Search results for: combining ability
Commenced in January 2007
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Paper Count: 5041

Search results for: combining ability

211 Identification and Characterization of Small Peptides Encoded by Small Open Reading Frames using Mass Spectrometry and Bioinformatics

Authors: Su Mon Saw, Joe Rothnagel

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Short open reading frames (sORFs) located in 5’UTR of mRNAs are known as uORFs. Characterization of uORF-encoded peptides (uPEPs) i.e., a subset of short open reading frame encoded peptides (sPEPs) and their translation regulation lead to understanding of causes of genetic disease, proteome complexity and development of treatments. Existence of uORFs within cellular proteome could be detected by LC-MS/MS. The ability of uORF to be translated into uPEP and achievement of uPEP identification will allow uPEP’s characterization, structures, functions, subcellular localization, evolutionary maintenance (conservation in human and other species) and abundance in cells. It is hypothesized that a subset of sORFs are translatable and that their encoded sPEPs are functional and are endogenously expressed contributing to the eukaryotic cellular proteome complexity. This project aimed to investigate whether sORFs encode functional peptides. Liquid chromatography-mass spectrometry (LC-MS) and bioinformatics were thus employed. Due to probable low abundance of sPEPs and small in sizes, the need for efficient peptide enrichment strategies for enriching small proteins and depleting the sub-proteome of large and abundant proteins is crucial for identifying sPEPs. Low molecular weight proteins were extracted using SDS-PAGE from Human Embryonic Kidney (HEK293) cells and Strong Cation Exchange Chromatography (SCX) from secreted HEK293 cells. Extracted proteins were digested by trypsin to peptides, which were detected by LC-MS/MS. The MS/MS data obtained was searched against Swiss-Prot using MASCOT version 2.4 to filter out known proteins, and all unmatched spectra were re-searched against human RefSeq database. ProteinPilot v5.0.1 was used to identify sPEPs by searching against human RefSeq, Vanderperre and Human Alternative Open Reading Frame (HaltORF) databases. Potential sPEPs were analyzed by bioinformatics. Since SDS PAGE electrophoresis could not separate proteins <20kDa, this could not identify sPEPs. All MASCOT-identified peptide fragments were parts of main open reading frame (mORF) by ORF Finder search and blastp search. No sPEP was detected and existence of sPEPs could not be identified in this study. 13 translated sORFs in HEK293 cells by mass spectrometry in previous studies were characterized by bioinformatics. Identified sPEPs from previous studies were <100 amino acids and <15 kDa. Bioinformatics results showed that sORFs are translated to sPEPs and contribute to proteome complexity. uPEP translated from uORF of SLC35A4 was strongly conserved in human and mouse while uPEP translated from uORF of MKKS was strongly conserved in human and Rhesus monkey. Cross-species conserved uORFs in association with protein translation strongly suggest evolutionary maintenance of coding sequence and indicate probable functional expression of peptides encoded within these uORFs. Translation of sORFs was confirmed by mass spectrometry and sPEPs were characterized with bioinformatics.

Keywords: bioinformatics, HEK293 cells, liquid chromatography-mass spectrometry, ProteinPilot, Strong Cation Exchange Chromatography, SDS-PAGE, sPEPs

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210 Supporting a Moral Growth Mindset Among College Students

Authors: Kate Allman, Heather Maranges, Elise Dykhuis

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Moral Growth Mindset (MGM) is the belief that one has the capacity to become a more moral person, as opposed to a fixed conception of one’s moral ability and capacity (Han et al., 2018). Building from Dweck’s work in incremental implicit theories of intelligence (2008), Moral Growth Mindset (Han et al., 2020) extends growth mindsets into the moral dimension. The concept of MGM has the potential to help researchers understand how both mindsets and interventions can impact character development, and it has even been shown to have connections to voluntary service engagement (Han et al., 2018). Understanding the contexts in which MGM might be cultivated could help to promote the further cultivation of character, in addition to prosocial behaviors like service engagement, which may, in turn, promote larger scale engagement in social justice-oriented thoughts, feelings, and behaviors. In particular, college may be a place to intentionally cultivate a growth mindset toward moral capacities, given the unique developmental and maturational components of the college experience, including contextual opportunity (Lapsley & Narvaez, 2006) and independence requiring the constant consideration, revision, and internalization of personal values (Lapsley & Woodbury, 2016). In a semester-long, quasi-experimental study, we examined the impact of a pedagogical approach designed to cultivate college student character development on participants’ MGM. With an intervention (n=69) and a control group (n=97; Pre-course: 27% Men; 66% Women; 68% White; 18% Asian; 2% Black; <1% Hispanic/Latino), we investigated whether college courses that intentionally incorporate character education pedagogy (Lamb, Brant, Brooks, 2021) affect a variety of psychosocial variables associated with moral thoughts, feelings, identity, and behavior (e.g. moral growth mindset, honesty, compassion, etc.). The intervention group consisted of 69 undergraduate students (Pre-course: 40% Men; 52% Women; 68% White; 10.5% Black; 7.4% Asian; 4.2% Hispanic/Latino) that voluntarily enrolled in five undergraduate courses that encouraged students to engage with key concepts and methods of character development through the application of research-based strategies and personal reflection on goals and experiences. Moral Growth Mindset was measured using the four-item Moral Growth Mindset scale (Han et al., 2020), with items such as You can improve your basic morals and character considerably on a six-point Likert scale from 1 (strongly disagree) to 6 (strongly agree). Higher scores of MGM indicate a stronger belief that one can become a more moral person with personal effort. Reliability at Time 1 was Cronbach’s ɑ= .833, and at Time 2 Cronbach’s ɑ= .772. An Analysis of Covariance (ANCOVA) was conducted to explore whether post-course MGM scores were different between the intervention and control when controlling for pre-course MGM scores. The ANCOVA indicated significant differences in MGM between groups post-course, F(1,163) = 8.073, p = .005, R² = .11, where descriptive statistics indicate that intervention scores were higher than the control group at post-course. Results indicate that intentional character development pedagogy can be leveraged to support the development of Moral Growth Mindset and related capacities in undergraduate settings.

Keywords: moral personality, character education, incremental theories of personality, growth mindset

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209 Effect of Non-Thermal Plasma, Chitosan and Polymyxin B on Quorum Sensing Activity and Biofilm of Pseudomonas aeruginosa

Authors: Alena Cejkova, Martina Paldrychova, Jana Michailidu, Olga Matatkova, Jan Masak

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Increasing the resistance of pathogenic microorganisms to many antibiotics is a serious threat to the treatment of infectious diseases and cleaning medical instruments. It should be added that the resistance of microbial populations growing in biofilms is often up to 1000 times higher compared to planktonic cells. Biofilm formation in a number of microorganisms is largely influenced by the quorum sensing regulatory mechanism. Finding external factors such as natural substances or physical processes that can interfere effectively with quorum sensing signal molecules should reduce the ability of the cell population to form biofilm and increase the effectiveness of antibiotics. The present work is devoted to the effect of chitosan as a representative of natural substances with anti-biofilm activity and non- thermal plasma (NTP) alone or in combination with polymyxin B on biofilm formation of Pseudomonas aeruginosa. Particular attention was paid to the influence of these agents on the level of quorum sensing signal molecules (acyl-homoserine lactones) during planktonic and biofilm cultivations. Opportunistic pathogenic strains of Pseudomonas aeruginosa (DBM 3081, DBM 3777, ATCC 10145, ATCC 15442) were used as model microorganisms. Cultivations of planktonic and biofilm populations in 96-well microtiter plates on horizontal shaker were used for determination of antibiotic and anti-biofilm activity of chitosan and polymyxin B. Biofilm-growing cells on titanium alloy, which is used for preparation of joint replacement, were exposed to non-thermal plasma generated by cometary corona with a metallic grid for 15 and 30 minutes. Cultivation followed in fresh LB medium with or without chitosan or polymyxin B for next 24 h. Biofilms were quantified by crystal violet assay. Metabolic activity of the cells in biofilm was measured using MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) colorimetric test based on the reduction of MTT into formazan by the dehydrogenase system of living cells. Activity of N-acyl homoserine lactones (AHLs) compounds involved in the regulation of biofilm formation was determined using Agrobacterium tumefaciens strain harboring a traG::lacZ/traR reporter gene responsive to AHLs. The experiments showed that both chitosan and non-thermal plasma reduce the AHLs level and thus the biofilm formation and stability. The effectiveness of both agents was somewhat strain dependent. During the eradication of P. aeruginosa DBM 3081 biofilm on titanium alloy induced by chitosan (45 mg / l) there was an 80% decrease in AHLs. Applying chitosan or NTP on the P. aeruginosa DBM 3777 biofilm did not cause a significant decrease in AHLs, however, in combination with both (chitosan 55 mg / l and NTP 30 min), resulted in a 70% decrease in AHLs. Combined application of NTP and polymyxin B allowed reduce antibiotic concentration to achieve the same level of AHLs inhibition in P. aeruginosa ATCC 15442. The results shown that non-thermal plasma and chitosan have considerable potential for the eradication of highly resistant P. aeruginosa biofilms, for example on medical instruments or joint implants.

