Search results for: allergic contact dermatitis
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 1965

Search results for: allergic contact dermatitis

75 Quality Assessment of Pedestrian Streets in Iran: Case Study of Saf, Tehran

Authors: Fstemeh Rais Esmaili, Ehsan Ranjbar

Abstract:

Pedestrian streets as one type of urban public spaces have an important role in improving the quality of urban life. In Iran, planning and designing of pedestrian streets is in its primary steps. In spite of starting this approach in Iran, and designing several pedestrian streets, there are still not organized studies about quality assessment of pedestrian streets. As a result, the strength and weakness points of the initial experiences have not been utilized. This inattention to quality assessment have caused designing pedestrian streets to be limited to just vehicles traffic control and preliminary actions like paving; so that, special potentials of pedestrian streets for creating social, livable and dynamic public spaces have not been used. This article, as an organized study about quality assessment of pedestrian streets in Iran, tries to reach two main goals: first, introducing a framework for quality assessment of pedestrian streets in Iran, and second, creating a context for improving the quality of pedestrian streets especially for further experiences. The main research methods are description and context analyzing. With respect to comparative analysis of ideas about quality, considering international and local case studies and analyzing existing condition of Saf Pedestrian Street, a particular model for quality assessment has been introduced. In this model, main components and assessment criteria have been presented. On the basis of this model, questionnaire and checklist for assessment have been prepared. The questionnaire and interview have been used to assess qualities which are in direct contact with people and the checklist has been used for analyzing visual qualities by authors through observation. Some results of questionnaire and checklist show that 7 of 11 primary components, diversity, flexibility, cleanness, legibility and imaginably, identity, livability, form and physical setting are rated low and very low in quality degree. Three components, efficiency, comfort and distinctiveness, have medium and low quality degree and one component, access, linkage and permeability has high quality degree. Therefore, based on implemented analyzing process, Saf Pedestrian Street needs to be improved and these quality improvement priorities are determined based on presented criteria. Adaption of final results with existing condition illustrates the shortage of services for satisfying user’s needs, inflexibility and impossibility of using spaces in various times, lack of facilities for different climatic conditions, lack of facilities such as drinking fountain, inappropriate designing of existing urban furniture like garbage cans, and creating pollution and unsuitable view, lack of visual attractions, neglecting disabled persons in designing entrances, shortage of benches and their undesirable designing, lack of vegetation, absence of special characters making it different from other streets, preventing people taking part in the space causing lack of affiliation, lack of appropriate elements for leisure time and lack of exhilaration in the space. On the other hand, these results present high access and permeability, high safety, less sound pollution and more relief, comfortable movement along the way due to suitable pavement and economic efficiency, as the strength points of Saf pedestrian street.

Keywords: pedestrian streets, quality assessment, quality criteria, Saf Pedestrian Street

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74 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

Abstract:

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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73 The Lessons Learned from Managing Malignant Melanoma During COVID-19 in a Plastic Surgery Unit in Ireland

Authors: Amenah Dhannoon, Ciaran Martin Hurley, Laura Wrafter, Podraic J. Regan

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Introduction: The COVID-19 pandemic continues to present unprecedented challenges for healthcare systems. This has resulted in the pragmatic shift in the practice of plastic surgery units worldwide. During this period, many units reported a significant fall in urgent melanoma referrals, leading to patients presenting with advanced disease requiring more extensive surgery and inferior outcomes. Our objective was to evaluate our unit's experience with both non-invasive and invasive melanoma during the COVID-19 pandemic and characterize our experience and contrast it to that experienced by our neighbors in the UK, mainland Europe and North America. Methods: a retrospective chart review was performed on all patients diagnosed with invasive and non-invasive cutaneous melanoma between March to December of 2019 (control) compared to 2020 (COVID-19 pandemic) in a single plastic surgery unit in Ireland. Patient demographics, referral source, surgical procedures, tumour characteristics, radiological findings, oncological therapies and follow-up were recorded. All data were anonymized and stored in Microsoft Excel. Results: A total of 589 patients were included in the study. Of these, 314 (53%) with invasive melanoma, compared to 275 (47%) with the non-invasive disease. Overall, more patients were diagnosed with both invasive and non-invasive melanoma in 2020 than in 2019 (p<0.05). However, significantly longer waiting times in 2020 (64 days) compared to 2019 (28 days) (p<0.05), with the majority of the referral being from GP in 2019 (83%) compared to 61% in 2020. Positive sentinel lymph node were higher in 2019 at 56% (n=28) compared to 24% (n=22) in 2020. There was no statistically significant difference in the tutor characteristics or metastasis status. Discussion: While other countries have noticed a fall in the melanoma diagnosis. Our units experienced a higher number of disease diagnoses. This can be due to multiple reasons. In Ireland, the government reached an early agreement with the private sector to continue elective surgery on an urgent basis in private hospitals. This allowed access to local anesthetic procedures and local skin cancer cases were triaged to non-COVID-19 provider centers. Our unit also adapted a fast, effective and minimal patient contact strategy for triaging skin cancer based on telemedicine. Thirdly, a skin cancer nurse specialist maintained patient follow-ups and triaging a dedicated email service. Finally, our plastic surgery service continued to maintain a virtual complex skin cancer multidisciplinary team meeting during the pandemic, ensuring local clinical governance has adhered to each clinical case. Conclusion: Our study highlights that with the prompt efficient restructuring of services, we could reserve successful management of skin cancer even in the most devastating times. It is important to reflect on the success during the pandemic and emphasize the importance of preparation for a potentially difficult future

Keywords: malignant melanoma, skin cancer, COVID-19, triage

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72 Construction and Cross-Linking of Polyelectrolyte Multilayers Based on Polysaccharides as Antifouling Coatings

Authors: Wenfa Yu, Thuva Gnanasampanthan, John Finlay, Jessica Clarke, Charlotte Anderson, Tony Clare, Axel Rosenhahn

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Marine biofouling is a worldwide problem at vast economic and ecological costs. Historically it was combated with toxic coatings such as tributyltin. As those coatings being banned nowadays, finding environmental friendly antifouling solution has become an urgent topic. In this study antifouling coatings consisted of natural occurring polysaccharides hyaluronic acid (HA), alginic acid (AA), chitosan (Ch) and polyelectrolyte polyethylenimine (PEI) are constructed into polyelectrolyte multilayers (PEMs) in a Layer-by-Layer (LbL) method. LbL PEM construction is a straightforward way to assemble biomacromolecular coatings on surfaces. Advantages about PEM include ease of handling, highly diverse PEM composition, precise control over the thickness and so on. PEMs have been widely employed in medical application and there are numerous studies regarding their protein adsorption, elasticity and cell adhesive properties. With the adjustment of coating composition, termination layer charge, coating morphology and cross-linking method, it is possible to prepare low marine biofouling coatings with PEMs. In this study, using spin coating technology, PEM construction was achieved at smooth multilayers with roughness as low as 2nm rms and highly reproducible thickness around 50nm. To obtain stability in sea water, the multilayers were covalently cross-linked either thermally or chemically. The cross-linking method affected surface energy, which was reflected in water contact angle, thermal cross-linking led to hydrophobic surfaces and chemical cross-linking generated hydrophilic surfaces. The coatings were then evaluated regarding its protein resistance and biological species resistance. While the hydrophobic thermally cross-linked PEM had low resistance towards proteins, the resistance of chemically cross-linked PEM strongly depended on the PEM termination layer and the charge of the protein, opposite charge caused high adsorption and same charge low adsorption, indicating electrostatic interaction plays a crucial role in the protein adsorption processes. Ulva linza was chosen as the biological species for antifouling performance evaluation. Despite of the poor resistance towards protein adsorption, thermally cross-linked PEM showed good resistance against Ulva spores settlement, the chemically cross-linked multilayers showed poor resistance regardless of the termination layer. Marine species adhesion is a complex process, although it involves proteins as bioadhesives, protein resistance its own is not a fully indicator for its antifouling performance. The species will pre select the surface, responding to cues like surface energy, chemistry, or charge and so on. Thus making it difficult for one single factors to determine its antifouling performance. Preparing PEM coating is a comprehensive work involving choosing polyelectrolyte combination, determining termination layer and the method for cross-linking. These decisions will affect PEM properties such as surface energy, charge, which is crucial, since biofouling is a process responding to surface properties in a highly sensitive and dynamic way.

Keywords: hyaluronic acid, polyelectrolyte multilayers, protein resistance, Ulva linza zoospores

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71 University Climate and Psychological Adjustment: African American Women’s Experiences at Predominantly White Institutions in the United States

Authors: Faheemah N. Mustafaa, Tamarie Macon, Tabbye Chavous

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A major concern of university leaders worldwide is how to create environments where students from diverse racial/ethnic, national, and cultural backgrounds can thrive. Over the past decade or so in the United States, African American women have done exceedingly well in terms of college enrollment, academic performance, and completion. However, the relative academic successes of African American women in higher education has in some ways overshadowed social challenges many Black women continue to encounter on college campuses in the United States. Within predominantly White institutions (PWIs) in particular, there is consistent evidence that many Black students experience racially hostile climates. However, research studies on racial climates within PWIs have mostly focused on cross-sectional comparisons of minority and majority group experiences, and few studies have examined campus racial climate in relation to short- and longer-term well-being. One longitudinal study reported that African American women’s psychological well-being was positively related to their comfort in cross-racial interactions (a concept closely related to campus climate). Thus, our primary research question was: Do African American women’s perceptions of campus climate (tension and positive association) during their freshman year predict their reports of psychological distress and well-being (self-acceptance) during their sophomore year? Participants were part of a longitudinal survey examining African American college students’ academic identity development, particularly in Science, Technology, Engineering, and Mathematics (STEM) fields. The final subsample included 134 self-identified African American/Black women enrolled in PWIs. Accounting for background characteristics (mother’s education, family income, interracial contact, and prior levels of outcomes), we employed hierarchical regression to examine relationships between campus racial climate during freshman year and psychological adjustment one year later. Both regression models significantly predicted African American women’s psychological outcomes (for distress, F(7,91)= 4.34, p < .001; and for self-acceptance, F(7,90)= 4.92, p < .001). Although none of the controls were significant predictors, perceptions of racial tension on campus were associated with both distress and self-acceptance. More perceptions of tension were related to African American women’s greater psychological distress the following year (B= 0.22, p= .01). Additionally, racial tension predicted later self-acceptance in the expected direction: Higher first-year reports of racial tension were related to less positive attitudes toward the self during the sophomore year (B= -0.16, p= .04). However, perceptions that it was normative for Black and White students to socialize on campus (or positive association scores) were unrelated to psychological distress or self-acceptance. Findings highlight the relevance of examining multiple facets of campus racial climate in relation to psychological adjustment, with possible emphasis on the import of racial tension on African American women’s psychological adjustment. Results suggest that negative dimensions of campus racial climate may have lingering effects on psychological well-being, over and above more positive aspects of climate. Thus, programs targeted toward improving student relations on campus should consider addressing cross-racial tensions.

