Search results for: laser scattering

Authors: J. Gabrielczyk, J. Kluitmann, T. Dammeyer, H. J. Jördening

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High-level expression of proteins in bacteria often causes production of insoluble protein aggregates, called inclusion bodies (IB). They contain mainly one type of protein and offer an easy and efficient way to get purified protein. On the other hand, proteins in IB are normally devoid of function and therefore need a special treatment to become active. Most refolding techniques aim at diluting the solubilizing chaotropic agents. Unfortunately, optimal refolding conditions have to be found empirically for every protein. For large-scale applications, a simple refolding process with high yields and high final enzyme concentrations is still missing. The constructed plasmid pASK-IBA63b containing the sequence of fructosyltransferase (FTF, EC 2.4.1.162) from Bacillus subtilis NCIMB 11871 was transformed into E. coli BL21 (DE3) Rosetta. The bacterium was cultivated in a fed-batch bioreactor. The produced FTF was obtained mainly as IB. For refolding experiments, five different amounts of IBs were solubilized in urea buffer with protein concentration of 0.2-8.5 g/L. Solubilizates were refolded with batch or continuous dialysis. The refolding yield was determined by measuring the protein concentration of the clear supernatant before and after the dialysis. Particle size was measured by dynamic light scattering. We tested the solubilization properties of fructosyltransferase IBs. The particle size measurements revealed that the solubilization of the aggregates is achieved at urea concentration of 5M or higher and confirmed by absorption spectroscopy. All results confirm previous investigations that refolding yields are dependent upon initial protein concentration. In batch dialysis, the yields dropped from 67% to 12% and 72% to 19% for continuous dialysis, in relation to initial concentrations from 0.2 to 8.5 g/L. Often used additives such as sucrose and glycerol had no effect on refolding yields. Buffer screening indicated a significant increase in activity but also temperature stability of FTF with citrate/phosphate buffer. By adding citrate to the dialysis buffer, we were able to increase the refolding yields to 82-47% in batch and 90-74% in the continuous process. Further experiments showed that in general, higher ionic strength of buffers had major impact on refolding yields; doubling the buffer concentration increased the yields up to threefold. Finally, we achieved corresponding high refolding yields by reducing the chamber volume by 75% and the amount of buffer needed. The refolded enzyme had an optimal activity of 12.5±0.3 x104 units/g. However, detailed experiments with native FTF revealed a reaggregation of the molecules and loss in specific activity depending on the enzyme concentration and particle size. For that reason, we actually focus on developing a process of simultaneous enzyme refolding and immobilization. The results of this study show a new approach in finding optimal refolding conditions for inclusion bodies at high concentrations. Straightforward buffer screening and increase of the ionic strength can optimize the refolding yield of the target protein by 400%. Gentle removal of chaotrope with continuous dialysis increases the yields by an additional 65%, independent of the refolding buffer applied. In general time is the crucial parameter for successful refolding of solubilized proteins.

Keywords: dialysis, inclusion body, refolding, solubilization

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Authors: Lea Boidin, Martha Baydoun, Bertrand Leroux, Olivier Morales, Samir Acherar, Celine Frochot, Nadira Delhem

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Ovarian cancer (OC) is one of the most defying diseases in gynecologic oncology. Even though surgery remains crucial in the therapy of patients with primary ovarian cancer, recurrent recidivism calls for the development of new therapy protocols to propose for patients dealing with this cancer. FRα is described as a tumor‐associated antigen in OC, where FRα expression is usually linked with more poorly differentiated, aggressive tumors. The Photodynamic treatment (PDT) available data have shown improvements in the uptake of small tumors and in the induction of a proper anti-tumoral immune response. In order to target specifically peritoneal metastatis, which overexpress FRα, a new-patented PS coupled with folic acid has been developed in our team. Herein we propose PDT using this new patented PS for PDT applied in an in vivo mice model. The efficacy of the treatment was evaluated in mice without and with PBMC reconstitution. Mice were divided into four groups: Non-Treated, PS, Light Only, and PDT Treated and subjected to illumination by laser set at 668nm with a duration of illumination of 45 minutes (or 1 min of illumination followed by 2 minutes of pause repeated 45 times). When mice were not reconstituted and after fractionized PDT protocol, a significant decrease in the tumor volume was noticed. An induction in the anti-tumoral cytokine IFNγ chaperoned this decrease while a subsequent inhibition in the cytokine TGFβ. Even more crucial, when mice were reconstituted and upon PDT, the fold of tumor decrease was even higher. An immune response was activated decoded with an increase in NK, CD3 +, LT helper and Cytotoxic T cells. Thereafter, an increase in the expression of the cytokines IFNγ and TNFα were noticed while an inhibition in TGFβ, IL8 and IL10 accompanied this immune response activation. Therefore, our work has shown for the first time that a fractionized PDT protocol using a folate-targeted PDT is effective for treatment of ovarian cancer. The interest in using PDT in this case, goes beyond the local induction of tumor apoptosis only, but can promote subsequent anti-tumor response. Most of the therapies currently used to treat ovarian cancer, have an uncooperative outcomes on the host immune response. The readiness of a tumor adjuvant treatment like PDT adequate in eliminating the tumor and in concert stimulating anti-tumor immunity would be weighty.