Keywords: anti-biofilm activity, chitosan, non-thermal plasma, opportunistic pathogens

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208 Liposome Loaded Polysaccharide Based Hydrogels: Promising Delayed Release Biomaterials

Authors: J. Desbrieres, M. Popa, C. Peptu, S. Bacaita

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Because of their favorable properties (non-toxicity, biodegradability, mucoadhesivity etc.), polysaccharides were studied as biomaterials and as pharmaceutical excipients in drug formulations. These formulations may be produced in a wide variety of forms including hydrogels, hydrogel based particles (or capsules), films etc. In these formulations, the polysaccharide based materials are able to provide local delivery of loaded therapeutic agents but their delivery can be rapid and not easily time-controllable due to, particularly, the burst effect. This leads to a loss in drug efficiency and lifetime. To overcome the consequences of burst effect, systems involving liposomes incorporated into polysaccharide hydrogels may appear as a promising material in tissue engineering, regenerative medicine and drug loading systems. Liposomes are spherical self-closed structures, composed of curved lipid bilayers, which enclose part of the surrounding solvent into their structure. The simplicity of production, their biocompatibility, the size and similar composition of cells, the possibility of size adjustment for specific applications, the ability of hydrophilic or/and hydrophobic drug loading make them a revolutionary tool in nanomedicine and biomedical domain. Drug delivery systems were developed as hydrogels containing chitosan or carboxymethylcellulose (CMC) as polysaccharides and gelatin (GEL) as polypeptide, and phosphatidylcholine or phosphatidylcholine/cholesterol liposomes able to accurately control this delivery, without any burst effect. Hydrogels based on CMC were covalently crosslinked using glutaraldehyde, whereas chitosan based hydrogels were double crosslinked (ionically using sodium tripolyphosphate or sodium sulphate and covalently using glutaraldehyde). It has been proven that the liposome integrity is highly protected during the crosslinking procedure for the formation of the film network. Calcein was used as model active matter for delivery experiments. Multi-Lamellar vesicles (MLV) and Small Uni-Lamellar Vesicles (SUV) were prepared and compared. The liposomes are well distributed throughout the whole area of the film, and the vesicle distribution is equivalent (for both types of liposomes evaluated) on the film surface as well as deeper (100 microns) in the film matrix. An obvious decrease of the burst effect was observed in presence of liposomes as well as a uniform increase of calcein release that continues even at large time scales. Liposomes act as an extra barrier for calcein release. Systems containing MLVs release higher amounts of calcein compared to systems containing SUVs, although these liposomes are more stable in the matrix and diffuse with difficulty. This difference comes from the higher quantity of calcein present within the MLV in relation with their size. Modeling of release kinetics curves was performed and the release of hydrophilic drugs may be described by a multi-scale mechanism characterized by four distinct phases, each of them being characterized by a different kinetics model (Higuchi equation, Korsmeyer-Peppas model etc.). Knowledge of such models will be a very interesting tool for designing new formulations for tissue engineering, regenerative medicine and drug delivery systems.

Keywords: controlled and delayed release, hydrogels, liposomes, polysaccharides

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207 Investigating the Thermal Comfort Properties of Mohair Fabrics

Authors: Adine Gericke, Jiri Militky, Mohanapriya Venkataraman

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Mohair, obtained from the Angora goat, is a luxury fiber and recognized as one of the best quality natural fibers. Expansion of the use of mohair into technical and functional textile products necessitates the need for a better understanding of how the use of mohair in fabrics will impact on its thermo-physiological comfort related properties. Despite its popularity, very little information is available on the quantification of the thermal and moisture management properties of mohair fabrics. This study investigated the effect of fibrous matter composition and fabric structural parameters on conductive and convective heat transfers to attain more information on the thermal comfort properties of mohair fabrics. Dry heat transfer through textiles may involve conduction through the fibrous phase, radiation through fabric interstices and convection of air within the structure. Factors that play a major role in heat transfer by conduction are fabric areal density (g/m2) and derived quantities such as cover factor and porosity. Convective heat transfer through fabrics is found in environmental conditions where there is wind-flow or the object is moving (e.g. running or walking). The thermal comfort properties of mohair fibers were objectively evaluated firstly in comparison with other textile fibers and secondly in a variety of fabric structures. Two sample sets were developed for this purpose, with fibre content, yarn structure and fabric design as main variables. SEM and microscopic images were obtained to closely examine the physical structures of the fibers and fabrics. Thermal comfort properties such as thermal resistance and thermal conductivity, as well as fabric thickness, were measured on the well-known Alambeta test instrument. Clothing insulation (clo) was calculated from the above. The thermal properties of fabrics under heat convection was evaluated using a laboratory model device developed at the Technical University of Liberec (referred to as the TP2-instrument). The effects of the different variables on fabric thermal comfort properties were analyzed statistically using TIBCO Statistica Software. The results showed that fabric structural properties, specifically sample thickness, played a significant role in determining the thermal comfort properties of the fabrics tested. It was found that regarding thermal resistance related to conductive heat flow, the effect of fiber type was not always statistically significant, probably as a result of the amount of trapped air within the fabric structure. The very low thermal conductivity of air, compared to that of the fibers, had a significant influence on the total conductivity and thermal resistance of the samples. This was confirmed by the high correlation of these factors with sample thickness. Regarding convective heat flow, the most important factor influencing the ability of the fabric to allow dry heat to move through the structure, was again fabric thickness. However, it would be wrong to totally disregard the effect of fiber composition on the thermal resistance of textile fabrics. In this study, the samples containing mohair or mohair/wool were consistently thicker than the others even though weaving parameters were kept constant. This can be ascribed to the physical properties of the mohair fibers that renders it exceptionally well towards trapping air among fibers (in a yarn) as well as among yarns (inside a fabric structure). The thicker structures trap more air to provide higher thermal insulation, but also prevent the free flow of air that allow thermal convection.

Keywords: mohair fabrics, convective heat transfer, thermal comfort properties, thermal resistance

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206 Overcoming the Challenges of Subjective Truths in the Post-Truth Age Through a Critical-Ethical English Pedagogy

Authors: Farah Vierra

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Following the 2016 US presidential election and the advancement of the Brexit referendum, the concept of “post-truth,” defined by the Oxford Dictionary as “relating to or denoting circumstances in which objective facts are less influential in shaping public opinion than appeals to emotion and personal belief,” came into prominent use in public, political and educational circles. What this essentially entails is that in this age, individuals are increasingly confronted with subjective perpetuations of truth in their discourse spheres that are informed by beliefs and opinions as opposed to any form of coherence to the reality of those to who this truth claims concern. In principle, a subjective delineation of truth is progressive and liberating – especially considering its potential to provide marginalised groups in the diverse communities of our globalised world with the voice to articulate truths that are representative of themselves and their experiences. However, any form of human flourishing that seems to be promised here collapses as the tenets of subjective truths initially in place to liberate have been distorted through post-truth to allow individuals to purport selective and individualistic truth claims that further oppress and silence certain groups within society without due accountability. The evidence of this is prevalent through the conception of terms such as "alternative facts" and "fake news" that we observe individuals declare when their problematic truth claims are being questioned. Considering the pervasiveness of post-truth and the ethical issues that accompany it, educators and scholars alike have increasingly noted the need to adapt educational practices and pedagogies to account for the diminishing objectivity of truth in the twenty-first century, especially because students, as digital natives, find themselves in the firing line of post-truth; engulfed in digital societies that proliferate post-truth through the surge of truth claims allowed in various media sites. In an attempt to equip students with the vital skills to navigate the post-truth age and oppose its proliferation of social injustices, English educators find themselves having to contend with a complex question: how can the teaching of English equip students with the ability to critically and ethically scrutinise truth claims whilst also mediating the subjectivity of truth in a manner that does not undermine the voices of diverse communities. In order to address this question, this paper will first examine the challenges that confront students as a result of post-truth. Following this, the paper will elucidate the role English education can play in helping students overcome the complex demands of the post-truth age. Scholars have consistently touted the affordances of literary texts in providing students with imagined spaces to explore societal issues through a critical discernment of language and an ethical engagement with its narrative developments. Therefore, this paper will explain and demonstrate how literary texts, when used alongside a critical-ethical post-truth pedagogy that equips students with interpretive strategies informed by literary traditions such as literary and ethical criticism, can be effective in helping students develop the pertinent skills to comprehensively examine truth claims and overcome the challenges of the post-truth age.

Keywords: post-truth, pedagogy, ethics, english, education

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205 Menstruating Bodies and Social Control – Insights From Dignity Without Danger: Collaboratively Analysing Menstrual Stigma and Taboos in Nepal

Authors: Sara Parker, Kay Standing

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This paper will share insights into how menstruators bodies in Nepal are viewed and controlled in Nepal due to the deeply held stigmas and taboos that exist that frame menstrual blood as impure and polluting. It draws on a British Academy Global Challenges Research (BA/GCRF) funded project, ‘Dignity Without Danger,’ that ran from December 2019 to 2022. In Nepal, beliefs and myths around menstrual related practices prevail and vary in accordance to time, generation, caste and class. Physical seclusion and/or restrictions include the consumption of certain foods, the ability to touch certain people and objects, and restricted access to water sources. These restrictions not only put women at risk of poor health outcomes, but they also promote discrimination and challenge fundamental human rights. Despite the pandemic, a wealth of field research and creative outputs have been generated to help break the silence that surrounds menstruation and also highlights the complexity of addressing the harms associated with the exclusion from sacred and profane spaces that menstruators face. Working with locally recruited female research assistants, NGOS and brining together academics from the UK and Nepal, we explore the intersecting factors that impact on menstrual experiences and how they vary throughout Nepal. WE concur with Tamang that there is no such thing as a ‘Nepali Woman’, and there is no one narrative that captures the experiences of menstruators in Nepal. These deeply held beliefs and practices mean that menstruators are denied their right to a dignified menstruation. By being excluded from public and private spaces, such as temples and religious sites, as well as from kitchens and your own bedroom in your own home, these beliefs impact on individuals in complex and interesting ways. Existing research in Nepal by academics and activists demonstrates current programmes and initiatives do not fully address the misconceptions that underpin the exclusionary practices impacting on sexual and reproductive health, a sense of well being and highlight more work is needed in this area. Research has been conducted in all 7 provinces and through exploring and connecting disparate stories, artefacts and narratives, we will deepen understanding of the complexity of menstrual practices enabling local stakeholders to challenge exclusionary practices. By using creative methods to engage with stakeholders and share our research findings as well as highlighting the wealth of activism in Nepal. We highlight the importance of working with local communities, leaders and cutting across disciplines and agencies to promote menstrual justice and dignity. Our research findings and creative outputs that we share on social media channels such as Dignity Without Danger Facebook, Instagram and you tube stress the value of employing a collaborative action research approach to generate material which helps local people take control of their own narrative and change social relations that lead to harmful practices.