Keywords: higher education, psychological adjustment, university climate, university students

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70 Development of PCL/Chitosan Core-Shell Electrospun Structures

Authors: Hilal T. Sasmazel, Seda Surucu

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Skin tissue engineering is a promising field for the treatment of skin defects using scaffolds. This approach involves the use of living cells and biomaterials to restore, maintain, or regenerate tissues and organs in the body by providing; (i) larger surface area for cell attachment, (ii) proper porosity for cell colonization and cell to cell interaction, and (iii) 3-dimensionality at macroscopic scale. Recent studies on this area mainly focus on fabrication of scaffolds that can closely mimic the natural extracellular matrix (ECM) for creation of tissue specific niche-like environment at the subcellular scale. Scaffolds designed as ECM-like architectures incorporating into the host with minimal scarring/pain and facilitate angiogenesis. This study is related to combining of synthetic PCL and natural chitosan polymers to form 3D PCL/Chitosan core-shell structures for skin tissue engineering applications. Amongst the polymers used in tissue engineering, natural polymer chitosan and synthetic polymer poly(ε-caprolactone) (PCL) are widely preferred in the literature. Chitosan has been among researchers for a very long time because of its superior biocompatibility and structural resemblance to the glycosaminoglycan of bone tissue. However, the low mechanical flexibility and limited biodegradability properties reveals the necessity of using this polymer in a composite structure. On the other hand, PCL is a versatile polymer due to its low melting point (60°C), ease of processability, degradability with non-enzymatic processes (hydrolysis) and good mechanical properties. Nevertheless, there are also several disadvantages of PCL such as its hydrophobic structure, limited bio-interaction and susceptibility to bacterial biodegradation. Therefore, it became crucial to use both of these polymers together as a hybrid material in order to overcome the disadvantages of both polymers and combine advantages of those. The scaffolds here were fabricated by using electrospinning technique and the characterizations of the samples were done by contact angle (CA) measurements, scanning electron microscopy (SEM), transmission electron microscopy (TEM) and X-Ray Photoelectron spectroscopy (XPS). Additionally, gas permeability test, mechanical test, thickness measurement and PBS absorption and shrinkage tests were performed for all type of scaffolds (PCL, chitosan and PCL/chitosan core-shell). By using ImageJ launcher software program (USA) from SEM photographs the average inter-fiber diameter values were calculated as 0.717±0.198 µm for PCL, 0.660±0.070 µm for chitosan and 0.412±0.339 µm for PCL/chitosan core-shell structures. Additionally, the average inter-fiber pore size values exhibited decrease of 66.91% and 61.90% for the PCL and chitosan structures respectively, compare to PCL/chitosan core-shell structures. TEM images proved that homogenous and continuous bead free core-shell fibers were obtained. XPS analysis of the PCL/chitosan core-shell structures exhibited the characteristic peaks of PCL and chitosan polymers. Measured average gas permeability value of produced PCL/chitosan core-shell structure was determined 2315±3.4 g.m-2.day-1. In the future, cell-material interactions of those developed PCL/chitosan core-shell structures will be carried out with L929 ATCC CCL-1 mouse fibroblast cell line. Standard MTT assay and microscopic imaging methods will be used for the investigation of the cell attachment, proliferation and growth capacities of the developed materials.

Keywords: chitosan, coaxial electrospinning, core-shell, PCL, tissue scaffold

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69 Parents’ Perceptions of the Consent Arrangements for Dental Public Health Programmes in North London: A Qualitative Exploration

Authors: Charlotte Jeavons, Charitini Stavropoulous, Nicolas Drey

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Background: Over one-third of five-year-olds and almost half of all eight-year-olds in the UK have obvious caries experience that can be detected by visual screening techniques. School-based caries preventions programs to apply fluoride varnish to young children’s teeth operate in many areas in the UK. Their aim is to reduce dental caries in children. The Department of Health guidance (2009) on consent states information must be provided to parents to enable informed autonomous decision-making prior to any treatment involving their young children. Fluoride varnish schemes delivered in primary schools use letters for this purpose. Parents are expected to return these indicating their consent or refusal. A large proportion of parents do not respond. In the absence of positive consent, these children are excluded from the program. Non-response is more common in deprived areas creating inequality. The reason for this is unknown. The consent process used is underpinned by the ethical theory of deontology that is prevalent in clinical dentistry and widely accepted in bio-ethics. Objective: To investigate parents’ views, understanding and experience of the fluoride varnish program taking place in their child’s school, including their views about the practical consent arrangements. Method: Schools participating in the fluoride varnish scheme operating in Enfield, North London, were asked to take part. Parents with children in nursery, reception, or year one were invited to participate via semi-structured interviews and focus groups. Thematic analysis was conducted. Findings: 40 parents were recruited from eight schools. The global theme of ‘trust’ was identified as the strongest influence on parental responses. Six themes were identified; protecting children from harm is viewed by parents as their role, parents have the capability to decide but lack confidence, sharing responsibility for their child’s oral health with the State is welcomed by a parent, existing relationships within parents’ social networks strongly influences consent decisions, official dental information is not communicated effectively, sending a letter to parents’ and excluding them from meeting dental practitioners is ineffective. The information delivered via a letter was not strongly identified by parents as influencing their response. Conclusions: Personal contact with the person(s) providing information and requesting consent has a greater impact on parental consent responses than written information provided alone. This demonstrates that traditional bio-ethical ideas about rational decision-making where emotions are transcended and interference is not justified unless preventing harm to an unaware person are outdated. Parental decision-making is relational and the consent process should be adapted to reflect this. The current system that has a deontology view of decision making at its core impoverishes parental autonomy and may, ultimately, increase dental inequalities as a result.

Keywords: consent, decision, ethics, fluoride, parents

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68 Maternal Obesity in Nigeria: An Exploratory Study

Authors: Ojochenemi J. Onubi, Debbi Marais, Lorna Aucott, Friday Okonofua, Amudha Poobalan

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Background: Obesity is a worldwide epidemic with major health and economic consequences. Pregnancy is a trigger point for the development of obesity, and maternal obesity is associated with significant adverse effects in the mother and child. Nigeria is experiencing a double burden of under- and over-nutrition with rising levels of obesity particularly in women. However, there is scarcity of data on maternal obesity in Nigeria and other African countries. Aims and Objectives: This project aimed at identifying crucial components of potential interventions for maternal obesity in Nigeria. The objectives were to assess the prevalence, effects, and distribution of maternal obesity; knowledge, attitude and practice (KAP) of pregnant women and maternal healthcare providers; and identify existing interventions for maternal obesity in Nigeria. Methodology: A systematic review and meta-analysis were initially conducted to appraise the existing literature on maternal obesity in Africa. Following this, a quantitative questionnaire survey of the KAP of pregnant women and a qualitative interview study of the KAP of Health Care Workers (HCW) were conducted in seven secondary and tertiary hospitals across Nigeria. Quantitative data was analysed using SPSS statistical software, while thematic analysis was conducted for qualitative data. Results: Twenty-nine studies included in the systematic review showed significant prevalence, socio-demographic associations, and adverse effects of maternal obesity on labour, maternal, and child outcomes in Africa. The questionnaire survey of 435 mothers revealed a maternal obesity prevalence of 17.9% among mothers who registered for antenatal care in the first trimester. The mothers received nutrition information from different sources and had insufficient knowledge of their own weight category or recommended Gestational Weight Gain (GWG), causes, complications, and safe ways to manage maternal obesity. However, majority of the mothers were of the opinion that excess GWG is avoided in pregnancy and some practiced weight management (diet and exercise) during pregnancy. For the qualitative study, four main themes were identified: ‘Concerns about obesity in pregnancy’, ‘Barriers to care for obese pregnant women’, ‘Practice of care for obese pregnant women’, and ‘Improving care for obese pregnant women’. HCW expressed concerns about rising levels of maternal obesity, lack of guidelines for the management of obese pregnant women and worries about unintended consequences of antenatal interventions. ‘Barriers’ included lack of contact with obese women before pregnancy, late registration for antenatal care, and perceived maternal barriers such as socio-cultural beliefs of mothers and poverty. ‘Practice’ included anticipatory care and screening for possible complications, general nutrition education during antenatal care and interdisciplinary care for mothers with complications. HCW offered suggestions on improving care for obese women including timing, type, and settings of interventions; and the need for involvement of other stake holders in caring for obese pregnant women. Conclusions: Culturally adaptable/sensitive interventions should be developed for the management of obese pregnant women in Africa. Education and training of mothers and health care workers, and provision of guidelines are some of the components of potential interventions in Nigeria.