Keywords: folate receptor, ovarian cancer, photodynamic therapy, humanized mice model

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Authors: V. Ziegler, F. Schneider, M. Pesch

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With the EDM264, GRIMM offers a solution for mobile short- and long-term measurements in outdoor areas and at production sites. For research as well as permanent areal observations on a near reference quality base. The model EDM264 features a powerful and robust measuring cell based on optical particle counting (OPC) principle with all the advantages that users of GRIMM's portable aerosol spectrometers are used to. The system is embedded in a compact weather-protection housing with all-weather sampling, heated inlet system, data logger, and meteorological sensor. With TSP, PM10, PM4, PM2.5, PM1, and PMcoarse, the EDM264 provides all fine dust fractions real-time, valid for outdoor applications and calculated with the proven GRIMM enviro-algorithm, as well as six additional dust mass fractions pm10, pm2.5, pm1, inhalable, thoracic and respirable for IAQ and workplace measurements. This highly versatile instrument performs real-time monitoring of particle number, particle size and provides information on particle surface distribution as well as dust mass distribution. GRIMM's EDM264 has 31 equidistant size channels, which are PSL traceable. A high-end data logger enables data acquisition and wireless communication via LTE, WLAN, or wired via Ethernet. Backup copies of the measurement data are stored in the device directly. The rinsing air function, which protects the laser and detector in the optical cell, further increases the reliability and long term stability of the EDM264 under different environmental and climatic conditions. The entire sample volume flow of 1.2 L/min is analyzed by 100% in the optical cell, which assures excellent counting efficiency at low and high concentrations and complies with the ISO 21501-1standard for OPCs. With all these features, the EDM264 is a world-leading dust monitor for precise monitoring of particulate matter and particle number concentration. This highly reliable instrument is an indispensable tool for many users who need to measure aerosol levels and air quality outdoors, on construction sites, or at production facilities.

Keywords: aerosol research, aerial observation, fence line monitoring, wild fire detection

Procedia PDF Downloads 151

Authors: Natalia Lukasik, Ewa Wagner-Wysiecka

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Development of selective, fast-responsive, and economical sensors for diverse ions detection and determination is one of the most extensively studied areas due to its importance in the field of clinical, environmental and industrial analysis. Among chemical sensors, vast popularity has gained ionophore-based optical sensors, where the generated analytical signal is a consequence of the molecular recognition of ion by the ionophore. Change of color occurring during host-guest interactions allows for quantitative analysis and for 'naked-eye' detection without the need of using sophisticated equipment. An example of application of such sensors is colorimetric detection of iron(III) cations. Iron as one of the most significant trace elements plays roles in many biochemical processes. For these reasons, the development of reliable, fast, and selective methods of iron ions determination is highly demanded. Taking all mentioned above into account a chromogenic amide derivative of 3,4-dihydroxybenzoic acid was synthesized, and its ability to iron(III) recognition was tested. To the best of authors knowledge (according to chemical abstracts) the obtained ligand has not been described in the literature so far. The catechol moiety was introduced to the ligand structure in order to mimic the action of naturally occurring siderophores-iron(III)-selective receptors. The ligand–ion interactions were studied using spectroscopic methods: UV-Vis spectrophotometry and infrared spectroscopy. The spectrophotometric measurements revealed that the amide exhibits affinity to iron(III) in dimethyl sulfoxide and fully aqueous solution, what is manifested by the change of color from yellow to green. Incorporation of the tested amide into a polymeric matrix (cellulose triacetate) ensured effective recognition of iron(III) at pH 3 with the detection limit 1.58×10⁻⁵ M. For the obtained sensor material parameters like linear response range, response time, selectivity, and possibility of regeneration were determined. In order to evaluate the effect of the size of the sensing material on iron(III) detection nanospheres (in the form of nanoemulsion) containing the tested amide were also prepared. According to DLS (dynamic light scattering) measurements, the size of the nanospheres is 308.02 ± 0.67 nm. Work parameters of the nanospheres were determined and compared with cellulose triacetate-based material. Additionally, for fast, qualitative experiments the test strips were prepared by adsorption of the amide solution on a glass microfiber material. Visual limit of detection of iron(III) at pH 3 by the test strips was estimated at the level 10⁻⁴ M. In conclusion, reported here amide derived from 3,4- dihydroxybenzoic acid proved to be an effective candidate for optical sensing of iron(III) in fully aqueous solutions. N. L. kindly acknowledges financial support from National Science Centre Poland the grant no. 2017/01/X/ST4/01680. Authors thank for financial support from Gdansk University of Technology grant no. 032406.

Keywords: ion-selective optode, iron(III) recognition, nanospheres, optical sensor

Procedia PDF Downloads 154

Authors: Iva Blazkova, Amitava Moulick, Vedran Milosavljevic, Pavel Kopel, Marketa Vaculovicova, Vojtech Adam, Rene Kizek

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Doxorubicin is one of the most commonly used and the most effective chemotherapeutic drugs. This antracycline drug isolated from the bacteria Streptomyces peuceticus var. caesius is sold under the trade name Adriamycin (hydroxydaunomycin, hydroxydaunorubicin). Doxorubicin is used in single therapy to treat hematological malignancies (blood cancers, leukaemia, lymphoma), many types of carcinoma (solid tumors) and soft tissue sarcomas. It has many serious side effects like nausea and vomiting, hair lost, myelosupression, oral mucositis, skin reactions and redness, but the most serious one is the cardiotoxicity. Because of the risk of heart attack and congestive heart failure, the total dose administered to patients has to be accurately monitored. With the aim to lower the side effects and to targeted delivery of doxorubicin into the tumor tissue, the different nanoparticles are studied. The drug can be bound on a surface of nanoparticle, encapsulated in the inner cavity, or incorporated into the structure of nanoparticle. Among others, carbon nanoparticles (graphene, carbon nanotubes, fullerenes) are highly studied. Besides the number of inorganic nanoparticles, a great potential exhibit also organic ones mainly lipid-based and polymeric nanoparticle. The aim of this work was to perform a toxicity study of free doxorubicin compared to doxorubicin conjugated with various nanotransporters. The effect of liposomes, fullerenes, graphene, and carbon nanotubes on the toxicity was analyzed. As a first step, the binding efficacy of between doxorubicin and the nanotransporter was determined. The highest efficacy was detected in case of liposomes (85% of applied drug was encapsulated) followed by graphene, carbon nanotubes and fullerenes. For the toxicological studies, the chicken embryos incubated under controlled conditions (37.5 °C, 45% rH, rotation every 2 hours) were used. In 7th developmental day of chicken embryos doxorubicin or doxorubicin-nanotransporter complex was applied on the chorioallantoic membrane of the eggs and the viability was analyzed every day till the 17th developmental day. Then the embryos were extracted from the shell and the distribution of doxorubicin in the body was analyzed by measurement of organs extracts using laser induce fluorescence detection. The chicken embryo mortality caused by free doxorubicin (30%) was significantly lowered by using the conjugation with nanomaterials. The highest accumulation of doxorubicin and doxorubicin nanotransporter complexes was observed in the liver tissue