Keywords: menstruation, Nepal, stigma, social norms

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204 Evaluating a Peer-To-Peer Health Education Program in Public Housing Communities during the COVID-19 Pandemic

Authors: Jane Oliver, Angeline Ferdinand, Jessica Kaufman, Peta Edler, Nicole Allard, Margie Danchin, Katherine B. Gibney

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Background: The cohealth Health Concierge program operated in Melbourne, Australia, from July 2020 to 30 June 2022. The program was designed to provide place-based peer-to-peer COVID-19 education and support to culturally and linguistically diverse residents of high-rise public housing estates. During this time, the COVID-19 public health response changed frequently. We conducted a mixed-methods evaluation to determine the program’s impact on residents’ trust, engagement and communication with health services and public health activities. Methods: The RE-AIM model was used to assess program reach, effectiveness, adoption, implementation and maintenance and the evaluation was informed by a Project Reference Group including end-users. Data were collected between March and May 2022 in four estates where the program operated. We surveyed 301 residents, conducted qualitative interviews with 32 stakeholders and analyzed data from 20,901 forms reporting interactions between Health Concierges and residents collected from August 2021 to May 2022. These forms outlined the support provided by Health Concierges during each interaction. Results: Overall, the program was effective in guiding residents to testing and vaccination services and facilitating COVID-19 safe practices. Nearly two-thirds (191; 63.5%) of the 301 surveyed participants reported speaking with a Health Concierge in the previous six months, and some described having meaningful conversations with them. Despite this, many of the interactions residents described having with Health Concierges were superficial. When considering surveyed participants’ responses to the adapted Public Health Disaster Trust Scale, the mean score across all estates was 2.3 (or slightly more than ‘somewhat confident’) in public health authorities’ ability to respond to a localized infectious disease outbreak. While the program was valued during the rapidly changing public health response, many felt it had failed to evolve in the ‘living with COVID’ phase. Some residents expressed frustration with Health Concierges’ having perceived inactive, passive roles - although other residents felt Health Concierges were helpful and appreciated them. A perception that the true impact of Health Concierges’ work was underrecognized was widely voiced by health staff. All 20,901 Interaction Forms identified COVID-19-related supports provided to residents; almost all included provision of facemasks and/or hand sanitiser and 78% identified additional supports that were also provided, most frequently provision of other health information. Conclusions: The program disseminated up-to-date information to a diverse population within a rapidly changing public health setting. Health Concierges were able promote COVID-19-safe behaviours, including vaccine uptake, and link residents with support services. We recommend the program be revised and continued. New programs that draw on the Health Concierge model may be valuable in supporting future pandemic responses and should be considered in preparedness planning.

Keywords: community health, COVID-19 pandemic, infectious diseases, public health, community health workers

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203 Sustainable Marine Tourism: Opinion and Segmentation of Italian Generation Z

Authors: M. Bredice, M. B. Forleo, L. Quici

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Coastal tourism is currently facing huge challenges on how to balance environmental problems and tourist activities. Recent literature shows a growing interest in the issue of sustainable tourism from a so-called civilized tourists’ perspective by investigating opinions, perceptions, and behaviors. This study investigates the opinions of youth on what makes them responsible tourists and the ability of coastal marine areas to support tourism in future scenarios. A sample of 778 Italians attending the last year of high school was interviewed. Descriptive statistics, tests, and cluster analyses are applied to highlight the distribution of opinions among youth, detect significant differences based on demographic characteristics, and make segmentation of the different profiles based on students’ opinions and behaviors. Preliminary results show that students are largely convinced (62%) that by 2050 the quality of coastal environments could limit seaside tourism, while 10% of them believe that the problem can be solved simply by changing the tourist destination. Besides the cost of the holiday, the most relevant aspect respondents consider when choosing a marine destination is the presence of tourist attractions followed by the quality of the marine-coastal environment, the specificity of the local gastronomy and cultural traditions, and finally, the activities offered to guests such as sports and events. The reduction of waste and lower air emissions are considered the most important environmental areas in which marine-coastal tourism activities can contribute to preserving the quality of seas and coasts. Areas in which, as a tourist, they believe possible to give a personal contribution were (responses “very much” and “somewhat”); do not throw litter in the sea and on the beach (84%), do not buy single-use plastic products (66%), do not use soap or shampoo when showering in beaches (53%), do not have bonfires (47%), do not damage dunes (46%), and do not remove natural materials (e.g., sand, shells) from the beach (46%). About 6% of the sample stated that they were not interested in contributing to the aforementioned activities, while another 7% replied that they could not contribute at all. Finally, 80% of the sample has never participated in voluntary environmental initiatives or citizen science projects; moreover, about 64% of the students have never participated in events organized by environmental associations in marine or coastal areas. Regarding the test analysis -based on Kruskal-Wallis and Mann and Whitney tests - gender, region, and studying area of students reveals significance in terms of variables expressing knowledge and interest in sustainability topics and sustainable tourism behaviors. The classification of the education field is significant for a great number of variables, among which those related to several sustainable behaviors that respondents declare to be able to contribute as tourists. The ongoing cluster analysis will reveal different profiles in the sample and relevant variables. Based on preliminary results, implications are envisaged in the fields of education, policy, and business strategies for sustainable scenarios. Under these perspectives, the study has the potential to contribute to the conference debate about marine and coastal sustainable development and management.

Keywords: cluster analysis, education, knowledge, young people

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202 Seek First to Regulate, Then to Understand: The Case for Preemptive Regulation of Robots

Authors: Catherine McWhorter

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Robotics is a fast-evolving field lacking comprehensive and harm-mitigating regulation; it also lacks critical data on how human-robot interaction (HRI) may affect human psychology. As most anthropomorphic robots are intended as substitutes for humans, this paper asserts that the commercial robotics industry should be preemptively regulated at the federal level such that robots capable of embodying a victim role in criminal scenarios (“vicbots”) are prohibited until clinical studies determine their effects on the user and society. The results of these studies should then inform more permanent legislation that strives to mitigate risks of harm without infringing upon fundamental rights or stifling innovation. This paper explores these concepts through the lens of the sex robot industry. The sexbot industry offers some of the most realistic, interactive, and customizable robots for sale today. From approximately 2010 until 2017, some sex robot producers, such as True Companion, actively promoted ‘vicbot’ culture with personalities like “Frigid Farrah” and “Young Yoko” but received significant public backlash for fetishizing rape and pedophilia. Today, “Frigid Farrah” and “Young Yoko” appear to have vanished. Sexbot producers have replaced preprogrammed vicbot personalities in favor of one generic, customizable personality. According to the manufacturer ainidoll.com, when asked, there is only one thing the user won’t be able to program the sexbot to do – “…give you drama”. The ability to customize vicbot personas is possible with today’s generic personality sexbots and may undermine the intent of some current legislative efforts. Current debate on the effects of vicbots indicates a lack of consensus. Some scholars suggest vicbots may reduce the rate of actual sex crimes, and some suggest that vicbots will, in fact, create sex criminals, while others cite their potential for rehabilitation. Vicbots may have value in some instances when prescribed by medical professionals, but the overall uncertainty and lack of data further underscore the need for preemptive regulation and clinical research. Existing literature on exposure to media violence and its effects on prosocial behavior, human aggression, and addiction may serve as launch points for specific studies into the hyperrealism of vicbots. Of course, the customization, anthropomorphism and artificial intelligence of sexbots, and therefore more mainstream robots, will continue to evolve. The existing sexbot industry offers an opportunity to preemptively regulate and to research answers to these and many more questions before this type of technology becomes even more advanced and mainstream. Robots pose complicated moral, ethical, and legal challenges, most of which are beyond the scope of this paper. By examining the possibility for custom vicbots via the sexbots industry, reviewing existing literature on regulation, media violence, and vicbot user effects, this paper strives to underscore the need for preemptive federal regulation prohibiting vicbot capabilities in robots while advocating for further research into the potential for the user and societal harm by the same.

Keywords: human-robot interaction effects, regulation, research, robots

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201 Multiple Plant-Based Cell Suspension as a Bio-Ink for 3D Bioprinting Applications in Food Technology

Authors: Yusuf Hesham Mohamed

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Introduction: Three-dimensional printing technology includes multiple procedures that fabricate three-dimensional objects through consecutively layering two-dimensional cross-sections on top of each other. 3D bioprinting is a promising field of 3D printing, which fabricates tissues and organs by accurately controlling the proper arrangement of diverse biological components. 3D bioprinting uses software and prints biological materials and their supporting components layer-by-layer on a substrate or in a tissue culture plate to produce complex live tissues and organs. 3D food printing is an emerging field of 3D bioprinting in which the 3D printed products are food products that are cheap, require less effort to produce, and have more desirable traits. The Aim of the Study is the development of an affordable 3D bioprinter by altering a locally made CNC instrument with an open-source platform to suit the 3D bio-printer purposes. Later, we went through applying the prototype in several applications regarding food technology and drug testing, including the organ-On-Chip. Materials and Methods: An off-the-shelf 3D printer was modified by designing and fabricating the syringe unit, which was designed on the basis of the Milli-fluidics system. Sodium alginate and gelatin hydrogels were prepared, followed by leaf cell suspension preparation from narrow sections of Fragaria’s viable leaves. The desired 3D structure was modeled, and 3D printing preparations took place. Cell-free and cell-laden hydrogels were printed at room temperature under sterile conditions. Post printing curing process was performed. The printed structure was further studied. Results: Positive results have been achieved using the altered 3D bioprinter where a 3D hydrogel construct of two layers made of the combination of sodium alginate to gelatin (15%: 0.5%) has been printed. DLP 3D printer was used to design the syringe component with a transparent PLA-Pro resin for the creation of a microfluidics system having two channels altered to the double extruder. The hydrogel extruder’s design was based on peristaltic pumps, which utilized a stepper motor. The design and fabrication were made using DIY-3D printed parts. Hard plastic PLA was the material utilized for printing. SEM was used to carry out the porous 3D construct imaging. Multiple physical and chemical tests were performed in order to ensure that the cell line was suitable for hosting. Fragaria plant was developed by suspending Fragaria’s cells from its leaves using the 3D bioprinter. Conclusion: 3D bioprinting is considered to be an emerging scientific field that can facilitate and improve many scientific tests and studies. Thus, having a 3D bioprinter in labs is considered to be an essential requirement. 3D bioprinters are very expensive; however, the fabrication of a 3D printer into a 3D bioprinter can lower the cost of the bioprinter. The 3D bioprinter implemented made use of peristaltic pumps instead of syringe-based pumps in order to extend the ability to print multiple types of materials and cells.