Keywords: Africa, maternal, obesity, pregnancy

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67 Growth Patterns of Pyrite Crystals Studied by Electron Back Scatter Diffraction (EBSD)

Authors: Kirsten Techmer, Jan-Erik Rybak, Simon Rudolph

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Natural formed pyrites (FeS2) are frequent sulfides in sedimentary and metamorphic rocks. Growth textures of idiomorphic pyrite assemblages reflect the conditions during their formation in the geologic sequence, furtheron the local texture analyses of the growth patterns of pyrite assemblages by EBSD reveal the possibility to resolve the growth conditions during the formation of pyrite at the micron scale. The spatial resolution of local texture measurements in the Scanning Electron Microscope used can be in the nanomete scale. Orientation contrasts resulting from domains of smaller misorientations within larger pyrite crystals can be resolved as well. The electron optical studies have been carried out in a Field-Emission Scanning Electron Microscope (FEI Quanta 200) equipped with a CCD camera to study the orientation contrasts along the surfaces of pyrite. Idiomorphic cubic single crystals of pyrite, polycrystalline assemblages of pyrite, spherically grown spheres of pyrite as well as pyrite-bearing ammonites have been studied by EBSD in the Scanning Electron Microscope. Samples were chosen to show no or minor secondary deformation and an idiomorphic 3D crystal habit, so the local textures of pyrite result mainly from growth and minor from deformation. The samples studied derived from Navajun (Spain), Chalchidiki (Greece), Thüringen (Germany) and Unterkliem (Austria). Chemical analyses by EDAX show pyrite with minor inhomogeneities e.g., single crystals of galena and chalcopyrite along the grain boundaries of larger pyrite crystals. Intergrowth between marcasite and pyrite can be detected in one sample. Pyrite may form intense growth twinning lamellae on {011}. Twinning, e.g., contact twinning is abundant within the crystals studied and the individual twinning lamellaes can be resolved by EBSD. The ammonites studied show a replacement of the shale by newly formed pyrite resulting in an intense intergrowth of calcite and pyrite. EBSD measurements indicate a polycrystalline microfabric of both minerals, still reflecting primary surface structures of the ammonites e.g, the Septen. Discs of pyrite (“pyrite dollar”) as well as pyrite framboids show growth patterns comprising a typical microfabric. EBSD studies reveal an equigranular matrix in the inner part of the discs of pyrite and a fiber growth with larger misorientations in the outer regions between the individual segments. This typical microfabric derived from a formation of pyrite crystals starting at a higher nucleation rate and followed by directional crystal growth. EBSD studies show, that the growth texture of pyrite in the samples studied reveals a correlation between nucleation rate and following growth rate of the pyrites, thus leading to the characteristic crystal habits. Preferential directional growth at lower nucleation rates may lead to the formation of 3D framboids of pyrite. Crystallographic misorientations between the individual fibers are similar. In ammonites studied, primary anisotropies of the substrates like e.g., ammonitic sutures, influence the nucleation, crystal growth and habit of the newly formed pyrites along the surfaces.

Keywords: Electron Back Scatter Diffraction (EBSD), growth pattern, Fe-sulfides (pyrite), texture analyses

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66 Smart and Active Package Integrating Printed Electronics

Authors: Joana Pimenta, Lorena Coelho, José Silva, Vanessa Miranda, Jorge Laranjeira, Rui Soares

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In this paper, the results of R&D on an innovative food package for increased shelf-life are presented. SAP4MA aims at the development of a printed active device that enables smart packaging solutions for food preservation, targeting the extension of the shelf-life of the packed food through the controlled release of active natural antioxidant agents at the onset of the food degradation process. To do so, SAP4MA focuses on the development of active devices such as printed heaters and batteries/supercapacitors in a label format to be integrated on packaging lids during its injection molding process, promoting the passive release of natural antioxidants after the product is packed, during transportation and in the shelves, and actively when the end-user activates the package, just prior to consuming the product at home. When the active device present on the lid is activated, the release of the natural antioxidants embedded in the inner layer of the packaging lid in direct contact with the headspace atmosphere of the food package starts. This approach is based on the use of active functional coatings composed of nano encapsulated active agents (natural antioxidants species) in the prevention of the oxidation of lipid compounds in food by agents such as oxygen. Thus keeping the product quality during the shelf-life, not only when the user opens the packaging, but also during the period from food packaging up until the purchase by the consumer. The active systems that make up the printed smart label, heating circuit, and battery were developed using screen-printing technology. These systems must operate under the working conditions associated with this application. The printed heating circuit was studied using three different substrates and two different conductive inks. Inks were selected, taking into consideration that the printed circuits will be subjected to high pressures and temperatures during the injection molding process. The circuit must reach a homogeneous temperature of 40ºC in the entire area of the lid of the food tub, promoting a gradual and controlled release of the antioxidant agents. In addition, the circuit design involves a high level of study in order to guarantee maximum performance after the injection process and meet the specifications required by the control electronics component. Furthermore, to characterize the different heating circuits, the electrical resistance promoted by the conductive ink and the circuit design, as well as the thermal behavior of printed circuits on different substrates, were evaluated. In the injection molding process, the serpentine-shaped design developed for the heating circuit was able to resolve the issues connected to the injection point; in addition, the materials used in the support and printing had high mechanical resistance against the pressure and temperature inherent to the injection process. Acknowledgment: This research has been carried out within the Project “Smart and Active Packing for Margarine Product” (SAP4MA) running under the EURIPIDES Program being co-financed by COMPETE 2020 – the Operational Programme for Competitiveness and Internationalization and under Portugal 2020 through the European Regional Development Fund (ERDF).

Keywords: smart package, printed heat circuits, printed batteries, flexible and printed electronic

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65 Quantum Chemical Prediction of Standard Formation Enthalpies of Uranyl Nitrates and Its Degradation Products

Authors: Mohamad Saab, Florent Real, Francois Virot, Laurent Cantrel, Valerie Vallet

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All spent nuclear fuel reprocessing plants use the PUREX process (Plutonium Uranium Refining by Extraction), which is a liquid-liquid extraction method. The organic extracting solvent is a mixture of tri-n-butyl phosphate (TBP) and hydrocarbon solvent such as hydrogenated tetra-propylene (TPH). By chemical complexation, uranium and plutonium (from spent fuel dissolved in nitric acid solution), are separated from fission products and minor actinides. During a normal extraction operation, uranium is extracted in the organic phase as the UO₂(NO₃)₂(TBP)₂ complex. The TBP solvent can form an explosive mixture called red oil when it comes in contact with nitric acid. The formation of this unstable organic phase originates from the reaction between TBP and its degradation products on the one hand, and nitric acid, its derivatives and heavy metal nitrate complexes on the other hand. The decomposition of the red oil can lead to violent explosive thermal runaway. These hazards are at the origin of several accidents such as the two in the United States in 1953 and 1975 (Savannah River) and, more recently, the one in Russia in 1993 (Tomsk). This raises the question of the exothermicity of reactions that involve TBP and all other degradation products, and calls for a better knowledge of the underlying chemical phenomena. A simulation tool (Alambic) is currently being developed at IRSN that integrates thermal and kinetic functions related to the deterioration of uranyl nitrates in organic and aqueous phases, but not of the n-butyl phosphate. To include them in the modeling scheme, there is an urgent need to obtain the thermodynamic and kinetic functions governing the deterioration processes in liquid phase. However, little is known about the thermodynamic properties, like standard enthalpies of formation, of the n-butyl phosphate molecules and of the UO₂(NO₃)₂(TBP)₂ UO₂(NO₃)₂(HDBP)(TBP) and UO₂(NO₃)₂(HDBP)₂ complexes. In this work, we propose to estimate the thermodynamic properties with Quantum Methods (QM). Thus, in the first part of our project, we focused on the mono, di, and tri-butyl complexes. Quantum chemical calculations have been performed to study several reactions leading to the formation of mono-(H₂MBP), di-(HDBP), and TBP in gas and liquid phases. In the gas phase, the optimal structures of all species were optimized using the B3LYP density functional. Triple-ζ def2-TZVP basis sets were used for all atoms. All geometries were optimized in the gas-phase, and the corresponding harmonic frequencies were used without scaling to compute the vibrational partition functions at 298.15 K and 0.1 Mpa. Accurate single point energies were calculated using the efficient localized LCCSD(T) method to the complete basis set limit. Whenever species in the liquid phase are considered, solvent effects are included with the COSMO-RS continuum model. The standard enthalpies of formation of TBP, HDBP, and H2MBP are finally predicted with an uncertainty of about 15 kJ mol⁻¹. In the second part of this project, we have investigated the fundamental properties of three organic species that mostly contribute to the thermal runaway: UO₂(NO₃)₂(TBP)₂, UO₂(NO₃)₂(HDBP)(TBP), and UO₂(NO₃)₂(HDBP)₂ using the same quantum chemical methods that were used for TBP and its derivatives in both the gas and the liquid phase. We will discuss the structures and thermodynamic properties of all these species.

