showed that WT, vma3Δ, and ino2Δ grew well in growth medium buffered to pH 5.5 containing 10 mM CaCl₂. As cells were switched to growth medium buffered to pH 7 containing CaCl₂ WT, ino2Δ and opi1Δ showed growth reduction, whereas vma3Δ was completely nonviable. As the concentration of CaCl₂ was increased to 60 mM, ino2Δ cells showed moderate growth reduction compared to WT. This result suggests that ino2Δ cells have better vacuolar activity. Microscopic analysis and vacuolar acidification were employed to further elucidate the importance of INO2 in vacuolar homeostasis. Analysis of vacuolar morphology indicated that WT and vma3Δ cells displayed vacuoles that occupied a small area of the cell when grown in media buffered to pH 5.5. Whereas, ino2Δ displayed fragmented vacuoles. On the other hand, all strains grown in media buffered to pH 7, exhibited enlarged vacuoles that occupied most of the cell’s surface. This indicated that the presence of INO2 may play negative effect in vacuolar morphology when cells are grown in media buffered to pH 5.5. Furthermore, vacuolar acidification assay showed that only vma3Δ cells displayed notably less acidic vacuoles as cells were grown in media buffered to pH 5.5 and pH 7. Whereas, ino2Δ cells displayed more acidic pH compared to WT at pH7. Taken together, our results demonstrated the molecular mechanism of the vacuolar activity regulated by the phospholipid biosynthesis transcription factors Ino2p. Ino2p negatively regulates vacuolar activity through the expression of FAB1.

Keywords: vacuole, phospholipid, homeostasis, Ino2p, FAB1

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69 Carbon Capture and Storage by Continuous Production of CO₂ Hydrates Using a Network Mixing Technology

Authors: João Costa, Francisco Albuquerque, Ricardo J. Santos, Madalena M. Dias, José Carlos B. Lopes, Marcelo Costa

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Nowadays, it is well recognized that carbon dioxide emissions, together with other greenhouse gases, are responsible for the dramatic climate changes that have been occurring over the past decades. Gas hydrates are currently seen as a promising and disruptive set of materials that can be used as a basis for developing new technologies for CO₂ capture and storage. Its potential as a clean and safe pathway for CCS is tremendous since it requires only water and gas to be mixed under favorable temperatures and mild high pressures. However, the hydrates formation process is highly exothermic; it releases about 2 MJ per kilogram of CO₂, and it only occurs in a narrow window of operational temperatures (0 - 10 °C) and pressures (15 to 40 bar). Efficient continuous hydrate production at a specific temperature range necessitates high heat transfer rates in mixing processes. Past technologies often struggled to meet this requirement, resulting in low productivity or extended mixing/contact times due to inadequate heat transfer rates, which consistently posed a limitation. Consequently, there is a need for more effective continuous hydrate production technologies in industrial applications. In this work, a network mixing continuous production technology has been shown to be viable for producing CO₂ hydrates. The structured mixer used throughout this work consists of a network of unit cells comprising mixing chambers interconnected by transport channels. These mixing features result in enhanced heat and mass transfer rates and high interfacial surface area. The mixer capacity emerges from the fact that, under proper hydrodynamic conditions, the flow inside the mixing chambers becomes fully chaotic and self-sustained oscillatory flow, inducing intense local laminar mixing. The device presents specific heat transfer rates ranging from 107 to 108 W⋅m⁻³⋅K⁻¹. A laboratory scale pilot installation was built using a device capable of continuously capturing 1 kg⋅h⁻¹ of CO₂, in an aqueous slurry of up to 20% in mass. The strong mixing intensity has proven to be sufficient to enhance dissolution and initiate hydrate crystallization without the need for external seeding mechanisms and to achieve, at the device outlet, conversions of 99% in CO₂. CO₂ dissolution experiments revealed that the overall liquid mass transfer coefficient is orders of magnitude larger than in similar devices with the same purpose, ranging from 1 000 to 12 000 h⁻¹. The present technology has shown itself to be capable of continuously producing CO₂ hydrates. Furthermore, the modular characteristics of the technology, where scalability is straightforward, underline the potential development of a modular hydrate-based CO₂ capture process for large-scale applications.

Keywords: network, mixing, hydrates, continuous process, carbon dioxide

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68 Dys-Regulation of Immune and Inflammatory Response in in vitro Fertilization Implantation Failure Patients under Ovarian Stimulation

Authors: Amruta D. S. Pathare, Indira Hinduja, Kusum Zaveri

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Implantation failure (IF) even after the good-quality embryo transfer (ET) in the physiologically normal endometrium is the main obstacle in in vitro fertilization (IVF). Various microarray studies have been performed worldwide to elucidate the genes requisite for endometrial receptivity. These studies have included the population based on different phases of menstrual cycle during natural cycle and stimulated cycle in normal fertile women. Additionally, the literature is also available in recurrent implantation failure patients versus oocyte donors in natural cycle. However, for the first time, we aim to study the genomics of endometrial receptivity in IF patients under controlled ovarian stimulation (COS) during which ET is generally practised in IVF. Endometrial gene expression profiling in IF patients (n=10) and oocyte donors (n=8) were compared during window of implantation under COS by whole genome microarray (using Illumina platform). Enrichment analysis of microarray data was performed to determine dys-regulated biological functions and pathways using Database for Annotation, Visualization and Integrated Discovery, v6.8 (DAVID). The enrichment mapping was performed with the help of Cytoscape software. Microarray results were validated by real-time PCR. Localization of genes related to immune response (Progestagen-Associated Endometrial Protein (PAEP), Leukaemia Inhibitory Factor (LIF), Interleukin-6 Signal Transducer (IL6ST) was detected by immunohistochemistry. The study revealed 418 genes downregulated and 519 genes upregulated in IF patients compared to healthy fertile controls. The gene ontology, pathway analysis and enrichment mapping revealed significant downregulation in activation and regulation of immune and inflammation response in IF patients under COS. The lower expression of Progestagen Associated Endometrial Protein (PAEP), Leukemia Inhibitory Factor (LIF) and Interleukin 6 Signal Transducer (IL6ST) in cases compared to controls by real time and immunohistochemistry suggests the functional importance of these genes. The study was proved useful to uncover the probable reason of implantation failure being imbalance of immune and inflammatory regulation in our group of subjects. Based on the present study findings, a panel of significant dysregulated genes related to immune and inflammatory pathways needs to be further substantiated in larger cohort in natural as well as stimulated cycle. Upon which these genes could be screened in IF patients during window of implantation (WOI) before going for embryo transfer or any other immunological treatment. This would help to estimate the regulation of specific immune response during WOI in a patient. The appropriate treatment of either activation of immune response or suppression of immune response can be then attempted in IF patients to enhance the receptivity of endometrium.

Keywords: endometrial receptivity, immune and inflammatory response, gene expression microarray, window of implantation

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67 Human Dental Pulp Stem Cells Attenuate Streptozotocin-Induced Parotid Gland Injury in Rats

Authors: Gehan ElAkabawy

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Background: Diabetes mellitus causes severe deteriorations of almost all the organs and systems of the body, as well as significant damage to the oral cavity. The oral changes are mainly related to salivary glands dysfunction characterized by hyposalivation and xerostomia, which significantly reduce diabetic patients’ quality of life. Human dental pulp stem cells represent a promising source for cell-based therapies, owing to their easy, minimally invasive surgical access, and high proliferative capacity. It was reported that the trophic support mediated by dental pulp stem cells can rescue the functional and structural alterations of damaged salivary glands. However, potential differentiation and paracrine effects of human dental pulp stem cells in diabetic-induced parotid gland damage have not been previously investigated. Our study aimed to investigate the therapeutic effects of intravenous transplantation of human dental pulp stem cells (hDPSCs) on parotid gland injury in a rat model of streptozotocin (STZ)-induced type 1 diabetes. Methods: Thirty Sprague-Dawley male rats were randomly categorised into three groups: control, diabetic (STZ), and transplanted (STZ+hDPSCs). hDPSCs or vehicle was injected into the tail vein 7 days after STZ injection. The fasting blood glucose levels were monitored weekly. A glucose tolerance test was performed, and the parotid gland weight, salivary flow rate, oxidative stress indices, parotid gland histology, and caspase-3, vascular endothelial growth factor (VEGF), and proliferating cell nuclear antigen (PCNA) expression in parotid tissues were assessed 28 days post-transplantation. Results: Transplantation of hDPSCs downregulated blood glucose, improved the salivary flow rate, and reduced oxidative stress. The cells migrated to, survived, and differentiated into acinar, ductal, and myoepithelial cells in the STZ-injured parotid gland. Moreover, they downregulated the expression of caspase-3 and upregulated the expression of VEGF and PCNA, likely exerting pro-angiogenetic and antiapoptotic effects and promoting endogenous regeneration. In addition, the transplanted cells enhanced the parotid nitric oxide (NO) -tetrahydrobiopterin (BH4) pathway. Conclusions: Our results show that hDPSCs can migrate to and survive within the STZ-injured parotid gland, where they prevent its functional and morphological damage by restoring normal glucose levels, differentiating into parotid cell populations, and stimulating paracrine-mediated regeneration. Thus, hDPSCs may have therapeutic potential in the treatment of diabetes-induced parotid gland injury.