Keywords: scaffold, eco on chip, 3D bioprinter, DLP printer

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200 COVID-19’s Impact on the Use of Media, Educational Performance, and Learning in Children and Adolescents with ADHD Who Engaged in Virtual Learning

Authors: Christina Largent, Tazley Hobbs

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Objective: A literature review was performed to examine the existing research on COVID-19 lockdown as it relates to ADHD child/adolescent individuals, media use, and impact on educational performance/learning. It was surmised that with the COVID-19 shut-down and transition to remote learning, a less structured learning environment, increased screen time, in addition to potential difficulty accessing school resources would impair ADHD individuals’ performance and learning. A resulting increase in the number of youths diagnosed and treated for ADHD would be expected. As of yet, there has been little to no published data on the incidence of ADHD as it relates to COVID-19 outside of reports from several nonprofit agencies such as CHADD (Children and Adults with Attention-Deficit/Hyperactivity Disorder ), who reported an increased number of calls to their helpline, The New York based Child Mind Institute, who reported an increased number of appointments to discuss medications, and research released from Athenahealth showing an increase in the number of patients receiving new diagnosis of ADHD and new prescriptions for ADHD medications. Methods: A literature search for articles published between 2020 and 2021 from Pubmed, Google Scholar, PsychInfo, was performed. Search phrases and keywords included “covid, adhd, child, impact, remote learning, media, screen”. Results: Studies primarily utilized parental reports, with very few from the perspective of the ADHD individuals themselves. Most findings thus far show that with the COVID-19 quarantine and transition to online learning, ADHD individuals’ experienced decreased ability to keep focused or adhere to the daily routine, as well as increased inattention-related problems, such as careless mistakes or lack of completion in homework, which in turn translated into overall more difficulty with remote learning. To add further injury, one study showed (just on evaluation of two different sites within the US) that school based services for these individuals decreased with the shift to online-learning. Increased screen time, television, social media, and gaming were noted amongst ADHD individuals. One study further differentiated the degree of digital media, identifying individuals with “problematic “ or “non-problematic” use. ADHD children with problematic digital media use suffered from more severe core symptoms of ADHD, negative emotions, executive function deficits, damage to family environment, pressure from life events, and a lower motivation to learn. Conclusions and Future Considerations: Studies found not only was online learning difficult for ADHD individuals but it, in addition to greater use of digital media, was associated with worsening ADHD symptoms impairing schoolwork, in addition to secondary findings of worsening mood and behavior. Currently, data on the number of new ADHD cases, in addition to data on the prescription and usage of stimulants during COVID-19, has not been well documented or studied; this would be well-warranted out of concern for over diagnosing or over-prescribing our youth. It would also be well-worth studying how reversible or long-lasting these negative impacts may be.

Keywords: COVID-19, remote learning, media use, ADHD, child, adolescent

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199 Fully Instrumented Small-Scale Fire Resistance Benches for Aeronautical Composites Assessment

Authors: Fabienne Samyn, Pauline Tranchard, Sophie Duquesne, Emilie Goncalves, Bruno Estebe, Serge Boubigot

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Stringent fire safety regulations are enforced in the aeronautical industry due to the consequences that potential fire event on an aircraft might imply. This is so much true that the fire issue is considered right from the design of the aircraft structure. Due to the incorporation of an increasing amount of polymer matrix composites in replacement of more conventional materials like metals, the nature of the fire risks is changing. The choice of materials used is consequently of prime importance as well as the evaluation of its resistance to fire. The fire testing is mostly done using the so-called certification tests according to standards such as the ISO2685:1998(E). The latter describes a protocol to evaluate the fire resistance of structures located in fire zone (ability to withstand fire for 5min). The test consists in exposing an at least 300x300mm² sample to an 1100°C propane flame with a calibrated heat flux of 116kW/m². This type of test is time-consuming, expensive and gives access to limited information in terms of fire behavior of the materials (pass or fail test). Consequently, it can barely be used for material development purposes. In this context, the laboratory UMET in collaboration with industrial partners has developed a horizontal and a vertical small-scale instrumented fire benches for the characterization of the fire behavior of composites. The benches using smaller samples (no more than 150x150mm²) enables to cut downs costs and hence to increase sampling throughput. However, the main added value of our benches is the instrumentation used to collect useful information to understand the behavior of the materials. Indeed, measurements of the sample backside temperature are performed using IR camera in both configurations. In addition, for the vertical set up, a complete characterization of the degradation process, can be achieved via mass loss measurements and quantification of the gasses released during the tests. These benches have been used to characterize and study the fire behavior of aeronautical carbon/epoxy composites. The horizontal set up has been used in particular to study the performances and durability of protective intumescent coating on 2mm thick 2D laminates. The efficiency of this approach has been validated, and the optimized coating thickness has been determined as well as the performances after aging. Reductions of the performances after aging were attributed to the migration of some of the coating additives. The vertical set up has enabled to investigate the degradation process of composites under fire. An isotropic and a unidirectional 4mm thick laminates have been characterized using the bench and post-fire analyses. The mass loss measurements and the gas phase analyses of both composites do not present significant differences unlike the temperature profiles in the thickness of the samples. The differences have been attributed to differences of thermal conductivity as well as delamination that is much more pronounced for the isotropic composite (observed on the IR-images). This has been confirmed by X-ray microtomography. The developed benches have proven to be valuable tools to develop fire safe composites.

Keywords: aeronautical carbon/epoxy composite, durability, intumescent coating, small-scale ‘ISO 2685 like’ fire resistance test, X-ray microtomography

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198 Mycophenolate Mofetil Increases Mucin Expression in Primary Cultures of Oral Mucosal Epithelial Cells for Application in Limbal Stem Cell Deficiency

Authors: Sandeep Kumar Agrawal, Aditi Bhattacharya, Janvie Manhas, Krushna Bhatt, Yatin Kholakiya, Nupur Khera, Ajoy Roychoudhury, Sudip Sen

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Autologous cultured explants of human oral mucosal epithelial cells (OMEC) are a potential therapeutic modality for limbal stem cell deficiency (LSCD). Injury or inflammation of the ocular surface in the form of burns, chemicals, Stevens Johnson syndrome, ocular cicatricial pemphigoid etc. can lead to destruction and deficiency of limbal stem cells. LSCD manifests in the form of severe ocular surface diseases (OSD) characterized by persistent and recurrent epithelial defects, conjuntivalisation and neovascularisation of the corneal surface, scarring and ultimately opacity and blindness. Most of the cases of OSD are associated with severe dry eye pertaining to diminished mucin and aqueous secretion. Mycophenolate mofetil (MMF) has been shown to upregulate the mucin expression in conjunctival goblet cells in vitro. The aim of this study was to evaluate the effects of MMF on mucin expression in primary cultures of oral mucosal epithelial cells. With institutional ethics committee approval and written informed consent, thirty oral mucosal epithelial tissue samples were obtained from patients undergoing oral surgery for non-malignant conditions. OMEC were grown on human amniotic membrane (HAM, obtained from expecting mothers undergoing elective caesarean section) scaffold for 2 weeks in growth media containing DMEM & Ham’s F12 (1:1) with 10% FBS and growth factors. In vitro dosage of MMF was standardised by MTT assay. Analysis of stem cell markers was done using RT-PCR while mucin mRNA expression was quantified using RT-PCR and q-PCR before and after treating cultured OMEC with graded concentrations of MMF for 24 hours. Protein expression was validated using immunocytochemistry. Morphological studies revealed a confluent sheet of proliferating, stratified oral mucosal epithelial cells growing over the surface of HAM scaffold. The presence of progenitor stem cell markers (p63, p75, β1-Integrin and ABCG2) and cell surface associated mucins (MUC1, MUC15 and MUC16) were elucidated by RT-PCR. The mucin mRNA expression was found to be upregulated in MMF treated primary cultures of OMEC, compared to untreated controls as quantified by q-PCR with β-actin as internal reference gene. Increased MUC1 protein expression was validated by immunocytochemistry on representative samples. Our findings conclude that OMEC have the ability to form a multi-layered confluent sheet on the surface of HAM similar to a cornea, which is important for the reconstruction of the damaged ocular surface. Cultured OMEC has stem cell properties as demonstrated by stem cell markers. MMF can be a novel enhancer of mucin production in OMEC. It has the potential to improve dry eye in patients undergoing OMEC transplantation for bilateral OSD. Further clinical trials are required to establish the role of MMF in patients undergoing OMEC transplantation.

Keywords: limbal stem cell deficiency, mycophenolate mofetil, mucin, ocular surface disease

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197 Documentary Filmmaking as Activism: Case Studies in Advocacy and Social Justice

Authors: Babatunde Kolawole

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This paper embarks on an exploration of the compelling interplay between documentary filmmaking and activism, delving into their symbiotic relationship and profound impact on advocacy and social justice causes. Through an in-depth analysis of diverse case studies, it seeks to illuminate the instances where documentary films have emerged as potent tools for effecting social change and advancing the principles of justice. This research underscores the vital role played by documentary filmmakers in harnessing the medium's unique capacity to engage, educate, and mobilize audiences while advocating for societal transformation. The primary focus of this study is on a selection of compelling case studies spanning various topics and causes, each exemplifying the marriage between documentary filmmaking and activism. These case studies encompass a broad spectrum of subjects, from environmental conservation and climate change to civil rights movements and human rights struggles. By examining these real-world instances, this paper endeavors to provide a comprehensive understanding of the strategies, challenges, and ethical considerations that underpin the practice of documentary filmmaking as a form of activism. Throughout the paper, it becomes evident that the potency of documentary filmmaking lies in its ability to blend artistry with social impact. The selected case studies vividly demonstrate how documentary filmmakers, armed with cameras and a passion for change, have emerged as critical agents of societal transformation. Whether it be exposing environmental atrocities, shedding light on systemic inequalities, or giving voice to marginalized communities, these documentaries have played a pivotal role in pushing the boundaries of advocacy and social justice. One of the key themes explored in this paper is the evolving nature of documentary filmmaking as a tool for activism. It delves into the shift from traditional observational documentaries to more participatory and immersive approaches, highlighting the dynamic ways in which filmmakers engage with their subjects and audiences. This evolution is exemplified in case studies where filmmakers have collaborated with the communities they document, fostering a sense of agency and empowerment among those whose stories are being told. Furthermore, this research underscores the ethical considerations inherent in the intersection of documentary filmmaking and activism. It scrutinizes questions surrounding representation, objectivity, and the responsibility of filmmakers in portraying complex social issues. By dissecting ethical dilemmas faced by documentary filmmakers in these case studies, this paper encourages a critical examination of the ethical boundaries and obligations in the realm of advocacy-driven filmmaking. In conclusion, this paper aims to shed light on the remarkable potential of documentary filmmaking as a catalyst for activism and social justice. Through the lens of compelling case studies, it illustrates the transformative power of the medium in effecting change, amplifying underrepresented voices, and mobilizing global audiences. It is hoped that this research will not only inform the discourse on documentary activism but also inspire filmmakers, scholars, and advocates to continue leveraging the cinematic art form as a formidable force for a more just and equitable world.