Keywords: PUREX process, red oils, quantum chemical methods, hydrolysis

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64 The Prevalence of Soil Transmitted Helminths among Newly Arrived Expatriate Labors in Jeddah, Saudi Arabia

Authors: Mohammad Al-Refai, Majed Wakid

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Introduction: Soil-transmitted diseases (STD) are caused by intestinal worms that are transmitted via various routes into the human body resulting in various clinical manifestations. The intestinal worms causing these infections are known as soil transmitted helminths (STH), including Hook worms, Ascaris lumbricoides (A. lumbricoides), Trichuris trichiura (T. trichiura), and Strongyloides sterocoralis (S. sterocoralis). Objectives: The aim of this study was to investigate the prevalence of STH among newly arrived expatriate labors in Jeddah city, Saudi Arabia, using three different techniques (direct smears, sedimentation concentration, and real-time PCR). Methods: A total of 188 stool specimens were collected and investigated at the parasitology laboratory in the Special Infectious Agents Unit at King Fahd Medical Research Center, King Abdulaziz University in Jeddah, Saudi Arabia. Microscopic examination of wet mount preparations using normal saline and Lugols Iodine was carried out, followed by the formal ether sedimentation method. In addition, real-time PCR was used as a molecular tool to detect several STH and hookworm speciation. Results: Out of 188 stool specimens analyzed, in addition to STH parasite, several other types were detected. 9 samples (4.79%) were positive for Entamoeba coli, 7 samples (3.72%) for T. trichiura, 6 samples (3.19%) for Necator americanus, 4 samples (2.13%) for S. sterocoralis, 4 samples (2.13%) for A. lumbricoides, 4 samples (2.13%) for E. histolytica, 3 samples (1.60%) for Blastocystis hominis, 2 samples (1.06%) for Ancylostoma duodenale, 2 samples (1.06%) for Giardia lamblia, 1 sample (0.53%) for Iodamoeba buetschlii, 1 sample (0.53%) for Hymenolepis nana, 1 sample (0.53%) for Endolimax nana, and 1 sample (0.53%) for Heterophyes heterophyes. Out of the 35 infected cases, 26 revealed single infection, 8 with double infections, and only one triple infection of different STH species and other intestinal parasites. Higher rates of STH infections were detected among housemaids (11 cases) followed by drivers (7 cases) when compared to other occupations. According to educational level, illiterate participants represent the majority of infected workers (12 cases). The majority of workers' positive cases were from the Philippines. In comparison between laboratory techniques, out of the 188 samples screened for STH, real-time PCR was able to detect the DNA in (19/188) samples followed by Ritchie sedimentation technique (18/188), and direct wet smear (7/188). Conclusion: STH infections are a major public health issue to healthcare systems around the world. Communities must be educated on hygiene practices and the severity of such parasites to human health. As far as drivers and housemaids come to close contact with families, including children and elderlies. This may put family members at risk of developing serious side effects related to STH, especially as the majority of workers were illiterate, lacking the basic hygiene knowledge and practices. We recommend the official authority in Jeddah and around the kingdom of Saudi Arabia to revise the standard screening tests for newly arrived workers and enforce regular follow-up inspections to minimize the chances of the spread of STH from expatriate workers to the public.

Keywords: expatriate labors, Jeddah, prevalence, soil transmitted helminths

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63 Possible Involvement of DNA-methyltransferase and Histone Deacetylase in the Regulation of Virulence Potential of Acanthamoeba castellanii

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

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

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

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62 International Coffee Trade in Solidarity with the Zapatista Rebellion: Anthropological Perspectives on Commercial Ethics within Political Antagonistic Movements

Authors: Miria Gambardella

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The influence of solidarity demonstrations towards the Zapatista National Liberation Army has been constantly present over the years, both locally and internationally, guaranteeing visibility to the cause, shaping the movement’s choices, and influencing its hopes of impact worldwide. Most of the coffee produced by the autonomous cooperatives from Chiapas is exported, therefore making coffee trade the main income from international solidarity networks. The question arises about the implications of the relations established between the communities in resistance in Southeastern Mexico and international solidarity movements, specifically on the strategies adopted to conciliate army's demands for autonomy and economic asymmetries between Zapatista cooperatives producing coffee and European collectives who hold purchasing power. In order to deepen the inquiry on those topics, a year-long multi-site investigation was carried out. The first six months of fieldwork were based in Barcelona, where Zapatista coffee was first traded in Spain and where one of the historical and most important European solidarity groups can be found. The last six months of fieldwork were carried out directly in Chiapas, in contact with coffee producers, Zapatista political authorities, international activists as well as vendors, and the rest of the network implicated in coffee production, roasting, and sale. The investigation was based on qualitative research methods, including participatory observation, focus groups, and semi-structured interviews. The analysis did not only focus on retracing the steps of the market chain as if it could be considered a linear and unilateral process, but it rather aimed at exploring actors’ reciprocal perceptions, roles, and dynamics of power. Demonstrations of solidarity and the money circulation they imply aim at changing the system in place and building alternatives, among other things, on the economic level. This work analyzes the formulation of discourse and the organization of solidarity activities that aim at building opportunities for action within a highly politicized economic sphere to which access must be regularly legitimized. The meaning conveyed by coffee is constructed on a symbolic level by the attribution of moral criteria to transactions. The latter participate in the construction of imaginaries that circulate through solidarity movements with the Zapatista rebellion. Commercial exchanges linked to solidarity networks turned out to represent much more than monetary transactions. The social, cultural, and political spheres are invested by ethics, which penetrates all aspects of militant action. It is at this level that the boundaries of different collective actors connect, contaminating each other: merely following the money flow would have been limiting in order to account for a reality within which imaginary is one of the main currencies. The notions of “trust”, “dignity” and “reciprocity” are repeatedly mobilized to negotiate discontinuous and multidirectional flows in the attempt to balance and justify commercial relations in a politicized context that characterizes its own identity through demonizing “market economy” and its dehumanizing powers.

Keywords: coffee trade, economic anthropology, international cooperation, Zapatista National Liberation Army

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61 Functional Plasma-Spray Ceramic Coatings for Corrosion Protection of RAFM Steels in Fusion Energy Systems

Authors: Chen Jiang, Eric Jordan, Maurice Gell, Balakrishnan Nair

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Nuclear fusion, one of the most promising options for reliably generating large amounts of carbon-free energy in the future, has seen a plethora of ground-breaking technological advances in recent years. An efficient and durable “breeding blanket”, needed to ensure a reactor’s self-sufficiency by maintaining the optimal coolant temperature as well as by minimizing radiation dosage behind the blanket, still remains a technological challenge for the various reactor designs for commercial fusion power plants. A relatively new dual-coolant lead-lithium (DCLL) breeder design has exhibited great potential for high-temperature (>700oC), high-thermal-efficiency (>40%) fusion reactor operation. However, the structural material, namely reduced activation ferritic-martensitic (RAFM) steel, is not chemically stable in contact with molten Pb-17%Li coolant. Thus, to utilize this new promising reactor design, the demand for effective corrosion-resistant coatings on RAFM steels represents a pressing need. Solution Spray Technologies LLC (SST) is developing a double-layer ceramic coating design to address the corrosion protection of RAFM steels, using a novel solution and solution/suspension plasma spray technology through a US Department of Energy-funded project. Plasma spray is a coating deposition method widely used in many energy applications. Novel derivatives of the conventional powder plasma spray process, known as the solution-precursor and solution/suspension-hybrid plasma spray process, are powerful methods to fabricate thin, dense ceramic coatings with complex compositions necessary for the corrosion protection in DCLL breeders. These processes can be used to produce ultra-fine molten splats and to allow fine adjustment of coating chemistry. Thin, dense ceramic coatings with chosen chemistry for superior chemical stability in molten Pb-Li, low activation properties, and good radiation tolerance, is ideal for corrosion-protection of RAFM steels. A key challenge is to accommodate its CTE mismatch with the RAFM substrate through the selection and incorporation of appropriate bond layers, thus allowing for enhanced coating durability and robustness. Systematic process optimization is being used to define the optimal plasma spray conditions for both the topcoat and bond-layer, and X-ray diffraction and SEM-EDS are applied to successfully validate the chemistry and phase composition of the coatings. The plasma-sprayed double-layer corrosion resistant coatings were also deposited onto simulated RAFM steel substrates, which are being tested separately under thermal cycling, high-temperature moist air oxidation as well as molten Pb-Li capsule corrosion conditions. Results from this testing on coated samples, and comparisons with bare RAFM reference samples will be presented and conclusions will be presented assessing the viability of the new ceramic coatings to be viable corrosion prevention systems for DCLL breeders in commercial nuclear fusion reactors.

Keywords: breeding blanket, corrosion protection, coating, plasma spray

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60 Covid -19 Pandemic and Impact on Public Spaces of Tourism and Hospitality in Dubai- an Exploratory Study from a Design Perspective

Authors: Manju Bala Jassi

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The Covid 19 pandemic has badly mauled Dubai’s GDP heavily dependent on hospitality, tourism, entertainment, logistics, property and the retail sectors. In the context of the World Health protocols on social distancing for better maintenance of health and hygiene, the revival of the battered tourism and hospitality sectors has serious lessons for designers- interiors and public places. The tangible and intangible aesthetic elements and design –ambiance, materials, furnishings, colors, lighting and interior with architectural design issues of tourism and hospitality need a rethink to ensure a memorable tourist experience. Designers ought to experiment with sustainable places of tourism and design, develop, build and projects are aesthetic and leave as little negative impacts on the environment and public as possible. In short, they ought to conceive public spaces that makes use of little untouched materials and energy, and creates pollution and waste that are minimal. The spaces can employ healthier and more resource-efficient prototypes of construction, renovation, operation, maintenance, and demolition and thereby mitigate the environment impacts of the construction activities and it is sustainable These measures encompass the hospitality sector that includes hotels and restaurants which has taken the hardest fall from the pandemic. The paper sought to examine building energy efficiency and materials and design employed in public places, green buildings to achieve constructive sustainability and to establish the benefits of utilizing energy efficiency, green materials and sustainable design; to document diverse policy interventions, design and Spatial dimensions of tourism and hospitality sectors; to examine changes in the hospitality, aviation sector especially from a design perspective regarding infrastructure or operational constraints and additional risk-mitigation measures; to dilate on the nature of implications for interior designers and architects to design public places to facilitate sustainable tourism and hospitality while balancing convenient space and their operations' natural surroundings. The qualitative research approach was adopted for the study. The researcher collected and analyzed data in continuous iteration. Secondary data was collected from articles in journals, trade publications, government reports, newspaper/ magazine articles, policy documents etc. In depth interviews were conducted with diverse stakeholders. Preliminary data indicates that designers have started imagining public places of tourism and hospitality against the backdrop of the government push and WHO guidelines. For instance, with regard to health, safety, hygiene and sanitation, Emirates, the Dubai-based airline has augmented health measures at the Dubai International Airport and on board its aircraft. It has leveraged high tech/ Nano-tech, social distancing to encourage least human contact, flexible design layouts to limit the occupancy. The researcher organized the data into thematic categories and found that the Government of Dubai has initiated comprehensive measures in the hospitality, tourism and aviation sectors in compliance with the WHO guidelines.