Keywords: doxorubicin, chicken embryos, nanotransporters, toxicity

Procedia PDF Downloads 449

Authors: Adan Valdez, Shawn Gallaher, James Morison, Jordan Aragon

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Global climate change has impacted atmospheric temperatures contributing to rising sea levels, decreasing sea ice, and increased freshening of high latitude oceans. This freshening has contributed to increased stratification inhibiting local mixing and nutrient transport and modifying regional circulations in polar oceans. In recent years, the Western Arctic has seen an increase in freshwater volume at an average rate of 397+-116 km3/year. The majority of the freshwater volume resides in the Beaufort Gyre surface lens driven by anticyclonic wind forcing, sea ice melt, and Arctic river runoff. The total climatological freshwater content is typically defined as water fresher than 34.8. The near-isothermal nature of Arctic seawater and non-linearities in the equation of state for near-freezing waters result in a salinity driven pycnocline as opposed to the temperature driven density structure seen in the lower latitudes. In this study, we investigate the relationship between freshwater content and remotely sensed dynamic ocean topography (DOT). In-situ measurements of freshwater content are useful in providing information on the freshening rate of the Beaufort Gyre; however, their collection is costly and time consuming. NASA’s Advanced Topographic Laser Altimeter System (ATLAS) derived dynamic ocean topography (DOT), and Air Expendable CTD (AXCTD) derived Freshwater Content are used to develop a linear regression model. In-situ data for the regression model is collected across the 150° West meridian, which typically defines the centerline of the Beaufort Gyre. Two freshwater content models are determined by integrating the freshwater volume between the surface and an isopycnal corresponding to reference salinities of 28.7 and 34.8. These salinities correspond to those of the winter pycnocline and total climatological freshwater content, respectively. Using each model, we determine the strength of the linear relationship between freshwater content and satellite derived DOT. The result of this modeling study could provide a future predictive capability of freshwater volume changes in the Beaufort-Chukchi Sea using non in-situ methods. Successful employment of the ICESat-2’s DOT approximation of freshwater content could potentially reduce reliance on field deployment platforms to characterize physical ocean properties.

Keywords: ICESat-2, dynamic ocean topography, freshwater content, beaufort gyre

Procedia PDF Downloads 87

Authors: Cynthia Briggs, Kimberly Gerardi

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Over the past six years, the authors have been working together on World Language Collaborations in the Middle School French Program at St. Luke's School in New Canaan, Connecticut, USA. Author 2 brings design expertise to the projects, and both teachers have utilized the fabrication lab, emerging technologies, and collaboration with students. Each year, author 1 proposes a project scope, and her students are challenged to design and engineer a signature project. Both partners have improved the iterative process to ensure deeper learning and sustained student inquiry. The projects range from a 1:32 scale model of the Eiffel Tower that was CNC routed to a fully functional jukebox that plays francophone music, lights up, and can hold up to one thousand songs powered by Raspberry Pi. The most recent project is a Fragrance Marketplace, culminating with a pop-up store for the entire community to discover. Each student will learn the history of fragrance and the chemistry behind making essential oils. Students then create a unique brand, marketing strategy, and concept for their signature fragrance. They are further tasked to use the industrial design process (bottling, packaging, and creating a brand name) to finalize their product for the public Marketplace. Sometimes, these dynamic projects require maintenance and updates. For example, our wall-mounted, three-foot francophone clock is constantly changing. The most recent iteration uses Chat GPT to program the Arduino to reconcile the real-time clock shield and keep perfect time as each hour passes. The lights, motors, and sounds from the clock are authentic to each region, represented with laser-cut embellishments. Inspired by Michel Parmigiani, the history of Swiss watch-making, and the precision of time instruments, we aim for perfection with each passing minute. The authors aim to share exemplary work that is possible with students of all ages. We implemented the reverse engineering process to focus on student outcomes to refine our collaborative process. The products that our students create are prime examples of how the design engineering process is applicable across disciplines. The authors firmly believe that the past and present of World cultures inspire innovation.

Keywords: collaboration, design thinking, emerging technologies, world language

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Authors: Sutapa Mondal Roy, Suban K. Sahoo