Keywords: dental pulp stem cells, diabetes, streptozotocin, parotid gland

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66 The Toxic Effects of Kynurenine Metabolites on SH-SY5Y Neuroblastoma Cells

Authors: Susan Hall, Gary D. Grant, Catherine McDermott, Devinder Arora

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Introduction /Aim: The kynurenine pathway is thought to play an important role in the pathophysiology of numerous neurodegenerative diseases including depression, Alzheimer’s disease, and Parkinson’s disease. Numerous neuroactive compounds, including the neurotoxic 3-hydroxyanthranilic acid, 3-hydroxykynurenine and quinolinic acid and the neuroprotective kynurenic acid and picolinic acid, are produced through the metabolism of kynurenine and are thought to be the causative agents responsible for neurodegeneration. The toxicity of 3-hydroxykynurenine, 3-hydroxyanthranilic acid and quinolinic acid has been widely evaluated and demonstrated in primary cell cultures but to date only 3-hydroxykynurenine and 3-hydroxyanthranilic acid have been shown to cause toxicity in immortal tumour cells. The aim of this study was to evaluate the toxicity of kynurenine metabolites, both individually and in combination, on SH-SY5Y neuroblastoma cells after 24 and 72 h exposure in order to explore a cost-effective model to study their neurotoxic effects and potential protective agents. Methods: SH-SY5Y neuroblastoma cells were exposed to various concentrations of the neuroactive kynurenine metabolites, both individually and in combination, for 24 and 72 h, and viability was subsequently evaluated using the Resazurin (Alamar blue) proliferation assay. Furthermore, the effects of these compounds, alone and in combination, on specific death pathways including apoptosis, necrosis and free radical production was evaluated using various assays. Results: Consistent with literature, toxicity was shown with short-term 24-hour treatments at 1000 μM concentrations for both 3-hydroxykynurenine and 3-hydroxyanthranilic acid. Combinations of kynurenine metabolites showed modest toxicity towards SH-SY5Y neuroblastoma cells in a concentration-dependent manner. Specific cell death pathways, including apoptosis, necrosis and free radical production were shown to be increased after both 24 and 72 h exposure of SH-SY5Y neuroblastoma cells to 3-hydroxykynurenine and 3-hydroxyanthranilic acid and various combinations of neurotoxic kynurenine metabolites. Conclusion: It is well documented that neurotoxic kynurenine metabolites show toxicity towards primary human neurons in the nanomolar to low micromolar concentration range. Results show that the concentrations required to show significant cell death are in the range of 1000 µM for 3-hydroxykynurenine and 3-hydroxyanthranilic acid and toxicity of quinolinic acid towards SH-SY5Y was unable to be shown. This differs significantly from toxicities observed in primary human neurons. Combinations of the neurotoxic metabolites were shown to have modest toxicity towards these cells with increased toxicity and activation of cell death pathways observed after 72 h exposure. This study suggests that the 24 h model is unsuitable for use in neurotoxicity studies, however, the 72 h model better represents the observations of the studies using primary human neurons and may provide some benefit in providing a cost-effective model to assess possible protective agents against kynurenine metabolite toxicities.

Keywords: kynurenine metabolites, neurotoxicity, quinolinic acid, SH-SY5Y neuroblastoma

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65 Causes and Consequences of Intuitive Animal Communication: A Case Study at Panthera Africa

Authors: Cathrine Scharning Cornwall-Nyquist, David Rafael Vaz Fernandes

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Since its origins, mankind has been dreaming of communicating directly with other animals. Past civilizations interacted on different levels with other species and recognized them in their rituals and daily activities. However, recent scientific developments have limited the ability of humans to consider deeper levels of interaction beyond observation and/or physical behavior. In recent years, animal caretakers and facilities such as sanctuaries or rescue centers have been introducing new techniques based on intuition. Most of those initiatives are related to specific cases, such as the incapacity to understand an animal’s behavior. Respected organizations also include intuitive animal communication (IAC) sessions to follow up on past interventions with their animals. Despite the lack of credibility of this discipline, some animal caring structures have opted to integrate IAC into their daily routines and approaches to animal welfare. At this stage, animal communication will be generally defined as the ability of humans to communicate with animals on an intuitive level. The trend in the field remains to be explored. The lack of theory and previous research urges the scientific community to improve the description of the phenomenon and its consequences. Considering the current scenario, qualitative approaches may become a suitable pathway to explore this topic. The purpose of this case study is to explore the beliefs behind and the consequences of an approach based on intuitive animal communication techniques for Panthera Africa (PA), an ethical sanctuary located in South Africa. Due to their personal experience, the Sanctuary’s founders have developed a philosophy based on IAC while respecting the world's highest standards for big cat welfare. Their dual approach is reflected in their rescues, daily activities, and healing animals’ trauma. The case study's main research questions will be: (i) Why do they choose to apply IAC in their work? (ii) What consequences to their activities do IAC bring? (iii) What effects do IAC techniques bring in their interactions with the outside world? Data collection will be gathered on-site via: (i) Complete participation (field notes); (ii) Semi-structured interviews (audio transcriptions); (iii) Document analysis (internal procedures and policies); (iv) Audio-visual material (communication with third parties). The main researcher shall become an active member of the Sanctuary during a 30-day period and have full access to the site. Access to documents and audio-visual materials will be granted on a request basis. Interviews are expected to be held with PA founders and staff members and with IAC practitioners related to the facility. The information gathered shall enable the researcher to provide an extended description of the phenomenon and explore its internal and external consequences for Panthera Africa.

Keywords: animal welfare, intuitive animal communication, Panthera Africa, rescue

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64 Concussion: Clinical and Vocational Outcomes from Sport Related Mild Traumatic Brain Injury

Authors: Jack Nash, Chris Simpson, Holly Hurn, Ronel Terblanche, Alan Mistlin

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There is an increasing incidence of mild traumatic brain injury (mTBI) cases throughout sport and with this, a growing interest from governing bodies to ensure these are managed appropriately and player welfare is prioritised. The Berlin consensus statement on concussion in sport recommends a multidisciplinary approach when managing those patients who do not have full resolution of mTBI symptoms. There are as of yet no standardised guideline to follow in the treatment of complex cases mTBI in athletes. The aim of this project was to analyse the outcomes, both clinical and vocational, of all patients admitted to the mild Traumatic Brain Injury (mTBI) service at the UK’s Defence Military Rehabilitation Centre Headley Court between 1st June 2008 and 1st February 2017, as a result of a sport induced injury, and evaluate potential predictive indicators of outcome. Patients were identified from a database maintained by the mTBI service. Clinical and occupational outcomes were ascertained from medical and occupational employment records, recorded prospectively, at time of discharge from the mTBI service. Outcomes were graded based on the vocational independence scale (VIS) and clinical documentation at discharge. Predictive indicators including referral time, age at time of injury, previous mental health diagnosis and a financial claim in place at time of entry to service were assessed using logistic regression. 45 Patients were treated for sport-related mTBI during this time frame. Clinically 96% of patients had full resolution of their mTBI symptoms after input from the mTBI service. 51% of patients returned to work at their previous vocational level, 4% had ongoing mTBI symptoms, 22% had ongoing physical rehabilitation needs, 11% required mental health input and 11% required further vestibular rehabilitation. Neither age, time to referral, pre-existing mental health condition nor compensation seeking had a significant impact on either vocational or clinical outcome in this population. The vast majority of patients reviewed in the mTBI clinic had persistent symptoms which could not be managed in primary care. A consultant-led, multidisciplinary approach to the diagnosis and management of mTBI has resulted in excellent clinical outcomes in these complex cases. High levels of symptom resolution suggest that this referral and treatment pathway is successful and is a model which could be replicated in other organisations with consultant led input. Further understanding of both predictive and individual factors would allow clinicians to focus treatments on those who are most likely to develop long-term complications following mTBI. A consultant-led, multidisciplinary service ensures a large number of patients will have complete resolution of mTBI symptoms after sport-related mTBI. Further research is now required to ascertain the key predictive indicators of outcome following sport-related mTBI.