Keywords: Covid 19, design, Dubai, hospitality, public spaces, tourism

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59 Digital Holographic Interferometric Microscopy for the Testing of Micro-Optics

Authors: Varun Kumar, Chandra Shakher

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Micro-optical components such as microlenses and microlens array have numerous engineering and industrial applications for collimation of laser diodes, imaging devices for sensor system (CCD/CMOS, document copier machines etc.), for making beam homogeneous for high power lasers, a critical component in Shack-Hartmann sensor, fiber optic coupling and optical switching in communication technology. Also micro-optical components have become an alternative for applications where miniaturization, reduction of alignment and packaging cost are necessary. The compliance with high-quality standards in the manufacturing of micro-optical components is a precondition to be compatible on worldwide markets. Therefore, high demands are put on quality assurance. For quality assurance of these lenses, an economical measurement technique is needed. For cost and time reason, technique should be fast, simple (for production reason), and robust with high resolution. The technique should provide non contact, non-invasive and full field information about the shape of micro- optical component under test. The interferometric techniques are noncontact type and non invasive and provide full field information about the shape of the optical components. The conventional interferometric technique such as holographic interferometry or Mach-Zehnder interferometry is available for characterization of micro-lenses. However, these techniques need more experimental efforts and are also time consuming. Digital holography (DH) overcomes the above described problems. Digital holographic microscopy (DHM) allows one to extract both the amplitude and phase information of a wavefront transmitted through the transparent object (microlens or microlens array) from a single recorded digital hologram by using numerical methods. Also one can reconstruct the complex object wavefront at different depths due to numerical reconstruction. Digital holography provides axial resolution in nanometer range while lateral resolution is limited by diffraction and the size of the sensor. In this paper, Mach-Zehnder based digital holographic interferometric microscope (DHIM) system is used for the testing of transparent microlenses. The advantage of using the DHIM is that the distortions due to aberrations in the optical system are avoided by the interferometric comparison of reconstructed phase with and without the object (microlens array). In the experiment, first a digital hologram is recorded in the absence of sample (microlens array) as a reference hologram. Second hologram is recorded in the presence of microlens array. The presence of transparent microlens array will induce a phase change in the transmitted laser light. Complex amplitude of object wavefront in presence and absence of microlens array is reconstructed by using Fresnel reconstruction method. From the reconstructed complex amplitude, one can evaluate the phase of object wave in presence and absence of microlens array. Phase difference between the two states of object wave will provide the information about the optical path length change due to the shape of the microlens. By the knowledge of the value of the refractive index of microlens array material and air, the surface profile of microlens array is evaluated. The Sag of microlens and radius of curvature of microlens are evaluated and reported. The sag of microlens agrees well within the experimental limit as provided in the specification by the manufacturer.

Keywords: micro-optics, microlens array, phase map, digital holographic interferometric microscopy

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58 Influence of Water Physicochemical Properties and Vegetation Type on the Distribution of Schistosomiasis Intermediate Host Snails in Nelson Mandela Bay

Authors: Prince S. Campbell, Janine B. Adams, Melusi Thwala, Opeoluwa Oyedele, Paula E. Melariri

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Schistosomiasis is an infectious water-borne disease that holds substantial medical and veterinary importance and is transmitted by Schistosoma flatworms. The transmission and spread of the disease are geographically and temporally confined to water bodies (rivers, lakes, lagoons, dams, etc.) inhabited by its obligate intermediate host snails and human water contact. Human infection with the parasite occurs via skin penetration subsequent to exposure to water infested with schistosome cercariae. Environmental factors play a crucial role in the spread of the disease, as the survival of intermediate host snails is dependent on favourable conditions. These factors include physical and chemical components of water, including pH, salinity, temperature, electrical conductivity, dissolved oxygen, turbidity, water hardness, total dissolved solids, and velocity, as well as biological factors such as predator-prey interactions, competition, food availability, and the presence and density of aquatic vegetation. This study evaluated the physicochemical properties of the water bodies, vegetation type, distribution, and habitat presence of the snail intermediate host. A quantitative cross-sectional research design approach was employed in this study. Eight sampling sites were selected based on their proximity to residential areas. Snails and water physicochemical properties were collected over different seasons for 9 months. A simple dip method was used for surface water samples and measurements were done using multiparameter meters. Snails captured using a 300 µm mesh scoop net and predominant plant species were gathered and transported to experts for identification. Vegetation composition and cover were visually estimated and recorded at each sampling point. Data was analysed using R software (version 4.3.1). A total of 844 freshwater snails were collected, with Physa genera accounting for 95.9% of the snails. Bulinus and Biomphalaria snails, which serve as intermediate hosts for the disease, accounted for (0.9%) and (0.6%) respectively. Indicator macrophytes such as Eicchornia crassipes, Stuckenia pectinate, Typha capensis, and floating macroalgae were found in several water bodies. A negative and weak correlation existed between the number of snails and physicochemical properties such as electrical conductivity (r=-0.240), dissolved oxygen (r=-0.185), hardness (r=-0.210), pH (r=-0.235), salinity (r=-0.242), temperature (r=-0.273), and total dissolved solids (r=-0.236). There was no correlation between the number of snails and turbidity (r=-0.070). Moreover, there was a negative and weak correlation between snails and vegetation coverage (r=-0.127). Findings indicated that snail abundance marginally declined with rising physicochemical concentrations, and the majority of snails were located in regions with less vegetation cover. The reduction in Bulinus and Biomphalaria snail populations may also be attributed to other factors, such as competition among the snails. Snails of the Physa genus were abundant due to their noteworthy resilience in difficult environments. These snails have the potential to function as biological control agents in areas where the disease is endemic, as they outcompete other snails, including schistosomiasis intermediate host snails.

Keywords: intermediate host snails, physicochemical properties, schistosomiasis, vegetation type

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57 3D Printing of Polycaprolactone Scaffold with Multiscale Porosity Via Incorporation of Sacrificial Sucrose Particles

Authors: Mikaela Kutrolli, Noah S. Pereira, Vanessa Scanlon, Mohamadmahdi Samandari, Ali Tamayol

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Bone tissue engineering has drawn significant attention and various biomaterials have been tested. Polymers such as polycaprolactone (PCL) offer excellent biocompatibility, reasonable mechanical properties, and biodegradability. However, PCL scaffolds suffer a critical drawback: a lack of micro/mesoporosity, affecting cell attachment, tissue integration, and mineralization. It also results in a slow degradation rate. While 3D-printing has addressed the issue of macroporosity through CAD-guided fabrication, PCL scaffolds still exhibit poor smaller-scale porosity. To overcome this, we generated composites of PCL, hydroxyapatite (HA), and powdered sucrose (PS). The latter serves as a sacrificial material to generate porous particles after sucrose dissolution. Additionally, we have incorporated dexamethasone (DEX) to boost the PCL osteogenic properties. The resulting scaffolds maintain controlled macroporosity from the lattice print structure but also develop micro/mesoporosity within PCL fibers when exposed to aqueous environments. The study involved mixing PS into solvent-dissolved PCL in different weight ratios of PS to PCL (70:30, 50:50, and 30:70 wt%). The resulting composite was used for 3D printing of scaffolds at room temperature. Printability was optimized by adjusting pressure, speed, and layer height through filament collapse and fusion test. Enzymatic degradation, porogen leaching, and DEX release profiles were characterized. Physical properties were assessed using wettability, SEM, and micro-CT to quantify the porosity (percentage, pore size, and interconnectivity). Raman spectroscopy was used to verify the absence of sugar after leaching. Mechanical characteristics were evaluated via compression testing before and after porogen leaching. Bone marrow stromal cells (BMSCs) behavior in the printed scaffolds was studied by assessing viability, metabolic activity, osteo-differentiation, and mineralization. The scaffolds with a 70% sugar concentration exhibited superior printability and reached the highest porosity of 80%, but performed poorly during mechanical testing. A 50% PS concentration demonstrated a 70% porosity, with an average pore size of 25 µm, favoring cell attachment. No trace of sucrose was found in Raman after leaching the sugar for 8 hours. Water contact angle results show improved hydrophilicity as the sugar concentration increased, making the scaffolds more conductive to cell adhesion. The behavior of bone marrow stromal cells (BMSCs) showed positive viability and proliferation results with an increasing trend of mineralization and osteo-differentiation as the sucrose concentration increased. The addition of HA and DEX also promoted mineralization and osteo-differentiation in the cultures. The integration of PS as porogen at a concentration of 50%wt within PCL scaffolds presents a promising approach to address the poor cell attachment and tissue integration issues of PCL in bone tissue engineering. The method allows for the fabrication of scaffolds with tunable porosity and mechanical properties, suitable for various applications. The addition of HA and DEX further enhanced the scaffolds. Future studies will apply the scaffolds in an in-vivo model to thoroughly investigate their performance.