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The primary aim of this work is to synthesis silver nanoparticles (AgNPs) through environmentally benign routes to avoid any chemical toxicity related undesired side effects. The nanoparticles were stabilized with drug ciprofloxacin (Cp) and were studied for their effectiveness as drug delivery agent. Targeted drug delivery improves the therapeutic potential of drugs at the diseased site as well as lowers the overall dose and undesired side effects. The small size of nanoparticles greatly facilitates the transport of active agents (drugs) across biological membranes and allows them to pass through the smallest capillaries in the body that are 5-6 μm in diameter, and can minimize possible undesired side effects. AgNPs are non-toxic, inert, stable, and has a high binding capacity and thus can be considered as biomaterials. AgNPs were synthesized from the nutrient broth supernatant after the culture of environment-friendly bacteria Bacillus subtilis. The AgNPs were found to show the surface plasmon resonance (SPR) band at 425 nm. The Cp capped Ag nanoparticles formation was complete within 30 minutes, which was confirmed from absorbance spectroscopy. Physico-chemical nature of the AgNPs-Cp system was confirmed by Dynamic Light Scattering (DLS), Transmission Electron Microscopy (TEM) etc. The AgNPs-Cp system size was found to be in the range of 30-40 nm. To monitor the kinetics of drug release from the surface of nanoparticles, the release of Cp was carried out by careful dialysis keeping AgNPs-Cp system inside the dialysis bag at pH 7.4 over time. The drug release was almost complete after 30 hrs. During the drug delivery process, to understand the AgNPs-Cp system in a better way, the sincere theoretical investigation is been performed employing Density Functional Theory. Electronic charge transfer, electron density, binding energy as well as thermodynamic properties like enthalpy, entropy, Gibbs free energy etc. has been predicted. The electronic and thermodynamic properties, governed by the AgNPs-Cp interactions, indicate that the formation of AgNPs-Cp system is exothermic i.e. thermodynamically favorable process. The binding energy and charge transfer analysis implies the optimum stability of the AgNPs-Cp system. Thus, the synthesized Cp-Ag nanoparticles can be effectively used for biological purposes due to its environmentally benign routes of synthesis procedures, which is clean, biocompatible, non-toxic, safe, cost-effective, sustainable and eco-friendly. The Cp-AgNPs as biomaterials can be successfully used for drug delivery procedures due to slow release of drug from nanoparticles over a considerable period of time. The kinetics of the drug release show that this drug-nanoparticle assembly can be effectively used as potential tools for therapeutic applications. The ease of synthetic procedure, lack of possible chemical toxicity and their biological activity along with excellent application as drug delivery agent will open up vista of using nanoparticles as effective and successful drug delivery agent to be used in modern days.

Keywords: silver nanoparticles, ciprofloxacin, density functional theory, drug delivery

Procedia PDF Downloads 384

Authors: Anvesh Gaddam, Amit Agrawal, Suhas Joshi, Mark Thompson

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An increase in surface area to volume ratio with a decrease in characteristic length scale, leads to a rapid increase in pressure drop across the microchannel. Texturing the microchannel surfaces reduce the effective surface area, thereby decreasing the pressured drop. Surface texturing introduces two wetting states: a metastable Cassie-Baxter state and stable Wenzel state. Predicting wetting transition in textured microchannels is essential for identifying optimal parameters leading to maximum drag reduction. Optical methods allow visualization only in confined areas, therefore, obtaining whole-field information on wetting transition is challenging. In this work, we propose a non-invasive method to capture wetting transitions in textured microchannels under flow conditions. To this end, we tracked the behavior of the Poiseuille number Po = f.Re, (with f the friction factor and Re the Reynolds number), for a range of flow rates (5 < Re < 50), and different wetting states were qualitatively demarcated by observing the inflection points in the f.Re curve. Microchannels with both longitudinal and transverse ribs with a fixed gas fraction (δ, a ratio of shear-free area to total area) and at a different confinement ratios (ε, a ratio of rib height to channel height) were fabricated. The measured pressure drop values for all the flow rates across the textured microchannels were converted into Poiseuille number. Transient behavior of the pressure drop across the textured microchannels revealed the collapse of liquid-gas interface into the gas cavities. Three wetting states were observed at ε = 0.65 for both longitudinal and transverse ribs, whereas, an early transition occurred at Re ~ 35 for longitudinal ribs at ε = 0.5, due to spontaneous flooding of the gas cavities as the liquid-gas interface ruptured at the inlet. In addition, the pressure drop in the Wenzel state was found to be less than the Cassie-Baxter state. Three-dimensional numerical simulations confirmed the initiation of the completely wetted Wenzel state in the textured microchannels. Furthermore, laser confocal microscopy was employed to identify the location of the liquid-gas interface in the Cassie-Baxter state. In conclusion, the present method can overcome the limitations posed by existing techniques, to conveniently capture wetting transition in textured microchannels.

Keywords: drag reduction, Poiseuille number, textured surfaces, wetting transition

Procedia PDF Downloads 161

Authors: Elisabeth Unterfrauner, Christian Voigt

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Since the foundation of the first Fab Lab by the Massachusetts Institute of Technology about 17 years ago, the Maker movement has spread globally with the foundation of maker spaces and Fab Labs worldwide. In these workshops, citizens have access to digital fabrication technologies such as 3D printers and laser cutters to develop and test their own ideas and prototypes, which makes it an attractive place for start-up companies. Know-How is shared not only in the physical space but also online in diverse communities. According to the Horizon report, the Maker movement, however, will also have an impact on educational settings in the following years. The European project ‘DOIT - Entrepreneurial skills for young social innovators in an open digital world’ has incorporated key elements of making to develop an early entrepreneurial education program for children between the age of six and 16. The Maker pedagogy builds on constructive learning approaches, learning by doing principles, learning in collaborative and interdisciplinary teams and learning through trial and error where mistakes are acknowledged as learning opportunities. The DOIT program consists of seven consecutive elements. It starts with a motivation phase where students get motivated by envisioning the scope of their possibilities. The second step is about Co-design: Students are asked to collect and select potential ideas for innovations. In the Co-creation phase students gather in teams and develop first prototypes of their ideas. In the iteration phase, the prototype is continuously improved and in the next step, in the reflection phase, feedback on the prototypes is exchanged between the teams. In the last two steps, scaling and reaching out, the robustness of the prototype is tested with a bigger group of users outside of the educational setting and finally students will share their projects with a wider public. The DOIT program involves 1,000 children in two pilot phases at 11 pilot sites in ten different European countries. The comprehensive evaluation design is based on a mixed method approach with a theoretical backbone on Lackeus’ model of entrepreneurship education, which distinguishes between entrepreneurial attitudes, entrepreneurial skills and entrepreneurial knowledge. A pre-post-test with quantitative measures as well as qualitative data from interviews with facilitators, students and workshop protocols will reveal the effectiveness of the program. The evaluation results will be presented at the conference.