Keywords: film, filmmaker, documentary, human right

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196 Multi-Modality Brain Stimulation: A Treatment Protocol for Tinnitus

Authors: Prajakta Patil, Yash Huzurbazar, Abhijeet Shinde

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Aim: To develop a treatment protocol for the management of tinnitus through multi-modality brain stimulation. Methodology: Present study included 33 adults with unilateral (31 subjects) and bilateral (2 subjects) chronic tinnitus with and/or without hearing loss independent of their etiology. The Treatment protocol included 5 consecutive sessions with follow-up of 6 months. Each session was divided into 3 parts: • Pre-treatment: a) Informed consent b) Pitch and loudness matching. • Treatment: Bimanual paper pen task with tinnitus masking for 30 minutes. • Post-treatment: a) Pitch and loudness matching b) Directive counseling and obtaining feedback. Paper-pen task is to be performed bimanually that included carrying out two different writing activities in different context. The level of difficulty of the activities was increased in successive sessions. Narrowband noise of a frequency same as that of tinnitus was presented at 10 dBSL of tinnitus for 30 minutes simultaneously in the ear with tinnitus. Result: The perception of tinnitus was no longer present in 4 subjects while in remaining subjects it reduced to an intensity that its perception no longer troubled them without causing residual facilitation. In all subjects, the intensity of tinnitus decreased by an extent of 45 dB at an average. However, in few subjects, the intensity of tinnitus also decreased by more than 45 dB. The approach resulted in statistically significant reductions in Tinnitus Functional Index and Tinnitus Handicap Inventory scores. The results correlate with pre and post treatment score of Tinnitus Handicap Inventory that dropped from 90% to 0%. Discussion: Brain mapping(qEEG) Studies report that there is multiple parallel overlapping of neural subnetworks in the non-auditory areas of the brain which exhibits abnormal, constant and spontaneous neural activity involved in the perception of tinnitus with each subnetwork and area reflecting a specific aspect of tinnitus percept. The paper pen task and directive counseling are designed and delivered respectively in a way that is assumed to induce normal, rhythmically constant and premeditated neural activity and mask the abnormal, constant and spontaneous neural activity in the above-mentioned subnetworks and the specific non-auditory area. Counseling was focused on breaking the vicious cycle causing and maintaining the presence of tinnitus. Diverting auditory attention alone is insufficient to reduce the perception of tinnitus. Conscious awareness of tinnitus can be suppressed when individuals engage in cognitively demanding tasks of non-auditory nature as the paper pen task used in the present study. To carry out this task selective, divided, sustained, simultaneous and split attention act cumulatively. Bimanual paper pen task represents a top-down activity which underlies brain’s ability to selectively attend to the bimanual written activity as a relevant stimulus and to ignore tinnitus that is the irrelevant stimuli in the present study. Conclusion: The study suggests that this novel treatment approach is cost effective, time saving and efficient to vanish the tinnitus or to reduce the intensity of tinnitus to a negligible level and thereby eliminating the negative reactions towards tinnitus.

Keywords: multi-modality brain stimulation, neural subnetworks, non-auditory areas, paper-pen task, top-down activity

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195 Investigation of Resilient Circles in Local Community and Industry: Waju-Traditional Culture in Japan and Modern Technology Application

Authors: R. Ueda

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Today global society is seeking resilient partnership in local organizations and individuals, which realizes multi-stakeholders relationship. Although it is proposed by modern global framework of sustainable development, it is conceivable that such affiliation can be found out in the traditional local community in Japan, and that traditional spirit is tacitly sustaining in modern context of disaster mitigation in society and economy. Then this research is aiming to clarify and analyze implication for the global world by actual case studies. Regional and urban resilience is the ability of multi-stakeholders to cooperate flexibly and to adapt in response to changes in the circumstances caused by disasters, but there are various conflicts affecting coordination of disaster relief measures. These conflicts arise not only from a lack of communication and an insufficient network, but also from the difficulty to jointly draw common context from fragmented information. This is because of the weakness of our modern engineering which focuses on maintenance and restoration of individual systems. Here local ‘circles’ holistically includes local community and interacts periodically. Focusing on examples of resilient organizations and wisdom created in communities, what can be seen throughout history is a virtuous cycle where the information and the knowledge are structured, the context to be adapted becomes clear, and an adaptation at a higher level is made possible, by which the collaboration between organizations is deepened and expanded. And the wisdom of a solid and autonomous disaster prevention formed by the historical community called’ Waju’ – an area surrounded by circle embankment to protect the settlement from flood – lives on in government efforts of the coastal industrial island of today. Industrial company there collaborates to create a circle including common evacuation space, road access improvement and infrastructure recovery. These days, people here adopts new interface technology. Large-scale AR- Augmented Reality for more than hundred people is expressing detailed hazard by tsunami and liquefaction. Common experiences of the major disaster space and circle of mutual discussion are enforcing resilience. Collaboration spirit lies in the center of circle. A consistent key point is a virtuous cycle where the information and the knowledge are structured, the context to be adapted becomes clear, and an adaptation at a higher level is made possible, by which the collaboration between organizations is deepened and expanded. This writer believes that both self-governing human organizations and the societal implementation of technical systems are necessary. Infrastructure should be autonomously instituted by associations of companies and other entities in industrial areas for working closely with local governments. To develop advanced disaster prevention and multi-stakeholder collaboration, partnerships among industry, government, academia and citizens are important.

Keywords: industrial recovery, multi-sakeholders, traditional culture, user experience, Waju

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194 Investigating the Association between Escherichia Coli Infection and Breast Cancer Incidence: A Retrospective Analysis and Literature Review

Authors: Nadia Obaed, Lexi Frankel, Amalia Ardeljan, Denis Nigel, Anniki Witter, Omar Rashid

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Breast cancer is the most common cancer among women, with a lifetime risk of one in eight of all women in the United States. Although breast cancer is prevalent throughout the world, the uneven distribution in incidence and mortality rates is shaped by the variation in population structure, environment, genetics and known lifestyle risk factors. Furthermore, the bacterial profile in healthy and cancerous breast tissue differs with a higher relative abundance of bacteria capable of causing DNA damage in breast cancer patients. Previous bacterial infections may change the composition of the microbiome and partially account for the environmental factors promoting breast cancer. One study found that higher amounts of Staphylococcus, Bacillus, and Enterobacteriaceae, of which Escherichia coli (E. coli) is a part, were present in breast tumor tissue. Based on E. coli’s ability to damage DNA, it is hypothesized that there is an increased risk of breast cancer associated with previous E. coli infection. Therefore, the purpose of this study was to evaluate the correlation between E. coli infection and the incidence of breast cancer. Holy Cross Health, Fort Lauderdale, provided access to the Health Insurance Portability and Accountability (HIPAA) compliant national database for the purpose of academic research. International Classification of Disease 9th and 10th Codes (ICD-9, ICD-10) was then used to conduct a retrospective analysis using data from January 2010 to December 2019. All breast cancer diagnoses and all patients infected versus not infected with E. coli that underwent typical E. coli treatment were investigated. The obtained data were matched for age, Charlson Comorbidity Score (CCI score), and antibiotic treatment. Standard statistical methods were applied to determine statistical significance and an odds ratio was used to estimate the relative risk. A total of 81286 patients were identified and analyzed from the initial query and then reduced to 31894 antibiotic-specific treated patients in both the infected and control group, respectively. The incidence of breast cancer was 2.51% and present in 2043 patients in the E. coli group compared to 5.996% and present in 4874 patients in the control group. The incidence of breast cancer was 3.84% and present in 1223 patients in the treated E. coli group compared to 6.38% and present in 2034 patients in the treated control group. The decreased incidence of breast cancer in the E. coli and treated E. coli groups was statistically significant with a p-value of 2.2x10-16 and 2.264x10-16, respectively. The odds ratio in the E. coli and treated E. coli groups was 0.784 and 0.787 with a 95% confidence interval, respectively (0.756-0.813; 0.743-0.833). The current study shows a statistically significant decrease in breast cancer incidence in association with previous Escherichia coli infection. Researching the relationship between single bacterial species is important as only up to 10% of breast cancer risk is attributable to genetics, while the contribution of environmental factors including previous infections potentially accounts for a majority of the preventable risk. Further evaluation is recommended to assess the potential and mechanism of E. coli in decreasing the risk of breast cancer.

Keywords: breast cancer, escherichia coli, incidence, infection, microbiome, risk

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193 On-Farm Biopurification Systems: Fungal Bioaugmentation of Biomixtures For Carbofuran Removal

Authors: Carlos E. Rodríguez-Rodríguez, Karla Ruiz-Hidalgo, Kattia Madrigal-Zúñiga, Juan Salvador Chin-Pampillo, Mario Masís-Mora, Elizabeth Carazo-Rojas

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One of the main causes of contamination linked to agricultural activities is the spillage and disposal of pesticides, especially during the loading, mixing or cleaning of agricultural spraying equipment. One improvement in the handling of pesticides is the use of biopurification systems (BPS), simple and cheap degradation devices where the pesticides are biologically degraded at accelerated rates. The biologically active core of BPS is the biomixture, which is constituted by soil pre-exposed to the target pesticide, a lignocellulosic substrate to promote the activity of ligninolitic fungi and a humic component (peat or compost), mixed at a volumetric proportion of 50:25:25. Considering the known ability of lignocellulosic fungi to degrade a wide range of organic pollutants, and the high amount of lignocellulosic waste used in biomixture preparation, the bioaugmentation of biomixtures with these fungi represents an interesting approach for improving biomixtures. The present work aimed at evaluating the effect of the bioaugmentation of rice husk based biomixtures with the fungus Trametes versicolor in the removal of the insectice/nematicide carbofuran (CFN) and to optimize the composition of the biomixture to obtain the best performance in terms of CFN removal and mineralization, reduction in formation of transformation products and decrease in residual toxicity of the matrix. The evaluation of several lignocellulosic residues (rice husk, wood chips, coconut fiber, sugarcane bagasse or newspaper print) revealed the best colonization by T. versicolor in rice husk. Pre-colonized rice husk was then used in the bioaugmentation of biomixtures also containing soil pre-exposed to CFN and either peat (GTS biomixture) or compost (GCS biomixture). After spiking with 10 mg/kg CBF, the efficiency of the biomixture was evaluated through a multi-component approach that included: monitoring of CBF removal and production of CBF transformation products, mineralization of radioisotopically labeled carbofuran (14C-CBF) and changes in the toxicity of the matrix after the treatment (Daphnia magna acute immobilization test). Estimated half-lives of CBF in the biomixtures were 3.4 d and 8.1 d in GTS and GCS, respectively. The transformation products 3-hydroxycarbofuran and 3-ketocarbofuran were detected at the moment of CFN application, however their concentration continuously disappeared. Mineralization of 14C-CFN was also faster in GTS than GCS. The toxicological evaluation showed a complete toxicity removal in the biomixtures after 48 d of treatment. The composition of the GCS biomixture was optimized using a central composite design and response surface methodology. The design variables were the volumetric content of fungally pre-colonized rice husk and the volumetric ratio compost/soil. According to the response models, maximization of CFN removal and mineralization rate, and minimization in the accumulation of transformation products were obtained with an optimized biomixture of composition 30:43:27 (pre-colonized rice husk:compost:soil), which differs from the 50:25:25 composition commonly employed in BPS. Results suggest that fungal bioaugmentation may enhance the performance of biomixtures in CFN removal. Optimization reveals the importance of assessing new biomixture formulations in order to maximize their performance.