Keywords: bone, PCL, 3D printing, tissue engineering

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56 Livelihood Security and Mitigating Climate Changes in the Barind Tract of Bangladesh through Agroforestry Systems

Authors: Md Shafiqul Bari, Md Shafiqul Islam Sikdar

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

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

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55 The Istrian Istrovenetian-Croatian Bilingual Corpus

Authors: Nada Poropat Jeletic, Gordana Hrzica

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Bilingual conversational corpora represent a meaningful and the most comprehensive data source for investigating the genuine contact phenomena in non-monitored bi-lingual speech productions. They can be particularly useful for bilingual research since some features of bilingual interaction can hardly be accessed with more traditional methodologies (e.g., elicitation tasks). The method of language sampling provides the resources for describing language interaction in a bilingual community and/or in bilingual situations (e.g. code-switching, amount of languages used, number of languages used, etc.). To capture these phenomena in genuine communication situations, such sampling should be as close as possible to spontaneous communication. Bilingual spoken corpus design is methodologically demanding. Therefore this paper aims at describing the methodological challenges that apply to the corpus design of the conversational corpus design of the Istrian Istrovenetian-Croatian Bilingual Corpus. Croatian is the first official language of the Croatian-Italian officially bilingual Istria County, while Istrovenetian is a diatopic subvariety of Venetian, a longlasting lingua franca in the Istrian peninsula, the mother tongue of the members of the Italian National Community in Istria and the primary code of informal everyday communication among the Istrian Italophone population. Within the CLARIN infrastructure, TalkBank is being used, as it provides relevant procedures for designing and analyzing bilingual corpora. Furthermore, it allows public availability allows for easy replication of studies and cumulative progress as a research community builds up around the corpus, while the tools developed within the field of corpus linguistics enable easy retrieval and analysis of information. The method of language sampling employed is kept at the level of spontaneous communication, in order to maximise the naturalness of the collected conversational data. All speakers have provided written informed consent in which they agree to be recorded at a random point within the period of one month after signing the consent. Participants are administered a background questionnaire providing information about the socioeconomic status and the exposure and language usage in the participants social networks. Recording data are being transcribed, phonologically adapted within a standard-sized orthographic form, coded and segmented (speech streams are being segmented into communication units based on syntactic criteria) and are being marked following the CHAT transcription system and its associated CLAN suite of programmes within the TalkBank toolkit. The corpus consists of transcribed sound recordings of 36 bilingual speakers, while the target is to publish the whole corpus by the end of 2020, by sampling spontaneous conversations among approximately 100 speakers from all the bilingual areas of Istria for ensuring representativeness (the participants are being recruited across three generations of native bilingual speakers in all the bilingual areas of the peninsula). Conversational corpora are still rare in TalkBank, so the Corpus will contribute to BilingBank as a highly relevant and scientifically reliable resource for an internationally established and active research community. The impact of the research of communities with societal bilingualism will contribute to the growing body of research on bilingualism and multilingualism, especially regarding topics of language dominance, language attrition and loss, interference and code-switching etc.

Keywords: conversational corpora, bilingual corpora, code-switching, language sampling, corpus design methodology

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54 Fostering Non-Traditional Student Success in an Online Music Appreciation Course

Authors: Linda Fellag, Arlene Caney

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E-learning has earned an essential place in academia because it promotes learner autonomy, student engagement, and technological aptitude, and allows for flexible learning. However, despite advantages, educators have been slower to embrace e-learning for ESL and other non-traditional students for fear that such students will not succeed without the direct faculty contact and academic support of face-to-face classrooms. This study aims to determine if a non-traditional student-friendly online course can produce student retention and performance rates that compare favorably with those of students in standard online sections of the same course aimed at traditional college-level students. One Music faculty member is currently collaborating with an English instructor to redesign an online college-level Music Appreciation course for non-traditional college students. At Community College of Philadelphia, Introduction to Music Appreciation was recently designated as one of the few college-level courses that advanced ESL, and developmental English students can take while completing their language studies. Beginning in Fall 2017, the course will be critical for international students who must maintain full-time student status under visa requirements. In its current online format, however, Music Appreciation is designed for traditional college students, and faculty who teach these sections have been reluctant to revise the course to address the needs of non-traditional students. Interestingly, presenters maintain that the online platform is the ideal place to develop language and college readiness skills in at-risk students while maintaining the course's curricular integrity. The two faculty presenters describe how curriculum rather than technology drives the redesign of the digitized music course, and self-study materials, guided assignments, and periodic assessments promote independent learning and comprehension of material. The 'scaffolded' modules allow ESL and developmental English students to build on prior knowledge, preview key vocabulary, discuss content, and complete graded tasks that demonstrate comprehension. Activities and assignments, in turn, enhance college success by allowing students to practice academic reading strategies, writing, speaking, and student-faculty and peer-peer communication and collaboration. The course components facilitate a comparison of student performance and retention in sections of the redesigned and existing online sections of Music Appreciation as well as in previous sections with at-risk students. Indirect, qualitative measures include student attitudinal surveys and evaluations. Direct, quantitative measures include withdrawal rates, tests of disciplinary knowledge, and final grades. The study will compare the outcomes of three cohorts in the two versions of the online course: ESL students, at-risk developmental students, and college-level students. These data will also be compared with retention and student outcomes data of the three cohorts in f2f Music Appreciation, which permitted non-traditional student enrollment from 1998-2005. During this eight-year period, the presenter addressed the problems of at-risk students by adding language and college success support, which resulted in strong retention and outcomes. The presenters contend that the redesigned course will produce favorable outcomes among all three cohorts because it contains components which proved successful with at-risk learners in f2f sections of the course. Results of their study will be published in 2019 after the redesigned online course has met for two semesters.

Keywords: college readiness, e-learning, music appreciation, online courses

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53 Absorptive Capabilities in the Development of Biopharmaceutical Industry: The Case of Bioprocess Development and Research Unit, National Polytechnic Institute

Authors: Ana L. Sánchez Regla, Igor A. Rivera González, María del Pilar Monserrat Pérez Hernández

Abstract:

The ability of an organization to identify and get useful information from external sources, assimilate it, transform and apply to generate products or services with added value is called absorptive capacity. Absorptive capabilities contribute to have market opportunities to firms and get a leader position with respect to others competitors. The Bioprocess Development and Research Unit (UDIBI) is a Research and Development (R&D) laboratory that belongs to the National Polytechnic Institute (IPN), which is a higher education institute in Mexico. The UDIBI was created with the purpose of carrying out R and D activities for the Transferon®, a biopharmaceutical product developed and patented by IPN. The evolution of competence and scientific and technological platform made UDIBI expand its scope by providing technological services (preclínical studies and bio-compatibility evaluation) to the national pharmaceutical industry and biopharmaceutical industry. The relevance of this study is that those industries are classified as high scientific and technological intensity, and yet, after a review of the state of the art, there is only one study of absorption capabilities in biopharmaceutical industry with a similar scope to this research; in the case of Mexico, there is none. In addition to this, UDIBI belongs to a public university and its operation does not depend on the federal budget, but on the income generated by its external technological services. This fact represents a highly remarkable case in Mexico's public higher education context. This current doctoral research (2015-2019) is contextualized within a case study, its main objective is to identify and analyze the absorptive capabilities that characterise the UDIBI that allows it had become in a one of two third authorized laboratory by the sanitary authority in Mexico for developed bio-comparability studies to bio-pharmaceutical products. The development of this work in the field is divided into two phases. In a first phase, 15 interviews were conducted with the UDIBI personnel, covering management levels, heads of services, project leaders and laboratory personnel. These interviews were structured under a questionnaire, which was designed to integrate open questions and to a lesser extent, others, whose answers would be answered on a Likert-type rating scale. From the information obtained in this phase, a scientific article was made (in review and a proposal of presentation was submitted in different academic forums. A second stage will be made from the conduct of an ethnographic study within this organization under study that will last about 3 months. On the other hand, it is intended to carry out interviews with external actors around the UDIBI (suppliers, advisors, IPN officials, including contact with an academic specialized in absorption capacities to express their comments on this thesis. The inicial findings had shown two lines: i) exist institutional, technological and organizational management elements that encourage and/or limit the creation of absorption capacities in this scientific and technological laboratory and, ii) UDIBI has had created a set of multiple transfer technology of knowledge mechanisms which have had permitted to build a huge base of prior knowledge.

Keywords: absorptive capabilities, biopharmaceutical industry, high research and development intensity industries, knowledge management, transfer of knowledge

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52 Superhydrophobic Materials: A Promising Way to Enhance Resilience of Electric System

Authors: M. Balordi, G. Santucci de Magistris, F. Pini, P. Marcacci

Abstract:

The increasing of extreme meteorological events represents the most important causes of damages and blackouts of the whole electric system. In particular, the icing on ground-wires and overheads lines, due to snowstorms or harsh winter conditions, very often gives rise to the collapse of cables and towers both in cold and warm climates. On the other hand, the high concentration of contaminants in the air, due to natural and/or antropic causes, is reflected in high levels of pollutants layered on glass and ceramic insulators, causing frequent and unpredictable flashover events. Overheads line and insulator failures lead to blackouts, dangerous and expensive maintenances and serious inefficiencies in the distribution service. Inducing superhydrophobic (SHP) properties to conductors, ground-wires and insulators, is one of the ways to face all these problems. Indeed, in some cases, the SHP surface can delay the ice nucleation time and decrease the ice nucleation temperature, preventing ice formation. Besides, thanks to the low surface energy, the adhesion force between ice and a superhydrophobic material are low and the ice can be easily detached from the surface. Moreover, it is well known that superhydrophobic surfaces can have self-cleaning properties: these hinder the deposition of pollution and decrease the probability of flashover phenomena. Here this study presents three different studies to impart superhydrophobicity to aluminum, zinc and glass specimens, which represent the main constituent materials of conductors, ground-wires and insulators, respectively. The route to impart the superhydrophobicity to the metallic surfaces can be summarized in a three-step process: 1) sandblasting treatment, 2) chemical-hydrothermal treatment and 3) coating deposition. The first step is required to create a micro-roughness. In the chemical-hydrothermal treatment a nano-scale metallic oxide (Al or Zn) is grown and, together with the sandblasting treatment, bring about a hierarchical micro-nano structure. By coating an alchilated or fluorinated siloxane coating, the surface energy decreases and gives rise to superhydrophobic surfaces. In order to functionalize the glass, different superhydrophobic powders, obtained by a sol-gel synthesis, were prepared. Further, the specimens were covered with a commercial primer and the powders were deposed on them. All the resulting metallic and glass surfaces showed a noticeable superhydrophobic behavior with a very high water contact angles (>150°) and a very low roll-off angles (<5°). The three optimized processes are fast, cheap and safe, and can be easily replicated on industrial scales. The anti-icing and self-cleaning properties of the surfaces were assessed with several indoor lab-tests that evidenced remarkable anti-icing properties and self-cleaning behavior with respect to the bare materials. Finally, to evaluate the anti-snow properties of the samples, some SHP specimens were exposed under real snow-fall events in the RSE outdoor test-facility located in Vinadio, western Alps: the coated samples delay the formation of the snow-sleeves and facilitate the detachment of the snow. The good results for both indoor and outdoor tests make these materials promising for further development in large scale applications.