Keywords: early entrepreneurial education, Fab Lab, maker education, Maker movement

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Authors: Francisco Pereira, António Augusto Vicente, Filipe Vaz, Joel Borges, Pedro Geada

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Light is a growth-limiting factor in microalgae cultivation, where factors like spectral components, intensity, and duration, often characterized by its wavelength, are well-reported to have a substantial impact on cell growth rates and, consequently, photosynthetic performance and mitigation of CO2, one of the most significant greenhouse gases (GHGs). Photobioreactors (PBRs) are commonly used to grow microalgae under controlled conditions, but they often fail to provide an even light distribution to the cultures. For this reason, there is a pressing need for innovations aiming at enhancing the efficient utilization of light. So, one potential approach to address this issue is by implementing plasmonic films, such as the localized surface plasmon resonance (LSPR). LSPR is an optical phenomenon connected to the interaction of light with metallic nanostructures. LSPR excitation is characterized by the oscillation of unbound conduction electrons of the nanoparticles coupled with the electromagnetic field from incident light. As a result of this excitation, highly energetic electrons and a strong electromagnetic field are generated. These effects lead to an amplification of light scattering, absorption, and extinction of specific wavelengths, contingent on the nature of the employed nanoparticle. Thus, microalgae might benefit from this biotechnology as it enables the selective filtration of inhibitory wavelengths and harnesses the electromagnetic fields produced, which could lead to enhancements in both biomass and metabolite productivity. This study aimed at implementing and evaluating a “plasmon-enhanced PBR”. The goal was to utilize LSPR thin films to enhance the growth and CO2 bio-fixation rate of Chlorella vulgaris. The internal/external walls of the PBRs were coated with a TiO2 matrix containing different nanoparticles (Au, Ag, and Au-Ag) in order to evaluate the impact of this approach on microalgae’s performance. Plasmonic films with distinct compositions resulted in different Chlorella vulgaris growth, ranging from 4.85 to 6.13 g.L-1. The highest cell concentrations were obtained with the metallic Ag films, demonstrating a 14% increase compared to the control condition. Moreover, it appeared to be no differences in growth between PBRs with inner and outer wall coatings. In terms of CO2 bio-fixation, distinct rates were obtained depending on the coating applied, ranging from 0.42 to 0.53 gCO2L-1d-1. Ag coating was demonstrated to be the most effective condition for carbon fixation by C. vulgaris. The impact of LSPR films on the biochemical characteristics of biomass (e.g., proteins, lipids, pigments) was analysed as well. Interestingly, Au coating yielded the most significant enhancements in protein content and total pigments, with increments of 15 % and 173 %, respectively, when compared to the PBR without any coating (control condition). Overall, the incorporation of plasmonic films in PBRs seems to have the potential to improve the performance and efficiency of microalgae cultivation, thereby representing an interesting approach to increase both biomass production and GHGs bio-mitigation.

Keywords: CO₂ bio-fixation, plasmonic effect, photobioreactor, photosynthetic microalgae

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Authors: A. S. V. Carvalho, J. Luis, L. S. O. Pires, J. M. Oliveira

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Additive manufacturing technologies (AM) experienced a remarkable growth in the latest years due to the development and diffusion of a wide range of three-dimensional (3D) printing techniques. Nowadays we can find techniques available for non-industrial users, like fused filament fabrication, but techniques like 3D printing, polyjet, selective laser sintering and stereolithography are mainly spread in the industry. Robocasting (R3D) shows a great potential due to its ability to shape materials with a wide range of viscosity. Industrial porcelain compositions showing different rheological behaviour can be prepared and used as candidate materials to be processed by R3D. The use of this AM technique in industry is very residual. In this work, a specific porcelain composition with suitable rheological properties will be processed by R3D, and a systematic study of the printing parameters tuning will be shown. The porcelain composition was formulated based on an industrial spray dried porcelain powder. The powder particle size and morphology was analysed. The powders were mixed with water and an organic binder on a ball mill at 200 rpm/min for 24 hours. The batch viscosity was adjusted by the addition of an acid solution and mixed again. The paste density, viscosity, zeta potential, particle size distribution and pH were determined. In a R3D system, different speed and pressure settings were studied to access their impact on the fabrication of porcelain models. These models were dried at 80 °C, during 24 hours and sintered in air at 1350 °C for 2 hours. The stability of the models, its walls and surface quality were studied and their physical properties were accessed. The microstructure and layer adhesion were observed by SEM. The studied processing parameters have a high impact on the models quality. Moreover, they have a high impact on the stacking of the filaments. The adequate tuning of the parameters has a huge influence on the final properties of the porcelain models. This work contributes to a better assimilation of AM technologies in ceramic industry. Acknowledgments: The RoboCer3D project – project of additive rapid manufacturing through 3D printing ceramic material (POCI-01-0247-FEDER-003350) financed by Compete 2020, PT 2020, European Regional Development Fund – FEDER through the International and Competitive Operational Program (POCI) under the PT2020 partnership agreement.

Keywords: additive manufacturing, porcelain, robocasting, R3D

Procedia PDF Downloads 162

Authors: Nino Kupatadze, Mzevinar Bedinashvili, Tamar Memanishvili, Manana Gurielidze, David Tugushi, Ramaz Katsarava