Keywords: brain injury, concussion, neurology, rehabilitation, sports injury

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63 Targeting Matrix Metalloprotease-9 to Reduce Coronary Artery Manifestations of Kawasaki’s Disease

Authors: Mohammadjavad Sotoudeheian, Navid Farahmandian

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Kawasaki disease (KD) is the primary cause of acquired pediatric heart disease as an acute vasculitis. In children with prolonged fever, rash, and inflammation of the mucosa KD must be considered as a clinical diagnosis. There is a persuasive suggestion of immune-mediated damage as the pathophysiologic cascade of KD. For example, the invasion of cytotoxic T-cells supports a viral etiology and the inflammasome of the innate immune system is a critical component in the vasculitis formation in KD. Animal models of KD propose the cytokine profiles, such as increased IL-1 and GM-CSF, which cause vascular damage. CRP and IFN-γ elevated expression and the upregulation of IL-6, and IL-10 production are also described in previous studies. Untreated KD is a critical risk factor for coronary artery diseases and myocardial infarction. Vascular damage may encompass amplified T-cell activity. SMAD3 is an essential molecule in down-regulating T-cells and increasing expression of FoxP3. It has a critical effect in the differentiation of regulatory T-cells. The discrepancy of regulatory T-cells and pro-inflammatory Th17 has been studied in acute coronary syndrome during KD. However in the coronary artery damaged lymphocytes and IgA plasma cells are seen at the lesion locations, the major immune cells in the coronary lesions are monocytes/macrophages and neutrophils. These cells secrete TNF-α, and activates matrix metalloprotease (MMP)-9, reducing the integrity of vessels and prompting patients to arise aneurysm. MMPs can break down the components of the extracellular matrix and assist immune cell movement. IVIG as an effective form of treatment clarified the role of the immune system, which may target pathogenic antigens and regulate cytokine production. Several reports have revealed that in the coronary arteries, high expression of MMP-9 in monocyte/macrophage results in pathologic cascades. Curcumin is a potent antioxidant and anti-inflammatory molecule. Curcumin decreases the production of reactive oxygen and nitrogen species and inhibits transcription factors like AP-1 and NF-κB. Curcumin also contains the characteristics of inhibitory effects on MMPs, especially MMP-9. The upregulation of MMP-9 is an important cellular response. Curcumin treatment caused a reverse effect and down-regulates MMP-9 gene expression which may fund the anti-inflammatory effect. Curcumin inhibits MMP-9 expression via PKC and AMPK-dependent pathways in Human monocytes cells. Elevated expression and activity of MMP-9 are correlated with advanced vascular lesions. AMPK controls lipid metabolism and oxidation, and protein synthesis. AMPK is also necessary for the MMP-9 activity and THP-1 cell adhesion to endothelial cells. Curcumin was shown to inhibit the activation of AMPKα. Compound C (AMPK inhibitor) inhibits MMP-9 expression level. Therefore, through inactivating AMPKs and PKC, curcumin decreases the MMP-9 level, which results in inhibiting monocyte/macrophage differentiation. Compound C also suppress the phosphorylation of three major classes of MAP kinase signaling, suggesting that curcumin may suppress MMP-9 level by inactivation of MAPK pathways. MAPK cascades are activated to induce the expression of MMP-9. Curcumin inhibits MAPKs phosphorylation, which contributes to the down-regulation of MMP-9. This study demonstrated that the potential inhibitory properties of curcumin over MMP-9 lead to a therapeutic strategy to reduce the risk of coronary artery involvement during KD.

Keywords: MMP-9, coronary artery aneurysm, Kawasaki’s disease, curcumin, AMPK, immune system, NF-κB, MAPK

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62 Theoretical Study on the Visible-Light-Induced Radical Coupling Reactions Mediated by Charge Transfer Complex

Authors: Lishuang Ma

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Charge transfer (CT) complex, also known as Electron donor-acceptor (EDA) complex, has received attentions increasingly in the field of synthetic chemistry community, due to the CT complex can absorb the visible light through the intermolecular charge transfer excited states, various of catalyst-free photochemical transformations under mild visible-light conditions. However, a number of fundamental questions are still ambiguous, such as the origin of visible light absorption, the photochemical and photophysical properties of the CT complex, as well as the detailed mechanism of the radical coupling pathways mediated by CT complex. Since these are critical factors for target-specific design and synthesis of more new-type CT complexes. To this end, theoretical investigations were performed in our group to answer these questions based on multiconfigurational perturbation theory. The photo-induced fluoroalkylation reactions are mediated by CT complexes, which are formed by the association of an acceptor of perfluoroalkyl halides RF−X (X = Br, I) and a suitable donor molecule such as β-naphtholate anion, were chosen as a paradigm example in this work. First, spectrum simulations were carried out by both CASPT2//CASSCF/PCM and TD-DFT/PCM methods. The computational results showed that the broadening spectra in visible light range (360-550nm) of the CT complexes originate from the 1(σπ*) excitation, accompanied by an intermolecular electron transfer, which was also found closely related to the aggregate states of the donor and acceptor. Moreover, from charge translocation analysis, the CT complex that showed larger charge transfer in the round state would exhibit smaller charge transfer in excited stated of 1(σπ*), causing blue shift relatively. Then, the excited-state potential energy surface (PES) was calculated at CASPT2//CASSCF(12,10)/ PCM level of theory to explore the photophysical properties of the CT complexes. The photo-induced C-X (X=I, Br) bond cleavage was found to occur in the triplet state, which is accessible through a fast intersystem crossing (ISC) process that is controlled by the strong spin-orbit coupling resulting from the heavy iodine and bromine atoms. Importantly, this rapid fragmentation process can compete and suppress the backward electron transfer (BET) event, facilitating the subsequent effective photochemical transformations. Finally, the reaction pathways of the radical coupling were also inspected, which showed that the radical chain propagation pathway could easy to accomplish with a small energy barrier no more than 3.0 kcal/mol, which is the key factor that promote the efficiency of the photochemical reactions induced by CT complexes. In conclusion, theoretical investigations were performed to explore the photophysical and photochemical properties of the CT complexes, as well as the mechanism of radical coupling reactions mediated by CT complex. The computational results and findings in this work can provide some critical insights into mechanism-based design for more new-type EDA complexes

Keywords: charge transfer complex, electron transfer, multiconfigurational perturbation theory, radical coupling

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61 Modeling Visual Memorability Assessment with Autoencoders Reveals Characteristics of Memorable Images

Authors: Elham Bagheri, Yalda Mohsenzadeh

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Image memorability refers to the phenomenon where certain images are more likely to be remembered by humans than others. It is a quantifiable and intrinsic attribute of an image. Understanding how visual perception and memory interact is important in both cognitive science and artificial intelligence. It reveals the complex processes that support human cognition and helps to improve machine learning algorithms by mimicking the brain's efficient data processing and storage mechanisms. To explore the computational underpinnings of image memorability, this study examines the relationship between an image's reconstruction error, distinctiveness in latent space, and its memorability score. A trained autoencoder is used to replicate human-like memorability assessment inspired by the visual memory game employed in memorability estimations. This study leverages a VGG-based autoencoder that is pre-trained on the vast ImageNet dataset, enabling it to recognize patterns and features that are common to a wide and diverse range of images. An empirical analysis is conducted using the MemCat dataset, which includes 10,000 images from five broad categories: animals, sports, food, landscapes, and vehicles, along with their corresponding memorability scores. The memorability score assigned to each image represents the probability of that image being remembered by participants after a single exposure. The autoencoder is finetuned for one epoch with a batch size of one, attempting to create a scenario similar to human memorability experiments where memorability is quantified by the likelihood of an image being remembered after being seen only once. The reconstruction error, which is quantified as the difference between the original and reconstructed images, serves as a measure of how well the autoencoder has learned to represent the data. The reconstruction error of each image, the error reduction, and its distinctiveness in latent space are calculated and correlated with the memorability score. Distinctiveness is measured as the Euclidean distance between each image's latent representation and its nearest neighbor within the autoencoder's latent space. Different structural and perceptual loss functions are considered to quantify the reconstruction error. The results indicate that there is a strong correlation between the reconstruction error and the distinctiveness of images and their memorability scores. This suggests that images with more unique distinct features that challenge the autoencoder's compressive capacities are inherently more memorable. There is also a negative correlation between the reduction in reconstruction error compared to the autoencoder pre-trained on ImageNet, which suggests that highly memorable images are harder to reconstruct, probably due to having features that are more difficult to learn by the autoencoder. These insights suggest a new pathway for evaluating image memorability, which could potentially impact industries reliant on visual content and mark a step forward in merging the fields of artificial intelligence and cognitive science. The current research opens avenues for utilizing neural representations as instruments for understanding and predicting visual memory.

Keywords: autoencoder, computational vision, image memorability, image reconstruction, memory retention, reconstruction error, visual perception

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60 Impact Analysis of a School-Based Oral Health Program in Brazil

Authors: Fabio L. Vieira, Micaelle F. C. Lemos, Luciano C. Lemos, Rafaela S. Oliveira, Ian A. Cunha

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Brazil has some challenges ahead related to population oral health, most of them associated with the need of expanding into the local level its promotion and prevention activities, offer equal access to services and promote changes in the lifestyle of the population. The program implemented an oral health initiative in public schools in the city of Salvador, Bahia. The mission was to improve oral health among students on primary and secondary education, from 2 to 15 years old, using the school as a pathway to increase access to healthcare. The main actions consisted of a team's visit to the schools with educational sessions for dental cavity prevention and individual assessment. The program incorporated a clinical surveillance component through a dental evaluation of every student searching for dental disease and caries, standardization of the dentists’ team to reach uniform classification on the assessments, and the use of an online platform to register data directly from the schools. Sequentially, the students with caries were referred for free clinical treatment on the program’s Health Centre. The primary purpose of this study was to analyze the effects and outcomes of this school-based oral health program. The study sample was composed by data of a period of 3 years - 2015 to 2017 - from 13 public schools on the suburb of the city of Salvador with a total number of assessments of 9,278 on this period. From the data collected the prevalence of children with decay on permanent teeth was chosen as the most reliable indicator. The prevalence was calculated for each one of the 13 schools using the number of children with 1 or more dental caries on permanent teeth divided by the total number of students assessed for school each year. Then the percentage change per year was calculated for each school. Some schools presented a higher variation on the total number of assessments in one of the three years, so for these, the percentage change calculation was done using the two years with less variation. The results show that 10 of the 13 schools presented significative improvements for the indicator of caries in permanent teeth. The mean for the number of students with caries percentage reduction on the 13 schools was 26.8%, and the median was 32.2% caries in permanent teeth institution. The highest percentage of improvement reached a decrease of 65.6% on the indicator. Three schools presented a rise in caries prevalence (8.9, 18.9 and 37.2% increase) that, on an initial analysis, seems to be explained with the students’ cohort rotation among other schools, as well as absenteeism on the treatment. In conclusion, the program shows a relevant impact on the reduction of caries in permanent teeth among students and the need for the continuity and expansion of this integrated healthcare approach. It has also been evident the significative of the articulation between health and educational systems representing a fundamental approach to improve healthcare access for children especially in scenarios such as presented in Brazil.