Keywords: superhydrophobic coatings, anti-icing, self-cleaning, anti-snow, overheads lines

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51 Stent Surface Functionalisation via Plasma Treatment to Promote Fast Endothelialisation

Authors: Irene Carmagnola, Valeria Chiono, Sandra Pacharra, Jochen Salber, Sean McMahon, Chris Lovell, Pooja Basnett, Barbara Lukasiewicz, Ipsita Roy, Xiang Zhang, Gianluca Ciardelli

Abstract:

Thrombosis and restenosis after stenting procedure can be prevented by promoting fast stent wall endothelialisation. It is well known that surface functionalisation with antifouling molecules combining with extracellular matrix proteins is a promising strategy to design biomimetic surfaces able to promote fast endothelialization. In particular, REDV has gained much attention for the ability to enhance rapid endothelialization due to its specific affinity with endothelial cells (ECs). In this work, a two-step plasma treatment was performed to polymerize a thin layer of acrylic acid, used to subsequently graft PEGylated-REDV and polyethylene glycol (PEG) at different molar ratio with the aim to selectively promote endothelial cell adhesion avoiding platelet activation. PEGylate-REDV was provided by Biomatik and it is formed by 6 PEG monomer repetitions (Chempep Inc.), with an NH2 terminal group. PEG polymers were purchased from Chempep Inc. with two different chain lengths: m-PEG6-NH2 (295.4 Da) with 6 monomer repetitions and m-PEG12-NH2 (559.7 Da) with 12 monomer repetitions. Plasma activation was obtained by operating at 50W power, 5 min of treatment and at an Ar flow rate of 20 sccm. Pure acrylic acid (99%, AAc) vapors were diluted in Ar (flow = 20 sccm) and polymerized by a pulsed plasma discharge applying a discharge RF power of 200 W, a duty cycle of 10% (on time = 10 ms, off time = 90 ms) for 10 min. After plasma treatment, samples were dipped into an 1-(3-dimethylaminopropyl)-3- ethylcarbodiimide (EDC)/N-hydroxysuccinimide (NHS) solution (ratio 4:1, pH 5.5) for 1 h at 4°C and subsequently dipped in PEGylate-REDV and PEGylate-REDV:PEG solutions at different molar ratio (100 μg/mL in PBS) for 20 h at room temperature. Surface modification was characterized through physico-chemical analyses and in vitro cell tests. PEGylated-REDV peptide and PEG were successfully bound to the carboxylic groups that are formed on the polymer surface after plasma reaction. FTIR-ATR spectroscopy, X -ray Photoelectron Spectroscopy (XPS) and contact angle measurement gave a clear indication of the presence of the grafted molecules. The use of PEG as a spacer allowed for an increase in wettability of the surface, and the effect was more evident by increasing the amount of PEG. Endothelial cells adhered and spread well on the surfaces functionalized with the REDV sequence. In conclusion, a selective coating able to promote a new endothelial cell layer on polymeric stent surface was developed. In particular, a thin AAc film was polymerised on the polymeric surface in order to expose –COOH groups, and PEGylate-REDV and PEG were successful grafted on the polymeric substrates. The REDV peptide demonstrated to encourage cell adhesion with a consequent, expected improvement of the hemocompatibility of these polymeric surfaces in vivo. Acknowledgements— This work was funded by the European Commission 7th Framework Programme under grant agreement number 604251- ReBioStent (Reinforced Bioresorbable Biomaterials for Therapeutic Drug Eluting Stents). The authors thank all the ReBioStent partners for their support in this work.

Keywords: endothelialisation, plasma treatment, stent, surface functionalisation

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50 Wear Resistance in Dry and Lubricated Conditions of Hard-anodized EN AW-4006 Aluminum Alloy

Authors: C. Soffritti, A. Fortini, E. Baroni, M. Merlin, G. L. Garagnani

Abstract:

Aluminum alloys are widely used in many engineering applications due to their advantages such ashigh electrical and thermal conductivities, low density, high strength to weight ratio, and good corrosion resistance. However, their low hardness and poor tribological properties still limit their use in industrial fields requiring sliding contacts. Hard anodizing is one of the most common solution for overcoming issues concerning the insufficient friction resistance of aluminum alloys. In this work, the tribological behavior ofhard-anodized AW-4006 aluminum alloys in dry and lubricated conditions was evaluated. Three different hard-anodizing treatments were selected: a conventional one (HA) and two innovative golden hard-anodizing treatments (named G and GP, respectively), which involve the sealing of the porosity of anodic aluminum oxides (AAO) with silver ions at different temperatures. Before wear tests, all AAO layers were characterized by scanning electron microscopy (VPSEM/EDS), X-ray diffractometry, roughness (Ra and Rz), microhardness (HV0.01), nanoindentation, and scratch tests. Wear tests were carried out according to the ASTM G99-17 standard using a ball-on-disc tribometer. The tests were performed in triplicate under a 2 Hz constant frequency oscillatory motion, a maximum linear speed of 0.1 m/s, normal loads of 5, 10, and 15 N, and a sliding distance of 200 m. A 100Cr6 steel ball10 mm in diameter was used as counterpart material. All tests were conducted at room temperature, in dry and lubricated conditions. Considering the more recent regulations about the environmental hazard, four bio-lubricants were considered after assessing their chemical composition (in terms of Unsaturation Number, UN) and viscosity: olive, peanut, sunflower, and soybean oils. The friction coefficient was provided by the equipment. The wear rate of anodized surfaces was evaluated by measuring the cross-section area of the wear track with a non-contact 3D profilometer. Each area value, obtained as an average of four measurements of cross-section areas along the track, was used to determine the wear volume. The worn surfaces were analyzed by VPSEM/EDS. Finally, in agreement with DoE methodology, a statistical analysis was carried out to identify the most influencing factors on the friction coefficients and wear rates. In all conditions, results show that the friction coefficient increased with raising the normal load. Considering the wear tests in dry sliding conditions, irrespective of the type of anodizing treatments, metal transfer between the mating materials was observed over the anodic aluminum oxides. During sliding at higher loads, the detachment of the metallic film also caused the delamination of some regions of the wear track. For the wear tests in lubricated conditions, the natural oils with high percentages of oleic acid (i.e., olive and peanut oils) maintained high friction coefficients and low wear rates. Irrespective of the type of oil, smallmicrocraks were visible over the AAO layers. Based on the statistical analysis, the type of anodizing treatment and magnitude of applied load were the main factors of influence on the friction coefficient and wear rate values. Nevertheless, an interaction between bio-lubricants and load magnitude could occur during the tests.

Keywords: hard anodizing treatment, silver ions, bio-lubricants, sliding wear, statistical analysis

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49 Design of Experiment for Optimizing Immunoassay Microarray Printing

Authors: Alex J. Summers, Jasmine P. Devadhasan, Douglas Montgomery, Brittany Fischer, Jian Gu, Frederic Zenhausern

Abstract:

Immunoassays have been utilized for several applications, including the detection of pathogens. Our laboratory is in the development of a tier 1 biothreat panel utilizing Vertical Flow Assay (VFA) technology for simultaneous detection of pathogens and toxins. One method of manufacturing VFA membranes is with non-contact piezoelectric dispensing, which provides advantages, such as low-volume and rapid dispensing without compromising the structural integrity of antibody or substrate. Challenges of this processinclude premature discontinuation of dispensing and misaligned spotting. Preliminary data revealed the Yp 11C7 mAb (11C7)reagent to exhibit a large angle of failure during printing which may have contributed to variable printing outputs. A Design of Experiment (DOE) was executed using this reagent to investigate the effects of hydrostatic pressure and reagent concentration on microarray printing outputs. A Nano-plotter 2.1 (GeSIM, Germany) was used for printing antibody reagents ontonitrocellulose membrane sheets in a clean room environment. A spotting plan was executed using Spot-Front-End software to dispense volumes of 11C7 reagent (20-50 droplets; 1.5-5 mg/mL) in a 6-test spot array at 50 target membrane locations. Hydrostatic pressure was controlled by raising the Pressure Compensation Vessel (PCV) above or lowering it below our current working level. It was hypothesized that raising or lowering the PCV 6 inches would be sufficient to cause either liquid accumulation at the tip or discontinue droplet formation. After aspirating 11C7 reagent, we tested this hypothesis under stroboscope.75% of the effective raised PCV height and of our hypothesized lowered PCV height were used. Humidity (55%) was maintained using an Airwin BO-CT1 humidifier. The number and quality of membranes was assessed after staining printed membranes with dye. The droplet angle of failure was recorded before and after printing to determine a “stroboscope score” for each run. The DOE set was analyzed using JMP software. Hydrostatic pressure and reagent concentration had a significant effect on the number of membranes output. As hydrostatic pressure was increased by raising the PCV 3.75 inches or decreased by lowering the PCV -4.5 inches, membrane output decreased. However, with the hydrostatic pressure closest to equilibrium, our current working level, membrane output, reached the 50-membrane target. As the reagent concentration increased from 1.5 to 5 mg/mL, the membrane output also increased. Reagent concentration likely effected the number of membrane output due to the associated dispensing volume needed to saturate the membranes. However, only hydrostatic pressure had a significant effect on stroboscope score, which could be due to discontinuation of dispensing, and thus the stroboscope check could not find a droplet to record. Our JMP predictive model had a high degree of agreement with our observed results. The JMP model predicted that dispensing the highest concentration of 11C7 at our current PCV working level would yield the highest number of quality membranes, which correlated with our results. Acknowledgements: This work was supported by the Chemical Biological Technologies Directorate (Contract # HDTRA1-16-C-0026) and the Advanced Technology International (Contract # MCDC-18-04-09-002) from the Department of Defense Chemical and Biological Defense program through the Defense Threat Reduction Agency (DTRA).