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Bacteria easily colonize the surfaces of tissues, surgical devices (implants, orthopedics, catheters, etc.), and instruments causing surgical device related infections. Therefore, the battle against bacteria and the prevention of surgical devices from biofilm formation is one of the main challenges of biomedicine today. Our strategy to the solution of this problem consists in using antimicrobial polymeric coatings as effective “shields” to protect surfaces from bacteria’s colonization and biofilm formation. As one of the most promising approaches look be the use of antimicrobial bioerodible polymeric nanocomposites containing silver nanoparticles (AgNPs). We assume that the combination of an erodible polymer with a strong bactericide should put obstacles to bacteria to occupy the surface and to form biofilm. It has to be noted that this kind of nanocomposites are also promising as wound dressing materials to treat infected superficial wounds. Various synthetic and natural polymers were used for creating biocomposites containing AgNPs as both particles' stabilizers and matrices forming elastic films at surfaces. One of the most effective systems to fabricate AgNPs is an ethanol solution of polyvinylpyrrolidone(PVP) with dissolved AgNO3–ethanol serves as a AgNO3 reductant and PVP as AgNPs stabilizer (through the interaction of nanoparticles with nitrogen atom of the amide group). Though PVP is biocompatible and film-forming polymer, it is not a good candidate to design either "biofilm shield" or wound dressing material because of a high solubility in water – though the solubility of PVP provides the desirable release of AgNPs from the matrix, but the coating is easily washable away from the surfaces. More promising as matrices look water insoluble but bioerodible polymers that can provide the release of AgNPs and form long-lasting coatings at the surfaces. For creating bioerodible water-insoluble antimicrobial coatings containing AgNPs, we selected amino acid based biodegradable polymers(AABBPs)–poly(ester amide)s, poly(ester urea)s, their copolymers containing amide and related groups capable to stabilize AgNPs. Among a huge variety of AABBPs reported we selected the polymers soluble in ethanol. For preparing AgNPs containing nanocompositions AABBPs and AgNO3 were dissolved in ethanol and subjected to photochemical reduction using daylight-irradiation. The formation of AgNPs was observed visually by coloring the solutions in brownish-red. The obtained AgNPs were characterized by UV-spectroscopy, transmission electron microscopy(TEM), and dynamic light scattering(DLS). According to the UV and TEM data, the photochemical reduction resulted presumably in spherical AgNPs with rather high contribution of the particles below 10 nm that are known as responsible for the antimicrobial activity. DLS study showed that average size of nanoparticles formed after photo-reduction in ethanol solution ranged within 50 nm. The in vitro antimicrobial activity study of the new nanocomposite material is in progress now.

Keywords: nanocomposites, silver nanoparticles, polymer, biodegradable

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Authors: Quan Wen, Tianxiao Chang, Shaolu Shi, Yushi Wang, Guangyu Wang

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As a kind of working environment of the fuze, the spin rate decaying law of projectile in exterior trajectory is of great value in the design of the rotation count fixed distance fuze. In addition, it is significant in the field of devices for simulation tests of fuze exterior ballistic environment, flight stability, and dispersion accuracy of gun projectile and opening and scattering design of submunition and illuminating cartridges. Besides, the self-destroying mechanism of the fuze in small-caliber projectile often works by utilizing the attenuation of centrifugal force. In the theory of projectile aerodynamics and fuze design, there are many formulas describing the change law of projectile angular velocity in external ballistic such as Roggla formula, exponential function formula, and power function formula. However, these formulas are mostly semi-empirical due to the poor test conditions and insufficient test data at that time. These formulas are difficult to meet the design requirements of modern fuze because they are not accurate enough and have a narrow range of applications now. In order to provide more accurate ballistic environment parameters for the design of a hemispherical head projectile fuze, the projectile’s spin rate decaying law in exterior trajectory under the effect of air resistance was studied. In the analysis, the projectile shape was simplified as hemisphere head, cylindrical part, rotating band part, and anti-truncated conical tail. The main assumptions are as follows: a) The shape and mass are symmetrical about the longitudinal axis, b) There is a smooth transition between the ball hea, c) The air flow on the outer surface is set as a flat plate flow with the same area as the expanded outer surface of the projectile, and the boundary layer is turbulent, d) The polar damping moment attributed to the wrench hole and rifling mark on the projectile is not considered, e) The groove of the rifle on the rotating band is uniform, smooth and regular. The impacts of the four parts on aerodynamic moment of the projectile rotation were obtained by aerodynamic theory. The surface friction stress of the projectile, the polar damping moment formed by the head of the projectile, the surface friction moment formed by the cylindrical part, the rotating band, and the anti-truncated conical tail were obtained by mathematical derivation. After that, the mathematical model of angular spin rate attenuation was established. In the whole trajectory with the maximum range angle (38°), the absolute error of the polar damping torque coefficient obtained by simulation and the coefficient calculated by the mathematical model established in this paper is not more than 7%. Therefore, the credibility of the mathematical model was verified. The mathematical model can be described as a first-order nonlinear differential equation, which has no analytical solution. The solution can be only gained as a numerical solution by connecting the model with projectile mass motion equations in exterior ballistics.

Keywords: ammunition engineering, fuze technology, spin rate, numerical simulation

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Authors: Astrid Tannert, Anuradha Ramoji, Christina Ebert, Frederike Gladigau, Lorena Tuchscherr, Jürgen Popp, Ute Neugebauer

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Staphylococcus aureus is a facultative intracellular pathogen, which by entering host cells may evade immunologic host response as well as antimicrobial treatment. In that way, S. aureus can cause persistent intracellular infections which are difficult to treat. Depending on the strain, S. aureus may persist at different intracellular locations like the phagolysosome. The first barrier invading pathogens from the blood stream that they have to cross are the endothelial cells lining the inner surface of blood and lymphatic vessels. Upon proceeding from an acute to a chronic infection, intracellular pathogens undergo certain biochemical and structural changes including a deceleration of metabolic processes to adopt for long-term intracellular survival and the development of a special phenotype designated as small colony variant. In this study, the endothelial cell line Ea.hy 926 was used as a model for acute and chronic S. aureus infection. To this end, Ea.hy 926 cells were cultured on QIAscout™ Microraft Arrays, a special graded cell culture substrate that contains around 12,000 microrafts of 200 µm edge length. After attachment to the substrate, the endothelial cells were infected with GFP-expressing S. aureus for 3 weeks. The acute infection and the development of persistent bacteria was followed by confocal laser scanning microscopy, scanning the whole Microraft Array for the presence and for detailed determination of the intracellular location of fluorescent intracellular bacteria every second day. After three weeks of infection representative microrafts containing infected cells, cells with protruded infections and cells that did never show any infection were isolated and fixed for Raman micro-spectroscopic investigation. For comparison, also microrafts with acute infection were isolated. The acquired Raman spectra are correlated with the fluorescence microscopic images to give hints about a) the molecular alterations in endothelial cells during acute and chronic infection compared to non-infected cells, and b) metabolic and structural changes within the pathogen when entering a mode of persistence within host cells. We thank Dr. Ruth Kläver from QIAGEN GmbH for her support regarding QIAscout technology. Financial support by the BMBF via the CSCC (FKZ 01EO1502) and from the DFG via the Jena Biophotonic and Imaging Laboratory (JBIL, FKZ PO 633/29-1, BA 1601/10-1) is highly acknowledged.