Keywords: primary care, public health, oral health, school-based oral health, data management

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59 Graphene Supported Nano Cerium Oxides Hybrid as an Electrocatalyst for Oxygen Reduction Reactions

Authors: Siba Soren, Purnendu Parhi

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Today, the world is facing a severe challenge due to depletion of traditional fossil fuels. Scientists across the globe are working for a solution that involves a dramatic shift to practical and environmentally sustainable energy sources. High-capacity energy systems, such as metal-air batteries, fuel cells, are highly desirable to meet the urgent requirement of sustainable energies. Among the fuel cells, Direct methanol fuel cells (DMFCs) are recognized as an ideal power source for mobile applications and have received considerable attention in recent past. In this advanced electrochemical energy conversion technologies, Oxygen Reduction Reaction (ORR) is of utmost importance. However, the poor kinetics of cathodic ORR in DMFCs significantly hampers their possibilities of commercialization. The oxygen is reduced in alkaline medium either through a 4-electron (equation i) or a 2-electron (equation ii) reduction pathway at the cathode ((i) O₂ + 2H₂O + 4e⁻ → 4OH⁻, (ii) O₂ + H₂O + 2e⁻ → OH⁻ + HO₂⁻ ). Due to sluggish ORR kinetics the ability to control the reduction of molecular oxygen electrocatalytically is still limited. The electrocatalytic ORR starts with adsorption of O₂ on the electrode surface followed by O–O bond activation/cleavage and oxide removal. The reaction further involves transfer of 4 electrons and 4 protons. The sluggish kinetics of ORR, on the one hand, demands high loading of precious metal-containing catalysts (e.g., Pt), which unfavorably increases the cost of these electrochemical energy conversion devices. Therefore, synthesis of active electrocatalyst with an increase in ORR performance is need of the hour. In the recent literature, there are many reports on transition metal oxide (TMO) based ORR catalysts for their high activity TMOs are also having drawbacks like low electrical conductivity, which seriously affects the electron transfer process during ORR. It was found that 2D graphene layer is having high electrical conductivity, large surface area, and excellent chemical stability, appeared to be an ultimate choice as support material to enhance the catalytic performance of bare metal oxide. g-C₃N₄ is also another candidate that has been used by the researcher for improving the ORR performance of metal oxides. This material provides more active reaction sites than other N containing carbon materials. Rare earth oxide like CeO₂ is also a good candidate for studying the ORR activity as the metal oxide not only possess unique electronic properties but also possess catalytically active sites. Here we will discuss the ORR performance (in alkaline medium) of N-rGO/C₃N₄ supported nano Cerium Oxides hybrid synthesized by microwave assisted Solvothermal method. These materials exhibit superior electrochemical stability and methanol tolerance capability to that of commercial Pt/C.

Keywords: oxygen reduction reaction, electrocatalyst, cerium oxide, graphene

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58 An Australian Tertiary Centre Experience of Complex Endovascular Aortic Repairs

Authors: Hansraj Bookun, Rachel Xuan, Angela Tan, Kejia Wang, Animesh Singla, David Kim, Christopher Loupos, Jim Iliopoulos

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Introduction: Complex endovascular aortic aneursymal repairs with fenestrated and branched endografts require customised devices to exclude the pathology while reducing morbidity and mortality, which was historically associated with open repair of complex aneurysms. Such endovascular procedures have predominantly been performed in a large volume dedicated tertiary centres. We present here our nine year multidisciplinary experience with this technology in an Australian tertiary centre. Method: This was a cross-sectional, single-centre observational study of 670 patients who had undergone complex endovascular aortic aneurysmal repairs with conventional endografts, fenestrated endografts, and iliac-branched devices from January 2010 to July 2019. Descriptive statistics were used to characterise our sample with regards to demographic and perioperative variables. Homogeneity of the sample was tested using multivariant regression, which did not identify any statistically significant confounding variables. Results: 670 patients of mean age 74, were included (592 males) and the comorbid burden was as follows: ischemic heart disease (55%), diabetes (18%), hypertension (90%), stage four or greater kidney impairment (8%) and current or ex-smoking (78%). The main indications for surgery were elective aneurysms (86%), symptomatic aneurysms (5%), and rupture aneurysms (5%). 106 patients (16%) underwent fenestrated or branched endograft repairs. The mean length of stay was 7.6 days. 2 patients experienced reactionary bleeds, 11 patients had access wound complications (6 lymph fistulae, 5 haematoms), 11 patients had cardiac complications (5 arrhythmias, 3 acute myocadial infarctions, 3 exacerbation of congestive cardiac failure), 10 patients had respiratory complications, 8 patients had renal impairment, 4 patients had gastrointestinal complications, 2 patients suffered from paraplegia, 1 major stroke, 1 minor stroke, and 1 acute brain syndrome. There were 4 vascular occlusions requiring further arterial surgery, 4 type I endoleaks, 4 type II endoleaks, 3 episodes of thromboembolism, and 2 patients who required further arterial operations in the setting of patient vessels. There were 9 unplanned returns to the theatre. Discussion: Our numbers of 10 years suggest that we are not a dedicated high volume centre focusing on aortic repairs. However, we have achieved significantly low complication rates. This can be attributed to our multidisciplinary approach with the intraoperative involvement of skilled interventional radiologists and vascular surgeons as well as postoperative protocols with particular attention to spinal cord protection. Additionally, we have a ratified perioperative pathway that involves multidisciplinary team discussions of patient-related factors and lesion-centered characteristics, which allows for holistic, patient-centered care.

Keywords: aneurysm, aortic, endovascular, fenestrated

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57 A 3d Intestine-On-Chip Model Allows Colonization with Commensal Bacteria to Study Host-Microbiota Interaction

Authors: Michelle Maurer, Antonia Last, Mark S. Gresnigt, Bernhard Hube, Alexander S. Mosig

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The intestinal epithelium forms an essential barrier to prevent translocation of microorganisms, toxins or other potentially harmful molecules into the bloodstream. In particular, dendritic cells of the intestinal epithelium orchestrate an adapted response of immune tolerance to commensals and immune defense against invading pathogens. Systemic inflammation is typically associated with a dysregulation of this adapted immune response and is accompanied by a disruption of the epithelial and endothelial gut barrier which enables dissemination of pathogens within the human body. To understand the pathophysiological mechanisms underlying the inflammation-associated gut barrier breakdown, it is crucial to elucidate the complex interplay of the host and the intestinal microbiome. A microfluidically perfused three-dimensional intestine-on-chip model was established to emulate these processes in the presence of immune cells, commensal bacteria, and facultative pathogens. Multi-organ tissue flow (MOTiF) biochips made from polystyrene were used for microfluidic perfusion of the intestinal tissue model. The biochips are composed of two chambers separated by a microporous membrane. Each chamber is connected to inlet and outlet channels allowing independent perfusion of the individual channels and application of microfluidic shear stress. Human umbilical vein endothelial cells (HUVECs), monocyte-derived macrophages and intestinal epithelial cells (Caco-2) were assembled on the biochip membrane. Following 7 – 14 days of growth in the presence of physiological flow conditions, the epithelium was colonized with the commensal bacterium Lactobacillus rhamnosus, while the endothelium was perfused with peripheral blood mononuclear cells (PBMCs). Additionally, L. rhamnosus was co-cultivated with the opportunistic fungal pathogen Candida albicans. Within one week of perfusion, the epithelial cells formed self-organized and well-polarized villus- and crypt-like structures that resemble essential morphological characteristics of the human intestine. Dendritic cells were differentiated in the epithelial tissue that specifically responds to bacterial lipopolysaccharide (LPS) challenge. LPS is well-tolerated at the luminal epithelial side of the intestinal model without signs of tissue damage or induction of an inflammatory response, even in the presence of circulating PBMC at the endothelial lining. In contrast, LPS stimulation at the endothelial side of the intestinal model triggered the release of pro-inflammatory cytokines such as TNF, IL-1β, IL-6, and IL-8 via activation of macrophages residing in the endothelium. Perfusion of the endothelium with PBMCs led to an enhanced cytokine release. L. rhamnosus colonization of the model was tolerated in the immune competent tissue model and was demonstrated to reduce damage induced by C. albicans infection. A microfluidic intestine-on-chip model was developed to mimic a systemic infection with a dysregulated immune response under physiological conditions. The model facilitates the colonization of commensal bacteria and co-cultivation with facultative pathogenic microorganisms. Both, commensal bacteria alone and facultative pathogens controlled by commensals, are tolerated by the host and contribute to cell signaling. The human intestine-on-chip model represents a promising tool to mimic microphysiological conditions of the human intestine and paves the way for more detailed in vitro studies of host-microbiota interactions under physiologically relevant conditions.