Keywords: bioaugmentation, biopurification systems, degradation, fungi, pesticides, toxicity

Procedia PDF Downloads 287
192 Rethinking the Languages for Specific Purposes Syllabus in the 21st Century: Topic-Centered or Skills-Centered

Authors: A. Knezović

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21st century has transformed the labor market landscape in a way of posing new and different demands on university graduates as well as university lecturers, which means that the knowledge and academic skills students acquire in the course of their studies should be applicable and transferable from the higher education context to their future professional careers. Given the context of the Languages for Specific Purposes (LSP) classroom, the teachers’ objective is not only to teach the language itself, but also to prepare students to use that language as a medium to develop generic skills and competences. These include media and information literacy, critical and creative thinking, problem-solving and analytical skills, effective written and oral communication, as well as collaborative work and social skills, all of which are necessary to make university graduates more competitive in everyday professional environments. On the other hand, due to limitations of time and large numbers of students in classes, the frequently topic-centered syllabus of LSP courses places considerable focus on acquiring the subject matter and specialist vocabulary instead of sufficient development of skills and competences required by students’ prospective employers. This paper intends to explore some of those issues as viewed both by LSP lecturers and by business professionals in their respective surveys. The surveys were conducted among more than 50 LSP lecturers at higher education institutions in Croatia, more than 40 HR professionals and more than 60 university graduates with degrees in economics and/or business working in management positions in mainly large and medium-sized companies in Croatia. Various elements of LSP course content have been taken into consideration in this research, including reading and listening comprehension of specialist texts, acquisition of specialist vocabulary and grammatical structures, as well as presentation and negotiation skills. The ability to hold meetings, conduct business correspondence, write reports, academic texts, case studies and take part in debates were also taken into consideration, as well as informal business communication, business etiquette and core courses delivered in a foreign language. The results of the surveys conducted among LSP lecturers will be analyzed with reference to what extent those elements are included in their courses and how consistently and thoroughly they are evaluated according to their course requirements. Their opinions will be compared to the results of the surveys conducted among professionals from a range of industries in Croatia so as to examine how useful and important they perceive the same elements of the LSP course content in their working environments. Such comparative analysis will thus show to what extent the syllabi of LSP courses meet the demands of the employment market when it comes to the students’ language skills and competences, as well as transferable skills. Finally, the findings will also be compared to the observations based on practical teaching experience and the relevant sources that have been used in this research. In conclusion, the ideas and observations in this paper are merely open-ended questions that do not have conclusive answers, but might prompt LSP lecturers to re-evaluate the content and objectives of their course syllabi.

Keywords: languages for specific purposes (LSP), language skills, topic-centred syllabus, transferable skills

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191 Molecular Migration in Polyvinyl Acetate Matrix: Impact of Compatibility, Number of Migrants and Stress on Surface and Internal Microstructure

Authors: O. Squillace, R. L. Thompson

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Migration of small molecules to, and across the surface of polymer matrices is a little-studied problem with important industrial applications. Tackifiers in adhesives, flavors in foods and binding agents in paints all present situations where the function of a product depends on the ability of small molecules to migrate through a polymer matrix to achieve the desired properties such as softness, dispersion of fillers, and to deliver an effect that is felt (or tasted) on a surface. It’s been shown that the chemical and molecular structure, surface free energies, phase behavior, close environment and compatibility of the system, influence the migrants’ motion. When differences in behavior, such as occurrence of segregation to the surface or not, are observed it is then of crucial importance to identify and get a better understanding of the driving forces involved in the process of molecular migration. In this aim, experience is meant to be allied with theory in order to deliver a validated theoretical and computational toolkit to describe and predict these phenomena. The systems that have been chosen for this study aim to address the effect of polarity mismatch between the migrants and the polymer matrix and that of a second migrant over the first one. As a non-polar resin polymer, polyvinyl acetate is used as the material to which more or less polar migrants (sorbitol, carvone, octanoic acid (OA), triacetin) are to be added. Through contact angle measurement a surface excess is seen for sorbitol (polar) mixed with PVAc as the surface energy is lowered compare to the one of pure PVAc. This effect is increased upon the addition of carvon or triacetin (non-polars). Surface micro-structures are also evidenced by atomic force microscopy (AFM). Ion beam analysis (Nuclear Reaction Analysis), supplemented by neutron reflectometry can accurately characterize the self-organization of surfactants, oligomers, aromatic molecules in polymer films in order to relate the macroscopic behavior to the length scales that are amenable to simulation. The nuclear reaction analysis (NRA) data for deuterated OA 20% shows the evidence of a surface excess which is enhanced after annealing. The addition of 10% triacetin, as a second migrant, results in the formation of an underlying layer enriched in triacetin below the surface excess of OA. The results show that molecules in polarity mismatch with the matrix tend to segregate to the surface, and this is favored by the addition of a second migrant of the same polarity than the matrix. As studies have been restricted to materials that are model supported films under static conditions in a first step, it is also wished to address the more challenging conditions of materials under controlled stress or strain. To achieve this, a simple rig and PDMS cell have been designed to stretch the material to a defined strain and to probe these mechanical effects by ion beam analysis and atomic force microscopy. This will make a significant step towards exploring the influence of extensional strain on surface segregation, flavor release in cross-linked rubbers.

Keywords: polymers, surface segregation, thin films, molecular migration

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190 Targeting Peptide Based Therapeutics: Integrated Computational and Experimental Studies of Autophagic Regulation in Host-Parasite Interaction

Authors: Vrushali Guhe, Shailza Singh

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Cutaneous leishmaniasis is neglected tropical disease present worldwide caused by the protozoan parasite Leishmania major, the therapeutic armamentarium for leishmaniasis are showing several limitations as drugs are showing toxic effects with increasing resistance by a parasite. Thus identification of novel therapeutic targets is of paramount importance. Previous studies have shown that autophagy, a cellular process, can either facilitate infection or aid in the elimination of the parasite, depending on the specific parasite species and host background in leishmaniasis. In the present study, our objective was to target the essential autophagy protein ATG8, which plays a crucial role in the survival, infection dynamics, and differentiation of the Leishmania parasite. ATG8 in Leishmania major and its homologue, LC3, in Homo sapiens, act as autophagic markers. Present study manifested the crucial role of ATG8 protein as a potential target for combating Leishmania major infection. Through bioinformatics analysis, we identified non-conserved motifs within the ATG8 protein of Leishmania major, which are not present in LC3 of Homo sapiens. Against these two non-conserved motifs, we generated a peptide library of 60 peptides on the basis of physicochemical properties. These peptides underwent a filtering process based on various parameters, including feasibility of synthesis and purification, compatibility with Selective Reaction Monitoring (SRM)/Multiple reaction monitoring (MRM), hydrophobicity, hydropathy index, average molecular weight (Mw average), monoisotopic molecular weight (Mw monoisotopic), theoretical isoelectric point (pI), and half-life. Further filtering criterion shortlisted three peptides by using molecular docking and molecular dynamics simulations. The direct interaction between ATG8 and the shortlisted peptides was confirmed through Surface Plasmon Resonance (SPR) experiments. Notably, these peptides exhibited the remarkable ability to penetrate the parasite membrane and exert profound effects on Leishmania major. The treatment with these peptides significantly impacted parasite survival, leading to alterations in the cell cycle and morphology. Furthermore, the peptides were found to modulate autophagosome formation, particularly under starved conditions, suggesting their involvement in disrupting the regulation of autophagy within Leishmania major. In vitro, studies demonstrated that the selected peptides effectively reduced the parasite load within infected host cells. Encouragingly, these findings were corroborated by in vivo experiments, which showed a reduction in parasite burden upon peptide administration. Additionally, the peptides were observed to affect the levels of LC3II within host cells. In conclusion, our findings highlight the efficacy of these novel peptides in targeting Leishmania major’s ATG8 and disrupting parasite survival. These results provide valuable insights into the development of innovative therapeutic strategies against leishmaniasis via targeting autophagy protein ATG8 of Leishmania major.

Keywords: ATG8, leishmaniasis, surface plasmon resonance, MD simulation, molecular docking, peptide designing, therapeutics

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189 Stakeholder Mapping and Requirements Identification for Improving Traceability in the Halal Food Supply Chain

Authors: Laila A. H. F. Dashti, Tom Jackson, Andrew West, Lisa Jackson

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Traceability systems are important in the agri-food and halal food sectors for monitoring ingredient movements, tracking sources, and ensuring food integrity. However, designing a traceability system for the halal food supply chain is challenging due to diverse stakeholder requirements and complex needs. Existing literature on stakeholder mapping and identifying requirements for halal food supply chains is limited. To address this gap, a pilot study was conducted to identify the objectives, requirements, and recommendations of stakeholders in the Kuwaiti halal food industry. The study collected data through semi-structured interviews with an international halal food manufacturer based in Kuwait. The aim was to gain a deep understanding of stakeholders' objectives, requirements, processes, and concerns related to the design of a traceability system in the country's halal food sector. Traceability systems are being developed and tested in the agri-food and halal food sectors due to their ability to monitor ingredient movements, track sources, and detect potential issues related to food integrity. Designing a traceability system for the halal food supply chain poses significant challenges due to diverse stakeholder requirements and the complexity of their needs (including varying food ingredients, different sources, destinations, supplier processes, certifications, etc.). Achieving a halal food traceability solution tailored to stakeholders' requirements within the supply chain necessitates prior knowledge of these needs. Although attempts have been made to address design-related issues in traceability systems, literature on stakeholder mapping and identification of requirements specific to halal food supply chains is scarce. Thus, this pilot study aims to identify the objectives, requirements, and recommendations of stakeholders in the halal food industry. The paper presents insights gained from the pilot study, which utilized semi-structured interviews to collect data from a Kuwait-based international halal food manufacturer. The objective was to gain an in-depth understanding of stakeholders' objectives, requirements, processes, and concerns pertaining to the design of a traceability system in Kuwait's halal food sector. The stakeholder mapping results revealed that government entities, food manufacturers, retailers, and suppliers are key stakeholders in Kuwait's halal food supply chain. Lessons learned from this pilot study regarding requirement capture for traceability systems include the need to streamline communication, focus on communication at each level of the supply chain, leverage innovative technologies to enhance process structuring and operations and reduce halal certification costs. The findings also emphasized the limitations of existing traceability solutions, such as limited cooperation and collaboration among stakeholders, high costs of implementing traceability systems without government support, lack of clarity regarding product routes, and disrupted communication channels between stakeholders. These findings contribute to a broader research program aimed at developing a stakeholder requirements framework that utilizes "business process modelling" to establish a unified model for traceable stakeholder requirements.