Keywords: immunoassay, microarray, design of experiment, piezoelectric dispensing

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48 Isolation of Bacterial Species with Potential Capacity for Siloxane Removal in Biogas Upgrading

Authors: Ellana Boada, Eric Santos-Clotas, Alba Cabrera-Codony, Maria Martin, Lluis Baneras, Frederic Gich

Abstract:

Volatile methylsiloxanes (VMS) are a group of manmade silicone compounds widely used in household and industrial applications that end up on the biogas produced through the anaerobic digestion of organic matter in landfills and wastewater treatment plants. The presence of VMS during the biogas energy conversion can cause damage on the engines, reducing the efficiency of this renewable energy source. Non regenerative adsorption onto activated carbon is the most widely used technology to remove siloxanes from biogas, while new trends point out that biotechnology offers a low-cost and environmentally friendly alternative to conventional technologies. The first objective of this research was to enrich, isolate and identify bacterial species able to grow using siloxane molecules as a sole carbon source: anoxic wastewater sludge was used as initial inoculum in liquid anoxic enrichments, adding D4 (as representative siloxane compound) previously adsorbed on activated carbon. After several months of acclimatization, liquid enrichments were plated onto solid media containing D4 and thirty-four bacterial isolates were obtained. 16S rRNA gene sequencing allowed the identification of strains belonging to the following species: Ciceribacter lividus, Alicycliphilus denitrificans, Pseudomonas aeruginosa and Pseudomonas citronellolis which are described to be capable to degrade toxic volatile organic compounds. Kinetic assays with 8 representative strains revealed higher cell growth in the presence of D4 compared to the control. Our second objective was to characterize the community composition and diversity of the microbial community present in the enrichments and to elucidate whether the isolated strains were representative members of the community or not. DNA samples were extracted, the 16S rRNA gene was amplified (515F & 806R primer pair), and the microbiome analyzed from sequences obtained with a MiSeq PE250 platform. Results showed that the retrieved isolates only represented a minor fraction of the microorganisms present in the enrichment samples, which were represented by Alpha, Beta, and Gamma proteobacteria as dominant groups in the category class thus suggesting that other microbial species and/or consortia may be important for D4 biodegradation. These results highlight the need of additional protocols for the isolation of relevant D4 degraders. Currently, we are developing molecular tools targeting key genes involved in siloxane biodegradation to identify and quantify the capacity of the isolates to metabolize D4 in batch cultures supplied with a synthetic gas stream of air containing 60 mg m⁻³ of D4 together with other volatile organic compounds found in the biogas mixture (i.e. toluene, hexane and limonene). The isolates were used as inoculum in a biotrickling filter containing lava rocks and activated carbon to assess their capacity for siloxane removal. Preliminary results of biotrickling filter performance showed 35% of siloxane biodegradation in a contact time of 14 minutes, denoting that biological siloxane removal is a promising technology for biogas upgrading.

Keywords: bacterial cultivation, biogas upgrading, microbiome, siloxanes

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47 The Vanishing Treasure: An Anthropological Study on Changing Social Relationships, Values, Belief System and Language Pattern of the Limbus in Kalimpong Sub-Division of the Darjeeling District in West Bengal, India

Authors: Biva Samadder, Samita Manna

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India is a melting pot of races, tribes, castes and communities. The population of India can be roughly branched into the huge majority of “Civilized” Indians of the Plains and the minority of Tribal population of the hill area and the forest who constituting almost 16 percent of total population of India. The Kirat community composed of four ethnic tribes: Limbu, Lepcha, Dhimal, and Rai. These Kirat people were found to be rich in indigenous knowledge, skill and practices especially for the use on medicinal plants and livelihood purposes. The “Mundhum" is the oral scripture or the “Bible of the Limbus” which serves as the canon of the codes of the Limbu socialization, their moral values and the very orientation of their lifestyle. From birth till death the Limbus are disciplined in the life with full of religious rituals, traditions and culture governed by community norms with a rich legacy of indigenous knowledge and traditional practices. The present study has been conducted using both secondary as well as primary data by applying social methodology consisting of the social survey, questionnaire, interviews and observations in the Kalimpong Block-I of Darjeeling District of west Bengal of India, which is a heterogeneous zone in terms of its ethnic composition and where the Limbus are pre-dominantly concentrated. Due to their close contact with other caste and communities Limbus are now adjusted with the changing situation by borrowing some cultural traits from the other communities and changes that have taken place in their cultural practices, religious beliefs, economic aspects, languages and in social roles and relationships which is bringing the change in their material culture. Limbu language is placed in the Tibeto- Burman Language category. But due to the political and cultural domination of educationally sound and numerically dominant Bengali race, the different communities in this area forced to come under the one umbrella of the Nepali or Gorkhali nation (nation-people). Their respective identities had to be submerged in order to constitute as a strong force to resist Nepali domination and ensure their common survival. As Nepali is a lingua-franca of the area knowing and speaking Nepali language helps them in procuring economic and occupational facilities. Ironically, present day younger generation does not feel comfortable speaking in their own Limbu tongue. The traditional knowledge about medicinal plants, healing, and health culture is found to be wear away due to the lack of interest of young generation. Not only poverty, along with exclusion due to policies they are in the phase of extinction, but their capabilities are ignored and not documented and preserved especially in the case of Limbus who having a great cultural heritage of an oral tradition. Attempts have been made to discuss the persistence and changes in socioeconomic pattern of life in relation to the social structure, material culture, cultural practices, social relationships, indigenous technology, ethos and their values and belief system.

Keywords: changing social relationship, cultural transition, identity, indigenous knowledge, language

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46 Differential Survival Rates of Pseudomonas aeruginosa Strains on the Wings of Pantala flavescens

Authors: Banu Pradheepa Kamarajan, Muthusamy Ananthasubramanian

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Biofilm forming Pseudomonads occupy the top third position in causing hospital acquired infections. P. aeruginosa is notoriously known for its tendency to develop drug resistance. Major classes of drug such as β-lactams, aminoglycosides, quinolones, and polymyxins are found ineffective against multi-drug resistance Pseudomonas. To combat the infections, rather than administration of a single antibiotic, use of combinations (tobramycin and essential oils from plants and/or silver nanoparticles, chitosan, nitric oxide, cis-2-decenoic acid) in single formulation are suggested to control P. aeruginosa biofilms. Conventional techniques to prevent hospital-acquired implant infections such as coatings with antibiotics, controlled release of antibiotics from the implant material, contact-killing surfaces, coating the implants with functional DNase I and, coating with glycoside hydrolase are being followed. Coatings with bioactive components besides having limited shelf-life, require cold-chain and, are likely to fail when bacteria develop resistance. Recently identified nano-scale physical architectures on the insect wings are expected to have potential bactericidal property. Nanopillars are bactericidal to Staphylococcus aureus, Bacillus subtilis, K. pnuemoniae and few species of Pseudomonas. Our study aims to investigate the survival rate of biofilm forming Pseudomonas aeruginosa strain over non-biofilm forming strain on the nanopillar architecture of dragonfly (Pantala flavescens) wing. Dragonflies were collected near house-hold areas and, insect identification was carried out by the Department of Entomology, Tamilnadu Agricultural University, Coimbatore, India. Two strains of P. aeruginosa such as PAO1 (potent biofilm former) and MTCC 1688 (non-weak biofilm former) were tested against the glass coverslip (control) and wings of dragonfly (test) for 48 h. The wings/glass coverslips were incubated with bacterial suspension in 48-well plate. The plates were incubated at 37 °C under static condition. Bacterial attachment on the nanopillar architecture of the wing surface was visualized using FESEM. The survival rate of P. aeruginosa was tested using colony counting technique and flow cytometry at 0.5 h, 1 h, 2 h, 7 h, 24 h, and 48 h post-incubation. Cell death was analyzed using propidium iodide staining and DNA quantification. The results indicated that the survival rate of non-biofilm forming P. aeruginosa is 0.2 %, whilst that of biofilm former is 45 % on the dragonfly wings at the end of 48 h. The reduction in the survival rate of biofilm and non-biofilm forming P. aeruginosa was 20% and 40% respectively on the wings compared to the glass coverslip. In addition, Fourier Transformed Infrared Radiation was used to study the modification in the surface chemical composition of the wing during bacterial attachment and, post-sonication. This result indicated that the chemical moieties are not involved in the bactericidal property of nanopillars by the conserved characteristic peaks of chitin pre and post-sonication. The nanopillar architecture of the dragonfly wing efficiently deters the survival of non-biofilm forming P. aeruginosa, but not the biofilm forming strain. The study highlights the ability of biofilm formers to survive on wing architecture. Understanding this survival strategy will help in designing the architecture that combats the colonization of biofilm forming pathogens.

Keywords: biofilm, nanopillars, Pseudomonas aeruginosa, survival rate

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