Keywords: host-microbiota interaction, immune tolerance, microfluidics, organ-on-chip

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56 The Budget Impact of the DISCERN™ Diagnostic Test for Alzheimer’s Disease in the United States

Authors: Frederick Huie, Lauren Fusfeld, William Burchenal, Scott Howell, Alyssa McVey, Thomas F. Goss

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Alzheimer’s Disease (AD) is a degenerative brain disease characterized by memory loss and cognitive decline that presents a substantial economic burden for patients and health insurers in the US. This study evaluates the payer budget impact of the DISCERN™ test in the diagnosis and management of patients with symptoms of dementia evaluated for AD. DISCERN™ comprises three assays that assess critical factors related to AD that regulate memory, formation of synaptic connections among neurons, and levels of amyloid plaques and neurofibrillary tangles in the brain and can provide a quicker, more accurate diagnosis than tests in the current diagnostic pathway (CDP). An Excel-based model with a three-year horizon was developed to assess the budget impact of DISCERN™ compared with CDP in a Medicare Advantage plan with 1M beneficiaries. Model parameters were identified through a literature review and were verified through consultation with clinicians experienced in diagnosis and management of AD. The model assesses direct medical costs/savings for patients based on the following categories: •Diagnosis: costs of diagnosis using DISCERN™ and CDP. •False Negative (FN) diagnosis: incremental cost of care avoidable with a correct AD diagnosis and appropriately directed medication. •True Positive (TP) diagnosis: AD medication costs; cost from a later TP diagnosis with the CDP versus DISCERN™ in the year of diagnosis, and savings from the delay in AD progression due to appropriate AD medication in patients who are correctly diagnosed after a FN diagnosis.•False Positive (FP) diagnosis: cost of AD medication for patients who do not have AD. A one-way sensitivity analysis was conducted to assess the effect of varying key clinical and cost parameters ±10%. An additional scenario analysis was developed to evaluate the impact of individual inputs. In the base scenario, DISCERN™ is estimated to decrease costs by $4.75M over three years, equating to approximately $63.11 saved per test per year for a cohort followed over three years. While the diagnosis cost is higher with DISCERN™ than with CDP modalities, this cost is offset by the higher overall costs associated with CDP due to the longer time needed to receive a TP diagnosis and the larger number of patients who receive a FN diagnosis and progress more rapidly than if they had received appropriate AD medication. The sensitivity analysis shows that the three parameters with the greatest impact on savings are: reduced sensitivity of DISCERN™, improved sensitivity of the CDP, and a reduction in the percentage of disease progression that is avoided with appropriate AD medication. A scenario analysis in which DISCERN™ reduces the utilization for patients of computed tomography from 21% in the base case to 16%, magnetic resonance imaging from 37% to 27% and cerebrospinal fluid biomarker testing, positive emission tomography, electroencephalograms, and polysomnography testing from 4%, 5%, 10%, and 8%, respectively, in the base case to 0%, results in an overall three-year net savings of $14.5M. DISCERN™ improves the rate of accurate, definitive diagnosis of AD earlier in the disease and may generate savings for Medicare Advantage plans.

Keywords: Alzheimer’s disease, budget, dementia, diagnosis.

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55 Prevalence and Associated Risk Factors of Age-Related Macular Degeneration in the Retina Clinic at a Tertiary Center in Makkah Province, Saudi Arabia: A Retrospective Record Review

Authors: Rahaf Mandura, Fatmah Abusharkh, Layan Kurdi, Rahaf Shigdar, Khadijah Alattas

Abstract:

Introduction: Age-related macular degeneration (AMD) in older individuals are serious health issues that severely impact the quality of life of millions globally. In 2020, the fourth leading cause of blindness worldwide was AMD. The global prevalence of AMD is estimated to be around 8.7%. AMD is a progressive disease involving the macular region of the retina, and it has a complex pathophysiology. RPE cell dysfunction plays a crucial step in the pathway leading to irreversible degeneration of photoreceptors with yellowish lipid-rich, protein-containing drusen deposits accumulating between Bruch's membrane and the RPE. Furthermore, lipofuscinogenesis, drusogenesis, inflammation, and neovascularization are four main processes responsible for the formation of the two types of AMD: the wet (exudative, neovascular) and dry (non-exudative, geographic atrophy) types. We retrospectively evaluated the prevalence of AMD among patients visiting the retina clinic at King Abdulaziz University Hospital (Jeddah, Makkah Province, Saudi Arabia) to identify the commonly associated risk factors of AMD. Methods: The records of 3,067 individuals from 2017 to 2021 were reviewed. Of these, 1,935 satisfied the inclusion criteria and were included in this study. We excluded all patient below 18 years, and those who did not undergo fundus imaging or attend their booked appointments, follow-ups, treatments, and referrals were excluded. Results: The prevalence of AMD among the patients was 4%. The age of patients with AMD was significantly greater than those without AMD (72.4 ± 9.8 years vs. 57.2 ± 15.5 years; p < 0.001). Participants with a family history of AMD tended to have the disease more than those without such a history (85.7% vs. 45%; p = 0.043). Ex- and current smokers were more likely to have AMD than non-smokers (34% and 18.6% vs. 7.2%; p < 0.001). Patients with hypertension and those without type 1 diabetes were at a higher risk of developing AMD than those without hypertension (5.5% vs. 2.8%; p = 0.002) and those with type 1 diabetes (4.2% vs. 0.8%; p = 0.040). In contrast, sex, nationality, type 2 diabetes, and abnormal lipid profile were not significantly associated with AMD. Regarding the clinical characteristics of AMD cases, most cases (70.4%) were of the dry type and affected both eyes (77.2%). The disease duration was ≥5 years in 43.1% of the patients. The most frequent chronic diseases associated with AMD were type 2 diabetes (69.1%), hypertension (61.7%), and dyslipidemia (18.5%). Conclusion: In summary, our single tertiary center study showed that AMD is widely prevalent in Jeddah, Saudi Arabia (4%) and linked to a wide range of risk factors. Some of these are modifiable risk factors that can be adjusted to help reduce AMD occurrence. Furthermore, this study has shown the importance of screening and follow-up of family members of patients with AMD to promote early detection and intervention of AMD. We recommend conducting further research on AMD in Saudi Arabia. Concerning the study design, a community-based cross-sectional study would be more helpful for assessing the disease's prevalence. Finally, recruiting a larger sample size is required for more accurate estimation.

Keywords: age related macular degeneration, prevelence, risk factor, dry AMD

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54 Quantification of the Non-Registered Electrical and Electronic Equipment for Domestic Consumption and Enhancing E-Waste Estimation: A Case Study on TVs in Vietnam

Authors: Ha Phuong Tran, Feng Wang, Jo Dewulf, Hai Trung Huynh, Thomas Schaubroeck

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The fast increase and complex components have made waste of electrical and electronic equipment (or e-waste) one of the most problematic waste streams worldwide. Precise information on its size on national, regional and global level has therefore been highlighted as prerequisite to obtain a proper management system. However, this is a very challenging task, especially in developing countries where both formal e-waste management system and necessary statistical data for e-waste estimation, i.e. data on the production, sale and trade of electrical and electronic equipment (EEE), are often lacking. Moreover, there is an inflow of non-registered electronic and electric equipment, which ‘invisibly’ enters the EEE domestic market and then is used for domestic consumption. The non-registration/invisibility and (in most of the case) illicit nature of this flow make it difficult or even impossible to be captured in any statistical system. The e-waste generated from it is thus often uncounted in current e-waste estimation based on statistical market data. Therefore, this study focuses on enhancing e-waste estimation in developing countries and proposing a calculation pathway to quantify the magnitude of the non-registered EEE inflow. An advanced Input-Out Analysis model (i.e. the Sale–Stock–Lifespan model) has been integrated in the calculation procedure. In general, Sale-Stock-Lifespan model assists to improve the quality of input data for modeling (i.e. perform data consolidation to create more accurate lifespan profile, model dynamic lifespan to take into account its changes over time), via which the quality of e-waste estimation can be improved. To demonstrate the above objectives, a case study on televisions (TVs) in Vietnam has been employed. The results show that the amount of waste TVs in Vietnam has increased four times since 2000 till now. This upward trend is expected to continue in the future. In 2035, a total of 9.51 million TVs are predicted to be discarded. Moreover, estimation of non-registered TV inflow shows that it might on average contribute about 15% to the total TVs sold on the Vietnamese market during the whole period of 2002 to 2013. To tackle potential uncertainties associated with estimation models and input data, sensitivity analysis has been applied. The results show that both estimations of waste and non-registered inflow depend on two parameters i.e. number of TVs used in household and the lifespan. Particularly, with a 1% increase in the TV in-use rate, the average market share of non-register inflow in the period 2002-2013 increases 0.95%. However, it decreases from 27% to 15% when the constant unadjusted lifespan is replaced by the dynamic adjusted lifespan. The effect of these two parameters on the amount of waste TV generation for each year is more complex and non-linear over time. To conclude, despite of remaining uncertainty, this study is the first attempt to apply the Sale-Stock-Lifespan model to improve the e-waste estimation in developing countries and to quantify the non-registered EEE inflow to domestic consumption. It therefore can be further improved in future with more knowledge and data.