Keywords: supply chain, traceability system, halal food, stakeholders’ requirements

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188 Effect of Radioprotectors on DNA Repair Enzyme and Survival of Gamma-Irradiated Cell Division Cycle Mutants of Saccharomyces pombe

Authors: Purva Nemavarkar, Badri Narain Pandey, Jitendra Kumar

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Introduction: The objective was to understand the effect of various radioprotectors on DNA damage repair enzyme and survival in gamma-irradiated wild and cdc mutants of S. pombe (fission yeast) cultured under permissive and restrictive conditions. DNA repair process, as influenced by radioprotectors, was measured by activity of DNA polymerase in the cells. The use of single cell gel electrophoresis assay (SCGE) or Comet Assay to follow gamma-irradiation induced DNA damage and effect of radioprotectors was employed. In addition, studying the effect of caffeine at different concentrations on S-phase of cell cycle was also delineated. Materials and Methods: S. pombe cells grown at permissive temperature (250C) and/or restrictive temperature (360C) were followed by gamma-radiation. Percentage survival and activity of DNA Polymerase (yPol II) were determined after post-irradiation incubation (5 h) with radioprotectors such as Caffeine, Curcumin, Disulphiram, and Ellagic acid (the dose depending on individual D 37 values). The gamma-irradiated yeast cells (with and without the radioprotectors) were spheroplasted by enzyme glusulase and subjected to electrophoresis. Radio-resistant cells were obtained by arresting cells in S-phase using transient treatment of hydroxyurea (HU) and studying the effect of caffeine at different concentrations on S-phase of cell cycle. Results: The mutants of S. pombe showed insignificant difference in survival when grown under permissive conditions. However, growth of these cells under restrictive temperature leads to arrest in specific phases of cell cycle in different cdc mutants (cdc10: G1 arrest, cdc22: early S arrest, cdc17: late S arrest, cdc25: G2 arrest). All the cdc mutants showed decrease in survival after gamma radiation when grown at permissive and restrictive temperatures. Inclusion of the radioprotectors at respective concentrations during post irradiation incubation showed increase in survival of cells. Activity of DNA polymerase enzyme (yPol II) was increased significantly in cdc mutant cells exposed to gamma-radiation. Following SCGE, a linear relationship was observed between doses of irradiation and the tail moments of comets. The radioprotection of the fission yeast by radioprotectors can be seen by the reduced tail moments of the yeast comets. Caffeine also exhibited its radio-protective ability in radio-resistant S-phase cells obtained after HU treatment. Conclusions: The radioprotectors offered notable radioprotection in cdc mutants when added during irradiation. The present study showed activation of DNA damage repair enzyme (yPol II) and an increase in survival after treatment of radioprotectors in gamma irradiated wild type and cdc mutants of S. pombe cells. Results presented here showed feasibility of applying SCGE in fission yeast to follow DNA damage and radioprotection at high doses, which are not feasible with other eukaryotes. Inclusion of caffeine at 1mM concentration to S phase cells offered protection and did not decrease the cell viability. It can be proved that at minimal concentration, caffeine offered marked radioprotection.

Keywords: radiation protection, cell cycle, fission yeast, comet assay, s-phase, DNA repair, radioprotectors, caffeine, curcumin, SCGE

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187 Tele-Rehabilitation for Multiple Sclerosis: A Case Study

Authors: Sharon Harel, Rachel Kizony, Yoram Feldman, Gabi Zeilig, Mordechai Shani

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Multiple Sclerosis (MS) is a neurological disease that may cause restriction in participation in daily activities of young adults. Main symptoms include fatigue, weakness and cognitive decline. The appearance of symptoms, their severity and deterioration rate, change between patients. The challenge of health services is to provide long-term rehabilitation services to people with MS. The objective of this presentation is to describe a course of tele-rehabilitation service of a woman with MS. Methods; R is a 48 years-old woman, diagnosed with MS when she was 22. She started to suffer from weakness of her non-dominant left upper extremity about ten years after the diagnosis. She was referred to the tele-rehabilitation service by her rehabilitation team, 16 years after diagnosis. Her goals were to improve ability to use her affected upper extremity in daily activities. On admission her score in the Mini-Mental State Exam was 30/30. Her Fugl-Meyer Assessment (FMA) score of the left upper extremity was 48/60, indicating mild weakness and she had a limitation of her shoulder abduction (90 degrees). In addition, she reported little use of her arm in daily activities as shown in her responses to the Motor Activity Log (MAL) that were equal to 1.25/5 in amount and 1.37 in quality of use. R. received two 30 minutes on-line sessions per week in the tele-rehabilitation service, with the CogniMotion system. These were complemented by self-practice with the system. The CogniMotion system provides a hybrid (synchronous-asynchronous), the home-based tele-rehabilitation program to improve the motor, cognitive and functional status of people with neurological deficits. The system consists of a computer, large monitor, and the Microsoft’s Kinect 3D sensor. This equipment is located in the client’s home and connected to a clinician’s computer setup in a remote clinic via WiFi. The client sits in front of the monitor and uses his body movements to interact with games and tasks presented on the monitor. The system provides feedback in the form of ‘knowledge of results’ (e.g., the success of a game) and ‘knowledge of performance’ (e.g., alerts for compensatory movements) to enhance motor learning. The games and tasks were adapted for R. motor abilities and level of difficulty was gradually increased according to her abilities. The results of her second assessment (after 35 on-line sessions) showed improvement in her FMA score to 52 and shoulder abduction to 140 degrees. Moreover, her responses to the MAL indicated an increased amount (2.4) and quality (2.2) of use of her left upper extremity in daily activities. She reported high level of enjoyment from the treatments (5/5), specifically the combination of cognitive challenges while moving her body. In addition, she found the system easy to use as reflected by her responses to the System Usability Scale (85/100). To-date, R. continues to receive treatments in the tele-rehabilitation service. To conclude, this case report shows the potential of using tele-rehabilitation for people with MS to provide strategies to enhance the use of the upper extremity in daily activities as well as for maintaining motor function.

Keywords: motor function, multiple-sclerosis, tele-rehabilitation, daily activities

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186 Designing Next Generation Platforms for Recombinant Protein Production by Genome Engineering of Escherichia coli

Authors: Priyanka Jain, Ashish K. Sharma, Esha Shukla, K. J. Mukherjee

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We propose a paradigm shift in our approach to design improved platforms for recombinant protein production, by addressing system level issues rather than the individual steps associated with recombinant protein synthesis like transcription, translation, etc. We demonstrate that by controlling and modulating the cellular stress response (CSR), which is responsible for feedback control of protein synthesis, we can generate hyper-producing strains. We did transcriptomic profiling of post-induction cultures, expressing different types of protein, to analyze the nature of this cellular stress response. We found significant down-regulation of substrate utilization, translation, and energy metabolism genes due to generation CSR inside the host cell. However, transcription profiling has also shown that many genes are up-regulated post induction and their role in modulating the CSR is unclear. We hypothesized that these up-regulated genes trigger signaling pathways, generating the CSR and concomitantly reduce the recombinant protein yield. To test this hypothesis, we knocked out the up-regulated genes, which did not have any downstream regulatees, and analyzed their impact on cellular health and recombinant protein expression. Two model proteins i.e., GFP and L-Asparaginase were chosen for this analysis. We observed a significant improvement in expression levels, with some knock-outs showing more than 7-fold higher expression compared to control. The 10 best single knock-outs were chosen to make 45 combinations of all possible double knock-outs. A further increase in expression was observed in some of these double knock- outs with GFP levels being highest in a double knock-out ΔyhbC + ΔelaA. However, for L-Asparaginase which is a secretory protein, the best results were obtained using a combination of ΔelaA+ΔcysW knock-outs. We then tested all the knock outs for their ability to enhance the expression of a 'difficult-to-express' protein. The Rubella virus E1 protein was chosen and tagged with sfGFP at the C-terminal using a linker peptide for easy online monitoring of expression of this fusion protein. Interestingly, the highest increase in Rubella-sGFP levels was obtained in the same double knock-out ΔelaA + ΔcysW (5.6 fold increase in expression yield compared to the control) which gave the highest expression for L-Asparaginase. However, for sfGFP alone, the ΔyhbC+ΔmarR knock-out gave the highest level of expression. These results indicate that there is a fair degree of commonality in the nature of the CSR generated by the induction of different proteins. Transcriptomic profiling of the double knock out showed that many genes associated with the translational machinery and energy biosynthesis did not get down-regulated post induction, unlike the control where these genes were significantly down-regulated. This confirmed our hypothesis of these genes playing an important role in the generation of the CSR and allowed us to design a strategy for making better expression hosts by simply knocking out key genes. This strategy is radically superior to the previous approach of individually up-regulating critical genes since it blocks the mounting of the CSR thus preventing the down-regulation of a very large number of genes responsible for sustaining the flux through the recombinant protein production pathway.