Keywords: correlative image analysis, intracellular infection, pathogen-host adaption, Raman micro-spectroscopy

Procedia PDF Downloads 181

Authors: Istvan Szilagyi, Marko Pavlovic, Paul Rouster

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Antioxidant enzymes are the most efficient defense systems against reactive oxygen species, which cause severe damage in living organisms and industrial products. However, their supplementation is problematic due to their high sensitivity to the environmental conditions. Immobilization on carrier nanoparticles is a promising research direction towards the improvement of their functional and colloidal stability. In that way, their applications in biomedical treatments and manufacturing processes in the food, textile and cosmetic industry can be extended. The main goal of the present research was to prepare and formulate antioxidant bionanocomposites composed of superoxide dismutase (SOD) enzyme, anionic clay (layered double hydroxide, LDH) nanoparticle and heparin (HEP) polyelectrolyte. To characterize the structure and the colloidal stability of the obtained compounds in suspension and solid state, electrophoresis, dynamic light scattering, transmission electron microscopy, spectrophotometry, thermogravimetry, X-ray diffraction, infrared and fluorescence spectroscopy were used as experimental techniques. LDH-SOD composite was synthesized by enzyme immobilization on the clay particles via electrostatic and hydrophobic interactions, which resulted in a strong adsorption of the SOD on the LDH surface, i.e., no enzyme leakage was observed once the material was suspended in aqueous solutions. However, the LDH-SOD showed only limited resistance against salt-induced aggregation and large irregularly shaped clusters formed during short term interval even at lower ionic strengths. Since sufficiently high colloidal stability is a key requirement in most of the applications mentioned above, the nanocomposite was coated with HEP polyelectrolyte to develop highly stable suspensions of primary LDH-SOD-HEP particles. HEP is a natural anticoagulant with one of the highest negative line charge density among the known macromolecules. The experimental results indicated that it strongly adsorbed on the oppositely charged LDH-SOD surface leading to charge inversion and to the formation of negatively charged LDH-SOD-HEP. The obtained hybrid materials formed stable suspension even under extreme conditions, where classical colloid chemistry theories predict rapid aggregation of the particles and unstable suspensions. Such a stabilization effect originated from electrostatic repulsion between the particles of the same sign of charge as well as from steric repulsion due to the osmotic pressure raised during the overlap of the polyelectrolyte chains adsorbed on the surface. In addition, the SOD enzyme kept its structural and functional integrity during the immobilization and coating processes and hence, the LDH-SOD-HEP bionanocomposite possessed excellent activity in decomposition of superoxide radical anions, as revealed in biochemical test reactions. In conclusion, due to the improved colloidal stability and the good efficiency in scavenging superoxide radical ions, the developed enzymatic system is a promising antioxidant candidate for biomedical or other manufacturing processes, wherever the aim is to decompose reactive oxygen species in suspensions.

Keywords: clay, enzyme, polyelectrolyte, formulation

Procedia PDF Downloads 268

Authors: Nadine Raidl, Benjamin Kaufmann, Michael Hofstätter, Peter Supancic

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If polycrystalline ZnO is properly doped and sintered under very specific conditions, it shows unique electrical properties, which are indispensable for today’s electronic industries, where it is used as the number one overvoltage protection material. Under a critical voltage, the polycrystalline bulk exhibits high electrical resistance but becomes suddenly up to twelve magnitudes more conductive if this voltage limit is exceeded (i.e., varistor effect). It is known that these peerless properties have their origin in the grain boundaries of the material. Electric charge is accumulated in the boundaries, causing a depletion layer in their vicinity and forming potential barriers (so-called Double Schottky Barriers, or DSB) which are responsible for the highly non-linear conductivity. Since ZnO is a piezoelectric material, mechanical stresses induce polarisation charges that modify the DSB heights and as a result the global electrical characteristics (i.e., piezotronic effect). In this work, a finite element method was used to simulate emerging stresses on individual grains in the bulk. Besides, experimental efforts were made to testify a coherent model that could explain this influence. Electron back scattering diffraction was used to identify grain orientations. With the help of wet chemical etching, grain polarization was determined. Micro lock-in infrared thermography (MLIRT) was applied to detect current paths through the material, and a micro 4-point probes method system (M4PPS) was employed to investigate current-voltage characteristics between single grains. Bulk samples were tested under uniaxial pressure. It was found that the conductivity can increase by up to three orders of magnitude with increasing stress. Through in-situ MLIRT, it could be shown that this effect is caused by the activation of additional current paths in the material. Further, compressive tests were performed on miniaturized samples with grain paths containing solely one or two grain boundaries. The tests evinced both an increase of the conductivity, as observed for the bulk, as well as a decreased conductivity. This phenomenon has been predicted theoretically and can be explained by piezotronically induced surface charges that have an impact on the DSB at the grain boundaries. Depending on grain orientation and stress direction, DSB can be raised or lowered. Also, the experiments revealed that the conductivity within one single specimen can increase and decrease, depending on the current direction. This novel finding indicates the existence of asymmetric Double Schottky Barriers, which was furthermore proved by complementary methods. MLIRT studies showed that the intensity of heat generation within individual current paths is dependent on the direction of the stimulating current. M4PPS was used to study the relationship between the I-V characteristics of single grain boundaries and grain orientation and revealed asymmetric behavior for very specific orientation configurations. A new model for the Double Schottky Barrier, taking into account the natural asymmetry and explaining the experimental results, will be given.