Keywords: e-waste, non-registered electrical and electronic equipment, TVs, Vietnam

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53 The Concept of Path in Original Buddhism and the Concept of Psychotherapeutic Improvement

Authors: Beth Jacobs

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The landmark movement of Western clinical psychology in the 20th century was the development of psychotherapy. The landmark movement of clinical psychology in the 21st century will be the absorption of meditation practices from Buddhist psychology. While millions of people explore meditation and related philosophy, very few people are exposed to the materials of original Buddhism on this topic, especially to the Theravadan Abhidharma. The Abhidharma is an intricate system of lists and matrixes that were used to understand and remember Buddha’s teaching. The Abhidharma delineates the first psychological system of Buddhism, how the mind works in the universe of reality and why meditation training strengthens and purifies the experience of life. Its lists outline the psychology of mental constructions, perception, emotion and cosmological causation. While the Abhidharma is technical, elaborate and complex, its essential purpose relates to the central purpose of clinical psychology: to relieve human suffering. Like Western depth psychology, the methodology rests on understanding underlying processes of consciousness and perception. What clinical psychologists might describe as therapeutic improvement, the Abhidharma delineates as a specific pathway of purified actions of consciousness. This paper discusses the concept of 'path' as presented in aspects of the Theravadan Abhidharma and relates this to current clinical psychological views of therapy outcomes and gains. The core path in Buddhism is the Eight-Fold Path, which is the fourth noble truth and the launching of activity toward liberation. The path is not composed of eight ordinal steps; it’s eight-fold and is described as opening the way, not funneling choices. The specific path in the Abhidharma is described in many steps of development of consciousness activities. The path is not something a human moves on, but something that moments of consciousness develop within. 'Cittas' are extensively described in the Abhidharma as the atomic-level unit of a raw action of consciousness touching upon an object in a field, and there are 121 types of cittas categorized. The cittas are embedded in the mental factors, which could be described as the psychological packaging elements of our experiences of consciousness. Based on these constellations of infinitesimal, linked occurrences of consciousness, citta are categorized by dimensions of purification. A path is a chain of citta developing through causes and conditions. There are no selves, no pronouns in the Abhidharma. Instead of me walking a path, this is about a person working with conditions to cultivate a stream of consciousness that is pure, immediate, direct and generous. The same effort, in very different terms, informs the work of most psychotherapies. Depth psychology seeks to release the bound, unconscious elements of mental process into the clarity of realization. Cognitive and behavioral psychologies work on breaking down automatic thought valuations and actions, changing schemas and interpersonal dynamics. Understanding how the original Buddhist concept of positive human development relates to the clinical psychological concept of therapy weaves together two brilliant systems of thought on the development of human well being.

Keywords: Abhidharma, Buddhist path, clinical psychology, psychotherapeutic outcome

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52 Exploring the Impact of Mobility-Related Treatments (Drug and Non-Pharmacological) on Independence and Wellbeing in Parkinson’s Disease - A Qualitative Synthesis

Authors: Cameron Wilson, Megan Hanrahan, Katie Brittain, Riona McArdle, Alison Keogh, Lynn Rochester

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Background: The loss of mobility and functional dependence is a significant marker in the progression of neurodegenerative diseases such as Parkinson’s Disease (PD). Pharmacological, surgical, and therapeutic treatments are available that can help in the management and amelioration of PD symptoms; however, these only prolong more severe symptoms. Accordingly, ensuring people with PD can maintain independence and a healthy wellbeing are essential in establishing an effective treatment option for those afflicted. Existing literature reviews have examined experiences in engaging with PD treatment options and the impact of PD on independence and wellbeing. Although, the literature fails to explore the influence of treatment options on independence and wellbeing and therefore misses what people value in their treatment. This review is the first that synthesises the impact of mobility-related treatments on independence and wellbeing in people with PD and their carers, offering recommendations to clinical practice and provides a conceptual framework (in development) for future research and practice. Objectives: To explore the impact of mobility-related treatment (both pharmacological and non-pharmacological) on the independence and wellbeing of people with PD and their carers. To propose a conceptual framework to patients, carers and clinicians which captures the qualities people with PD value as part of their treatment. Methods: We performed a critical interpretive synthesis of qualitative evidence, searching six databases for reports that explored the impact of mobility-related treatments (both drug and non-pharmacological) on independence and wellbeing in Parkinson’s Disease. The types of treatments included medication (Levodopa and Amantadine), dance classes, Deep-Brain Stimulation, aquatic therapies, physical rehabilitation, balance training and foetal transplantation. Data was extracted, and quality was assessed using an adapted version of the NICE Quality Appraisal Tool Appendix H before being synthesised according to the critical interpretive synthesis framework and meta-ethnography process. Results: From 2301 records, 28 were eligible. Experiences and impact of treatment pathway on independence and wellbeing was similar across all types of treatments and are described by five inter-related themes: (i) desire to maintain independence, (ii) treatment as a social experience during and after, (iii) medication to strengthen emotional health, (iv) recognising physical capacity and (v) emphasising the personal journey of Parkinson’s treatments. Conclusion: There is a complex and inter-related experience and effect of PD treatments common across all types of treatment. The proposed conceptual framework (in development) provides patients, carers, and clinicians recommendations to personalise the delivery of PD treatment, thereby potentially improving adherence and effectiveness. This work is vital to disseminate as PD treatment transitions from subjective and clinically captured assessments to a more personalised process supplemented using wearable technology.

Keywords: parkinson's disease, medication, treatment, dance, review, healthcare, delivery, levodopa, social, emotional, psychological, personalised healthcare

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51 Impact of Elevated Temperature on Spot Blotch Development in Wheat and Induction of Resistance by Plant Growth Promoting Rhizobacteria

Authors: Jayanwita Sarkar, Usha Chakraborty, Bishwanath Chakraborty

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Plants are constantly interacting with various abiotic and biotic stresses. In changing climate scenario plants are continuously modifying physiological processes to adapt to changing environmental conditions which profoundly affect plant-pathogen interactions. Spot blotch in wheat is a fast-rising disease in the warmer plains of South Asia where the rise in minimum average temperature over most of the year already affecting wheat production. Hence, the study was undertaken to explore the role of elevated temperature in spot blotch disease development and modulation of antioxidative responses by plant growth promoting rhizobacteria (PGPR) for biocontrol of spot blotch at high temperature. Elevated temperature significantly increases the susceptibility of wheat plants to spot blotch causing pathogen Bipolaris sorokiniana. Two PGPR Bacillus safensis (W10) and Ochrobactrum pseudogrignonense (IP8) isolated from wheat (Triticum aestivum L.) and blady grass (Imperata cylindrical L.) rhizophere respectively, showing in vitro antagonistic activity against Bipolaris sorokiniana were tested for growth promotion and induction of resistance against spot blotch in wheat. GC-MS analysis showed that Bacillus safensis (W10) and Ochrobactrum pseudogrignonense (IP8) produced antifungal and antimicrobial compounds in culture. Seed priming with these two bacteria significantly increase growth, modulate antioxidative signaling and induce resistance and eventually reduce disease incidence in wheat plants at optimum as well as elevated temperature which was further confirmed by indirect immunofluorescence assay using polyclonal antibody raised against Bipolaris sorokiniana. Application of the PGPR led to enhancement in activities of plant defense enzymes- phenylalanine ammonia lyase, peroxidase, chitinase and β-1,3 glucanase in infected leaves. Immunolocalization of chitinase and β-1,3 glucanase in PGPR primed and pathogen inoculated leaf tissue was further confirmed by transmission electron microscopy using PAb of chitinase, β-1,3 glucanase and gold labelled conjugates. Activity of ascorbate-glutathione redox cycle related enzymes such as ascorbate peroxidase, superoxide dismutase and glutathione reductase along with antioxidants such as carotenoids, glutathione and ascorbate and osmolytes like proline and glycine betain accumulation were also increased during disease development in PGPR primed plant in comparison to unprimed plants at high temperature. Real-time PCR analysis revealed enhanced expression of defense genes- chalcone synthase and phenyl alanineammonia lyase. Over expression of heat shock proteins like HSP 70, small HSP 26.3 and heat shock factor HsfA3 in PGPR primed plants effectively protect plants against spot blotch infection at elevated temperature as compared with control plants. Our results revealed dynamic biochemical cross talk between elevated temperature and spot blotch disease development and furthermore highlight PGPR mediated array of antioxidative and molecular alterations responsible for induction of resistance against spot blotch disease at elevated temperature which seems to be associated with up-regulation of defense genes, heat shock proteins and heat shock factors, less ROS production, membrane damage, increased expression of redox enzymes and accumulation of osmolytes and antioxidants.