Keywords: cellular stress response, GFP, knock-outs, up-regulated genes

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185 Bacteriophages for Sustainable Wastewater Treatment: Application in Black Water Decontamination with an Emphasis to DRDO Biotoilet

Authors: Sonika Sharma, Mohan G. Vairale, Sibnarayan Datta, Soumya Chatterjee, Dharmendra Dubey, Rajesh Prasad, Raghvendra Budhauliya, Bidisha Das, Vijay Veer

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Bacteriophages are viruses that parasitize specific bacteria and multiply in metabolising host bacteria. Bacteriophages hunt for a single or a subset of bacterial species, making them potential antibacterial agents. Utilizing the ability of phages to control bacterial populations has several applications from medical to the fields of agriculture, aquaculture and the food industry. However, harnessing phage based techniques in wastewater treatments to improve quality of effluent and sludge release into the environment is a potential area for R&D application. Phage mediated bactericidal effect in any wastewater treatment process has many controlling factors that lead to treatment performance. In laboratory conditions, titer of bacteriophages (coliphages) isolated from effluent water of a specially designed anaerobic digester of human night soil (DRDO Biotoilet) was successfully increased with a modified protocol of the classical double layer agar technique. Enrichment of the same was carried out and efficacy of the phage enriched medium was evaluated at different conditions (specific media, temperature, storage conditions). Growth optimization study was carried out on different media like soybean casein digest medium (Tryptone soya medium), Luria-Bertani medium, phage deca broth medium and MNA medium (Modified nutrient medium). Further, temperature-phage yield relationship was also observed at three different temperatures 27˚C, 37˚C and 44˚C at laboratory condition. Results showed the higher activity of coliphage 27˚C and at 37˚C. Further, addition of divalent ions (10mM MgCl2, 5mM CaCl2) and 5% glycerol resulted in a significant increase in phage titer. Besides this, effect of antibiotics addition like ampicillin and kanamycin at different concentration on plaque formation was analysed and reported that ampicillin at a concentration of 1mg/ml ampicillin stimulates phage infection and results in more number of plaques. Experiments to test viability of phage showed that it can remain active for 6 months at 4˚C in fresh tryptone soya broth supplemented with fresh culture of coliforms (early log phase). The application of bacteriophages (especially coliphages) for treatment of effluent of human faecal matter contaminated effluent water is unique. This environment-friendly treatment system not only reduces the pathogenic coliforms, but also decreases the competition between nuisance bacteria and functionally important microbial populations. Therefore, the phage based cocktail to treat fecal pathogenic bacteria present in black water has many implication in wastewater treatment processes including ‘DRDO Biotoilet’, which is an ecofriendly appropriate and affordable human faecal matter treatment technology for different climates and situations.

Keywords: wastewater, microbes, virus, biotoilet, phage viability

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184 The Effect of Metabolites of Fusarium solani on the Activity of the PR-Proteins (Chitinase, β-1,3-Glucanase and Peroxidases) of Potato Tubers

Authors: A. K. Tursunova, O. V. Chebonenko, A. Zh. Amirkulova, A. O. Abaildayev, O. A. Sapko, Y. M. Dyo, A. Sh. Utarbaeva

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Fusarium solani and its variants cause root and stem rot of plants. Dry rot is the most common disease of potato tubers during storage. The causative agents of fusariosis in contact with plants behave as antagonists, growth stimulants or parasites. The diversity of host-parasite relationships is explained by the parasite’s ability to produce a wide spectrum of biologically active compounds including toxins, enzymes, oligosaccharides, antibiotic substances, enniatins and gibberellins. Many of these metabolites contribute to the creation of compatible relations; others behave as elicitors, inducing various protective responses in plants. An important part of the strategy for developing plant resistance against pathogens is the activation of protein synthesis to produce protective ‘pathogenesis-related’ proteins. The family of PR-proteins known to confer the most protective response is chitinases (EC 3.2.1.14, Cht) and β-1,3-glucanases (EC 3.2.1.39, Glu). PR-proteins also include a large multigene family of peroxidases (EC 1.11.1.7, Pod), and increased activity of Pod and expression of the Pod genes leads to the development of resistance to a broad class of pathogens. Despite intensive research on the role of PR-proteins, the question of their participation in the mechanisms of formation of the F.solani–S.tuberosum pathosуstem is not sufficiently studied. Our aim was to investigate the effect of different classes of F. solani metabolites on the activity of chitinase, β-1,3-glucanases and peroxidases in tubers of Solanum tuberosum. Metabolite culture filtrate (CF) and cytoplasmic components were fractionated by extraction of the mycelium with organic solvents, salting out techniques, dialysis, column chromatography and ultrafiltration. Protein, lipid, carbohydrate and polyphenolic fractions of fungal metabolites were derived. Using enzymatic hydrolysis we obtained oligo glycans from fungal cell walls with different molecular weights. The activity of the metabolites was tested using potato tuber discs (d = 16mm, h = 5mm). The activity of PR-proteins of tubers was analyzed in a time course of 2–24 hours. The involvement of the analysed metabolites in the modulation of both early non-specific and late related to pathogenesis reactions was demonstrated. The most effective inducer was isolated from the CF (fraction of total phenolic compounds including naphtazarins). Induction of PR-activity by this fraction was: chitinase - 340-360%, glucanase - 435-450%, soluble forms of peroxidase - 400-560%, related forms of peroxidase - 215-237%. High-inducing activity was observed by the chloroform and acetonitrile extracts of the mycelium (induction of chitinase and glucanase activity was 176-240%, of soluble and bound forms of peroxidase - 190-400%). The fraction of oligo glycans mycelium cell walls of 1.2 kDa induced chitinase and β-1,3-glucanase to 239-320%; soluble forms and related peroxidase to 198-426%. Oligo glycans cell walls of 5-10 kDa had a weak suppressor effect - chitinase (21-25%) and glucanase (25-28%) activity; had no effect on soluble forms of peroxidase, but induced to 250-270% activity related forms. The CF polysaccharides of 8.5 kDa and 3.1 kDa inhibited synchronously the glucanase and chitinase specific response in step (after 24 hours at 42-50%) and the step response induced nonspecific peroxidase activity: soluble forms 4.8 -5.2 times, associated forms 1.4-1.6 times.

Keywords: fusarium solani, PR-proteins, peroxidase, solanum tuberosum

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183 Experimental-Numerical Inverse Approaches in the Characterization and Damage Detection of Soft Viscoelastic Layers from Vibration Test Data

Authors: Alaa Fezai, Anuj Sharma, Wolfgang Mueller-Hirsch, André Zimmermann

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Viscoelastic materials have been widely used in the automotive industry over the last few decades with different functionalities. Besides their main application as a simple and efficient surface damping treatment, they may ensure optimal operating conditions for on-board electronics as thermal interface or sealing layers. The dynamic behavior of viscoelastic materials is generally dependent on many environmental factors, the most important being temperature and strain rate or frequency. Prior to the reliability analysis of systems including viscoelastic layers, it is, therefore, crucial to accurately predict the dynamic and lifetime behavior of these materials. This includes the identification of the dynamic material parameters under critical temperature and frequency conditions along with a precise damage localization and identification methodology. The goal of this work is twofold. The first part aims at applying an inverse viscoelastic material-characterization approach for a wide frequency range and under different temperature conditions. For this sake, dynamic measurements are carried on a single lap joint specimen using an electrodynamic shaker and an environmental chamber. The specimen consists of aluminum beams assembled to adapter plates through a viscoelastic adhesive layer. The experimental setup is reproduced in finite element (FE) simulations, and frequency response functions (FRF) are calculated. The parameters of both the generalized Maxwell model and the fractional derivatives model are identified through an optimization algorithm minimizing the difference between the simulated and the measured FRFs. The second goal of the current work is to guarantee an on-line detection of the damage, i.e., delamination in the viscoelastic bonding of the described specimen during frequency monitored end-of-life testing. For this purpose, an inverse technique, which determines the damage location and size based on the modal frequency shift and on the change of the mode shapes, is presented. This includes a preliminary FE model-based study correlating the delamination location and size to the change in the modal parameters and a subsequent experimental validation achieved through dynamic measurements of specimen with different, pre-generated crack scenarios and comparing it to the virgin specimen. The main advantage of the inverse characterization approach presented in the first part resides in the ability of adequately identifying the material damping and stiffness behavior of soft viscoelastic materials over a wide frequency range and under critical temperature conditions. Classic forward characterization techniques such as dynamic mechanical analysis are usually linked to limitations under critical temperature and frequency conditions due to the material behavior of soft viscoelastic materials. Furthermore, the inverse damage detection described in the second part guarantees an accurate prediction of not only the damage size but also its location using a simple test setup and outlines; therefore, the significance of inverse numerical-experimental approaches in predicting the dynamic behavior of soft bonding layers applied in automotive electronics.

Keywords: damage detection, dynamic characterization, inverse approaches, vibration testing, viscoelastic layers

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182 Antioxidant Potency of Ethanolic Extracts from Selected Aromatic Plants by in vitro Spectrophotometric Analysis

Authors: Tatjana Kadifkova Panovska, Svetlana Kulevanova, Blagica Jovanova

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Biological systems possess the ability to neutralize the excess of reactive oxygen species (ROS) and to protect cells from destructive alterations. However, many pathological conditions (cardiovascular diseases, autoimmune disorders, cancer) are associated with inflammatory processes that generate an excessive amount of reactive oxygen species (ROS) that shift the balance between endogenous antioxidant systems and free oxygen radicals in favor of the latter, leading to oxidative stress. Therefore, an additional source of natural compounds with antioxidant properties that will reduce the amount of ROS in cells is much needed despite their broad utilization; many plant species remain largely unexplored. Therefore, the purpose of the present study is to investigate the antioxidant activity of twenty-five selected medicinal and aromatic plant species. The antioxidant activity of the ethanol extracts was evaluated with in vitro assays: 2,2’-diphenyl-1-pycryl-hydrazyl (DPPH), ferric reducing antioxidant power (FRAP), non-site-specific- (NSSOH) and site-specific hydroxyl radical-2-deoxy-D-ribose degradation (SSOH) assays. The Folin-Ciocalteu method and AlCl3 method were performed to determine total phenolic content (TPC) and total flavonoid content (TFC). All examined plant extracts manifested antioxidant activity to a different extent. Cinnamomum verum J.Presl bark and Ocimum basilicum L. Herba demonstrated strong radical scavenging activity and reducing power with the DPPH and FRAP assay, respectively. Additionally, significant hydroxyl scavenging potential and metal chelating properties were observed using the NSSOH and SSOH assays. Furthermore, significant variations were determined in the total polyphenolic content (TPC) and total flavonoid content (TFC), with Cinnamomum verum and Ocimum basilicum showing the highest amount of total polyphenols. The considerably strong radical scavenging activity, hydroxyl scavenging potential and reducing power for the species mentioned above suggest of a presence of highly bioactive phytochemical compounds, predominantly polyphenols. Since flavonoids are the most abundant group of polyphenols that possess a large number of available reactive OH groups in their structure, it is considered that they are the main contributors to the radical scavenging properties of the examined plant extracts. This observation is supported by the positive correlation between the radical scavenging activity and the total polyphenolic and flavonoid content obtained in the current research. The observations from the current research nominate Cinnamomum verum bark and Ocimum basilicum herba as potential sources of bioactive compounds that could be utilized as antioxidative additives in the food and pharmaceutical industries. Moreover, the present study will help the researchers as basic data for future research in exploiting the hidden potential of these important plants that have not been explored so far.

Keywords: ethanol extracts, radical scavenging activity, reducing power, total polyphenols.

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