Keywords: antioxidative enzymes, defense enzymes, elevated temperature, heat shock proteins, PGPR, Real-Time PCR, spot blotch, wheat

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50 Effects of Oxidized LDL in M2 Macrophages: Implications in Atherosclerosis

Authors: Fernanda Gonçalves, Karla Alcântara, Vanessa Moura, Patrícia Nolasco, Jorge Kalil, Maristela Hernandez

Abstract:

Introduction: Atherosclerosis is a chronic disease where two striking features are observed: retention of lipids and inflammation. Understanding the interaction between immune cells and lipoproteins involved in atherogenesis are urgent challenges, since cardiovascular diseases are the leading cause of death worldwide. Macrophages are critical to the development of atherosclerotic plaques and in the perpetuation of inflammation in these lesions. These cells are also directly involved in unstable plaque rupture. Recently different populations of macrophages are being identified in atherosclerotic lesions. Although the presence of M2 macrophages (macrophages activated by the alternative pathway, eg. The IL-4) has been identified, the function of these cells in atherosclerosis is not yet defined. M2 macrophages have a high endocytic capacity, they promote remodeling of tissues and to have anti-inflammatory activity. However, in atherosclerosis, especially unstable plaques, severe inflammatory reaction, accumulation of cellular debris and intense degradation of the tissue is observed. Thus, it is possible that the M2 macrophages have altered function (phenotype) in atherosclerosis. Objective: Our aim is to evaluate if the presence of oxidized LDL alters the phenotype and function of M2 macrophages in vitro. Methods: For this, we will evaluate whether the addition of lipoprotein in M2 macrophages differentiated in vitro with IL -4 induces 1) a reduction in the secretion of anti-inflammatory cytokines (CBA and ELISA), 2) secretion of inflammatory cytokines (CBA and ELISA), 3) expression of cell activation markers (Flow cytometry), 4) alteration in gene expression of molecules adhesion and extracellular matrix (Real-Time PCR) and 5) Matrix degradation (confocal microscopy). Results: In oxLDL stimulated M2 macrophages cultures we did not find any differences in the expression of the cell surface markers tested, including: HLA-DR, CD80, CD86, CD206, CD163 and CD36. Also, cultures stimulated with oxLDL had similar phagocytic capacity when compared to unstimulated cells. However, in the supernatant of these cultures an increase in the secretion of the pro-inflammatory cytokine IL-8 was detected. No significant changes where observed in IL-6, IL-10, IL-12 and IL-1b levels. The culture supernatant also induced massive extracellular matrix (produced by mouse embryo fibroblast) filaments degradation. When evaluating the expression of 84 extracellular matrix and adhesion molecules genes, we observed that the stimulation of oxLDL in M2 macrophages decreased 47% of the genes and increased the expression of only 3% of the genes. In particular we noted that oxLDL inhibit the expression of 60% of the genes constituents of extracellular matrix and collagen expressed by these cells, including fibronectin1 and collagen VI. We also observed a decrease in the expression of matrix protease inhibitors, such as TIMP 2. On the opposite, the matricellular protein thrombospondin had a 12 fold increase in gene expression. In the presence of native LDL 90% of the genes had no altered expression. Conclusion: M2 macrophages stimulated with oxLDL secrete the pro-inflammatory cytokine IL-8, have an altered extracellular matrix constituents gene expression, and promote the degradation of extracellular matrix. M2 macrophages may contribute to the perpetuation of inflammation in atherosclerosis and to plaque rupture.

Keywords: atherosclerosis, LDL, macrophages, m2

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49 Molecular Signaling Involved in the 'Benzo(a)Pyrene' Induced Germ Cell DNA Damage and Apoptosis: Possible Protection by Natural Aryl Hydrocarbon Receptor Antagonist and Anti-Tumor Agent

Authors: Kuladip Jana

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Benzo(a)pyrene [B(a)P] is an environmental toxicant present mostly in cigarette smoke and car exhaust, is an aryl hydrocarbon receptor (AhR) ligand that exerts its toxic effects on both male and female reproductive systems. In this study, the effect of B(a)P at different doses (0.1, 0.25, 0.5, 1 and 5 mg /kg body weight) was studied on male reproductive system of rat. A significant decrease in cauda epididymal sperm count and motility along with the presence of sperm head abnormalities and altered epididymal and testicular histology were documented following B(a)P treatment. B(a)P treatment resulted apoptotic sperm cells as observed by TUNEL and Annexin V-PI assay with increased ROS, altered sperm mitochondrial membrane potential (ΔΨm) with a simultaneous decrease in the activity of antioxidant enzymes and GSH status. TUNEL positive apoptotic cells also observed in testis as well as isolated germ and Leydig cells following B(a)P exposure. Western Blot analysis revealed the activation of p38MAPK, cytosolic translocation of cytochrome-c, up-regulation of Bax and inducible nitric oxide synthase (iNOS) with cleavage of PARP and down-regulation of BCl2 in testis upon B(a)P treatment. The protein and mRNA levels of testicular key steroidogenesis regulatory proteins like StAR, cytochrome P450 IIA1 (CYPIIA1), 3β HSD, 17β HSD showed a significant decrease in a dose dependent manner while an increase in the expression of cytochrome P450 1A1 (CYP1A1), Aryl hydrocarbon Receptor (AhR), active caspase- 9 and caspase- 3 following B(a)P exposure. We conclude that exposure of benzo(a)pyrene caused testicular gamatogenic and steroidogenic disorders by induction of oxidative stress, inhibition of StAR and other steroidogenic enzymes along with activation of p38MAPK and initiated caspase-3 mediated germ and Leydig cell apoptosis.The possible protective role of naturally occurring phytochemicals against B(a)P induced testicular toxicity needs immediate consideration. Curcumin and resveratrol separately were found to protect against B(a)P induced germ cell apoptosis, and their combinatorial effect was more significant. Our present study in isolated testicular germ cell population from adult male Wistar rats, highlighted their synergistic protective effect against B(a)P induced germ cell apoptosis. Curcumin-resveratrol co-treatment decreased the expression of pro-apoptotic proteins like cleaved caspase 3,8,9, cleaved PARP, Apaf1, FasL, tBid. Curcumin-resveratrol co-treatment decreased Bax/Bcl2 ratio, mitochondria to cytosolic translocation of cytochrome c and activated the survival protein Akt. Curcumin-resveratrol decreased the expression of p53 dependent apoptotic genes like Fas, FasL, Bax, Bcl2, Apaf1.Curcumin-resveratrol co-treatment thus prevented B(a)P induced germ cell apoptosis. B(a)P induced testicular ROS generation and oxidative stress were significantly ameliorated with curcumin and resveratrol. Curcumin-resveratrol co-treatment prevented B(a)P induced nuclear translocation of AhR and CYP1A1 production. The combinatorial treatment significantly inhibited B(a)P induced ERK 1/2, p38 MAPK and JNK 1/2 activation. B(a)P treatment increased the expression of p53 and its phosphorylation (p53 ser 15). Curcumin-resveratrol co-treatment significantly decreased p53 level and its phosphorylation (p53 ser 15). The study concludes that curcumin-resveratrol synergistically modulated MAPKs and p53, prevented oxidative stress, regulated the expression of pro and anti-apoptotic proteins as well as the proteins involved in B(a)P metabolism thus protected germ cells from B(a)P induced apoptosis.

Keywords: benzo(a)pyrene, germ cell, apoptosis, oxidative stress, resveratrol, curcumin

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48 Engineering Economic Analysis of Implementing a Materials Recovery Facility in Jamaica: A Green Industry Approach towards a Sustainable Developing Economy

Authors: Damian Graham, Ashleigh H. Hall, Damani R. Sulph, Michael A. James, Shawn B. Vassell

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This paper assesses the design and feasibility of a Materials Recovery Facility (MRF) in Jamaica as a possible green industry approach to the nation’s economic and solid waste management problems. Jamaica is a developing nation that is vulnerable to climate change that can affect its blue economy and tourism on which it is heavily reliant. Jamaica’s National Solid Waste Management Authority (NSWMA) collects only a fraction of all the solid waste produced annually which is then transported to dumpsites. The remainder is either burnt by the population or disposed of illegally. These practices negatively impact the environment, threaten the sustainability of economic growth from blue economy and tourism and its waste management system is predominantly a cost centre. The implementation of an MRF could boost the manufacturing sector, contribute to economic growth, and be a catalyst in creating a green industry with multiple downstream value chains with supply chain linkages. Globally, there is a trend to reuse and recycle that created an international market for recycled solid waste. MRFs enable the efficient sorting of solid waste into desired recoverable materials thus providing a gateway for entrance to the international trading of recycled waste. Research into the current state and effort to improve waste management in Jamaica in contrast with the similar and more advanced territories are outlined. The study explores the concept of green industrialization and its applicability to vulnerable small state economies like Jamaica. The study highlights the possible contributions and benefits derived from MRFs as a seeding factory that can anchor the reverse and forward logistics of other green industries as part of a logistic-cantered economy. Further, the study showcases an engineering economic analysis that assesses the viability of the implementation of an MRF in Jamaica. This research outlines the potential cost of constructing and operating an MRF and provides a realistic cash flow estimate to establish a baseline for profitability. The approach considers quantitative and qualitative data, assumptions, and modelling using industrial engineering tools and techniques that are outlined. Techniques of facility planning, system analysis and operations research with a focus on linear programming techniques are expressed. Approaches to overcome some implementation challenges including policy, technology and public education are detailed. The results of this study present a reasonable judgment of the prospects of incorporating an MRF to improve Jamaica’s solid waste management and contribute to socioeconomic and environmental benefits and an alternate pathway for economic sustainability.

Keywords: engineering-economic analysis, facility design, green industry, MRF, manufacturing, plant layout, solid-waste management, sustainability, waste disposal

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47 Analysis of Taxonomic Compositions, Metabolic Pathways and Antibiotic Resistance Genes in Fish Gut Microbiome by Shotgun Metagenomics

Authors: Anuj Tyagi, Balwinder Singh, Naveen Kumar B. T., Niraj K. Singh

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Characterization of diverse microbial communities in specific environment plays a crucial role in the better understanding of their functional relationship with the ecosystem. It is now well established that gut microbiome of fish is not the simple replication of microbiota of surrounding local habitat, and extensive species, dietary, physiological and metabolic variations in fishes may have a significant impact on its composition. Moreover, overuse of antibiotics in human, veterinary and aquaculture medicine has led to rapid emergence and propagation of antibiotic resistance genes (ARGs) in the aquatic environment. Microbial communities harboring specific ARGs not only get a preferential edge during selective antibiotic exposure but also possess the significant risk of ARGs transfer to other non-resistance bacteria within the confined environments. This phenomenon may lead to the emergence of habitat-specific microbial resistomes and subsequent emergence of virulent antibiotic-resistant pathogens with severe fish and consumer health consequences. In this study, gut microbiota of freshwater carp (Labeo rohita) was investigated by shotgun metagenomics to understand its taxonomic composition and functional capabilities. Metagenomic DNA, extracted from the fish gut, was subjected to sequencing on Illumina NextSeq to generate paired-end (PE) 2 x 150 bp sequencing reads. After the QC of raw sequencing data by Trimmomatic, taxonomic analysis by Kraken2 taxonomic sequence classification system revealed the presence of 36 phyla, 326 families and 985 genera in the fish gut microbiome. At phylum level, Proteobacteria accounted for more than three-fourths of total bacterial populations followed by Actinobacteria (14%) and Cyanobacteria (3%). Commonly used probiotic bacteria (Bacillus, Lactobacillus, Streptococcus, and Lactococcus) were found to be very less prevalent in fish gut. After sequencing data assembly by MEGAHIT v1.1.2 assembler and PROKKA automated analysis pipeline, pathway analysis revealed the presence of 1,608 Metacyc pathways in the fish gut microbiome. Biosynthesis pathways were found to be the most dominant (51%) followed by degradation (39%), energy-metabolism (4%) and fermentation (2%). Almost one-third (33%) of biosynthesis pathways were involved in the synthesis of secondary metabolites. Metabolic pathways for the biosynthesis of 35 antibiotic types were also present, and these accounted for 5% of overall metabolic pathways in the fish gut microbiome. Fifty-one different types of antibiotic resistance genes (ARGs) belonging to 15 antimicrobial resistance (AMR) gene families and conferring resistance against 24 antibiotic types were detected in fish gut. More than 90% ARGs in fish gut microbiome were against beta-lactams (penicillins, cephalosporins, penems, and monobactams). Resistance against tetracycline, macrolides, fluoroquinolones, and phenicols ranged from 0.7% to 1.3%. Some of the ARGs for multi-drug resistance were also found to be located on sequences of plasmid origin. The presence of pathogenic bacteria and ARGs on plasmid sequences suggested the potential risk due to horizontal gene transfer in the confined gut environment.

Keywords: antibiotic resistance, fish gut, metabolic pathways, microbial diversity

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46 Play, Practice and Perform: The Pathway to Becoming and Belonging as an Engineer

Authors: Rick Evans

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Despite over 40 years of research into why women choose not to enroll or leave undergraduate engineering programs, along with the subsequent and serious efforts to attract more women, women receiving bachelor's degrees in engineering in the US have remained disappointingly low. We know that even despite their struggles to become more welcoming and inclusive, engineering programs remain gendered, raced and classed. However, our research team has found that women who participate and indeed thrive in undergraduate engineering project teams do so in numbers that far exceed their participation in undergraduate programs. We believe part of the answer lies in the ways that project teams facilitate experiential learning, specifically providing opportunities for members to play, practice and perform. We employ a multi-case study method and assume a feminist, activist and interpretive perspective. We seek to generate concrete and context-dependent knowledge in order to explore potentially new variables and hypotheses. Our focus is to learn from those select women who are thriving. For this oral or e-poster presentation, we will focus on the results of the second of our semi-structured interviews – the learning journey interview. During this interview, we ask participants to tell us the story/ies of their participation in project teams. Our results suggest these women find joy in their experience of developing and applying engineering expertise. They experience this joy and develop their expertise in the highly patterned progression of play, practice and performance. Play is a purposeful activity in which someone enters an imaginary world, a world not yet real to them. However, this imaginary world is still very much connected to the real world, in this case, a particular kind of engineering, in that the ways of engaging are already established, codified and rule-governed. As such, these women are novices motivated to join a community of actors. Practice, better understood as practices, a count noun, is an embodied, materially interconnected collection of actions organized around the shared understandings of that community of actors. Those shared understandings reveal a social order – a particular field of engineering. No longer novices, these women begin to develop and display their emergent identities as engineers. Perform is activity meant either to demonstrate competence and/or to enable, even teach play and practice to others. As performers, these women participants become models for others. They direct play and practice, contextualizing both within a field of engineering and the specific aims of the project team community. By playing, practicing and performing engineering, women claim their identities as engineers and, equally important, have those identities acknowledged by team members. If we hope to transform our gendered, raced, classed institutions, we need to learn more about women who thrive within those institutions. We need to learn more about their processes of becoming and belonging as engineers. Our research presentation begins with a description of project teams and our multi-case study method. We then offer detailed descriptions of play, practice, and performance using the voices of women in project teams.

Keywords: engineering education, gender, identity, project teams

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45 Isoflavonoid Dynamic Variation in Red Clover Genotypes

Authors: Andrés Quiroz, Emilio Hormazábal, Ana Mutis, Fernando Ortega, Loreto Méndez, Leonardo Parra

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Red clover root borer, Hylastinus obscurus Marsham (Coleoptera: Curculionidae), is the main insect pest associated to red clover, Trifolium pratense L. An average of 1.5 H. obscurus per plant can cause 5.5% reduction in forage yield in pastures of two to three years old. Moreover, insect attack can reach 70% to 100% of the plants. To our knowledge, there is no a chemical strategy for controlling this pest. Therefore alternative strategies for controlling H. obscurus are a high priority for red clover producers. One of this alternative is related to the study of secondary metabolites involved in intrinsic chemical defenses developed by plants, such as isoflavonoids. The isoflavonoids formononetin and daidzein have elicited an antifeedant and phagostimult effect on H. obscurus respectively. However, we do not know how is the dynamic variation of these isoflavonoids under field conditions. The main objective of this work was to evaluate the variation of the antifeedant isoflavonoids formononetin, the phagostimulant isoflavonoids daidzein, and their respective glycosides over time in different ecotypes of red clover. Fourteen red clover ecotypes (8 cultivars and 6 experimental lines), were collected at INIA-Carillanca (La Araucanía, Chile). These plants were established in October 2015 under irrigated conditions. The cultivars were distributed in a randomized complete block with three replicates. The whole plants were sampled in four times: 15th October 2016, 12th December 2016, 27th January 2017 and 16th March 2017 with sufficient amount of soil to avoid root damage. A polar fraction of isoflavonoid was obtained from 20 mg of lyophilized root tissue extracted with 2 mL of 80% MeOH for 16 h using an orbital shaker in the dark at room temperature. After, an aliquot of 1.4 mL of the supernatant was evaporated, and the residue was resuspended in 300 µL of 45% MeOH. The identification and quantification of isoflavonoid root extracts were performed by the injection of 20 µL into a Shimadzu HPLC equipped with a C-18 column. The sample was eluted with a mobile phase composed of AcOH: H₂O (1:9 v/v) as solvent A and CH₃CN as solvent B. The detection was performed at 260 nm. The results showed that the amount of aglycones was higher than the respective glycosides. This result is according to the biosynthetic pathway of flavonoids, where the formation of glycoside is further to the glycosides biosynthesis. The amount of formononetin was higher than daidzein. In roots, where H. obscurus spent the most part of its live cycle, the highest content of formononetin was found in G 27, Pawera, Sabtoron High, Redqueli-INIA and Superqueli-INIA cvs. (2.1, 1.8, 1.8, 1.6 and 1.0 mg g⁻¹ respectively); and the lowest amount of daidzein were found Superqueli-INIA (0.32 mg g⁻¹) and in the experimental line Sel Syn Int4 (0.24 mg g⁻¹). This ecotype showed a high content of formononetin (0.9 mg g⁻¹). This information, associated with cultural practices, could help farmers and breeders to reduce H. obscurus in grassland, selecting ecotypes with high content of formononetin and low amount of daidzein in the roots of red clover plants. Acknowledgements: FONDECYT 1141245 and 11130715.

Keywords: daidzein, formononetin, isoflavonoid glycosides, trifolium pratense

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