Keywords: Asymmetric Double Schottky Barrier, piezotronic, varistor, zinc oxide

Procedia PDF Downloads 267

Authors: L. Serrano-Lara, J.M Alba-Gómez

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Qubbet el–Hawa Project has been documented archaeological artifacts as 3d models by laser scanning technique since 2015. Currently, research has obtained the right methodology to develop a high accuracy photographic texture for each geometrical 3D model. Furthermore, the right methodology to attach the complete digital surrogate into a 3DPDF document has been obtained; it is used as a catalogue worksheet that brings archaeological data and, at the same time, allows us to obtain precise measurements, volume calculations and cross-section mapping of each scanned artifact. This validated archaeological documentation is the first step for dissemination, application as Qubbet el-Hawa Virtual Museum, and, moreover, multi-sensory experience through 3D print archaeological artifacts. Material culture from four funerary complexes constructed in West Aswan has become physical replicas opening the archaeological research process itself and offering creative possibilities on museology or educational projects. This paper shares a method of acquiring texture for scanning´s output product in order to achieve a 3DPDF archaeological cataloguing, and, on the other hand, to allow the colorfully 3D printing of singular archaeological artifacts. The proposed method has undergone two concrete cases, a polychrome wooden ushabti, and, a cartonnage mask belonging to a lady, bought recovered on intact tomb QH34aa. Both 3D model results have been implemented on three main applications, archaeological 3D catalogue, public dissemination activities, and the 3D artifact model in a bachelor education program. Due to those three already mentioned applications, productive interaction among spectator and three-dimensional artifact have been increased; moreover, functionality as archaeological documentation has been consolidated. Finding the right methodology to assign a specific color to each vector on the geometric 3D model, we had been achieved two essential archaeological applications. Firstly, 3DPDF as a display document for an archaeological catalogue, secondly, the possibility to obtain a colored 3d printed object to be displayed in public exhibitions. Obsolescences 3D models have become updated archaeological documentation of QH43aa tomb cultural material. Therefore, Qubbet el-Hawa Project has been actualized the educational potential of its results thanks to a multi-sensory experience that arose from 3d scanned´s archaeological artifacts.

Keywords: 3D printed, 3D scanner, Middle Kingdom, Qubbet el-Hawa necropolis, virtual archaeology

Procedia PDF Downloads 141

Authors: K. Mokatse, O. M. Barei, K. Gabanakgosi, P. Matlhabaphiri

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The mining of pit slopes is often associated with some level of deviation from design recommendations, and this may translate to associated changes in the stability of the excavated pit slopes. Therefore slope angle design reconciliations are essential for assessing and monitoring compliance of excavated pit slopes to accepted slope designs. These associated changes in slope stability may be reflected by changes in the calculated factors of safety and/or probabilities of failure. Reconciliations of as-mined and slope design profiles are conducted periodically to assess the implications of these deviations on pit slope stability. Currently, the slope design reconciliation process being implemented in Jwaneng Mine involves the measurement of as-mined and design slope angles along vertical sections cut along the established geotechnical design section lines on the GEOVIA GEMS™ software. Bench retentions are calculated as a percentage of the available catchment area, less over-mined and under-mined areas, to that of the designed catchment area. This process has proven to be both tedious and requires a lot of manual effort and time to execute. Consequently, a new semi-automated mine-to-design reconciliation approach that utilizes laser scanning and GIS-based tools is being proposed at Jwaneng Mine. This method involves high-resolution scanning of targeted bench walls, subsequent creation of 3D surfaces from point cloud data and the derivation of slope toe lines and crest lines on the Maptek I-Site Studio software. The toe lines and crest lines are then exported to the ArcGIS software where distance offsets between the design and actual bench toe lines and crest lines are calculated. Retained bench catchment capacity is measured as distances between the toe lines and crest lines on the same bench elevations. The assessment of the performance of the inter-ramp and overall slopes entails the measurement of excavated and design slope angles along vertical sections on the ArcGIS software. Excavated and design toe-to-toe or crest-to-crest slope angles are measured for inter-ramp stack slope reconciliations. Crest-to-toe slope angles are also measured for overall slope angle design reconciliations. The proposed approach allows for a more automated, accurate, quick and easier workflow for carrying out slope angle design reconciliations. This process has proved highly effective and timeous in the assessment of slope performance in Jwaneng Mine. This paper presents a newly proposed process for assessing compliance to slope angle designs for Jwaneng Mine.

Keywords: slope angle designs, slope design recommendations, slope performance, slope stability

Procedia PDF Downloads 237

Authors: Rima Zakzouk, Yasushi Shimada, Yasunori Sumi, Junji Tagami

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Background and purpose: The resin composite has become the main material for the restorations of caries in recent years due to aesthetic characteristics, especially with the development of the adhesive techniques. The quality of adhesion to tooth structures is depending on an exchange process between inorganic tooth material and synthetic resin and a micromechanical retention promoted by resin infiltration in partially demineralized dentin. Optical coherence tomography (OCT) is a noninvasive diagnostic method for obtaining cross-sectional images that produce high-resolution of the biological tissue at the micron scale. The aim of this study was to evaluate the gap formation at adhesive/tooth interface of two-step self-etch adhesives that are preceded with or without phosphoric acid pre-etching in different regions of teeth using SS-OCT. Materials and methods: Round tapered cavities (2×2 mm) were prepared in cervical part of bovine incisors teeth and divided into 2 groups (n=10): first group self-etch adhesive (Clearfil SE Bond) was applied for SE group and second group treated with acid etching before applying the self-etch adhesive for PA group. Subsequently, both groups were restored with Estelite Flow Quick Flowable Composite Resin and observed under OCT. Following 5000 thermal cycles, the same section was obtained again for each cavity using OCT at 1310-nm wavelength. Scanning was repeated after two months to monitor the gap progress. Then the gap length was measured using image analysis software, and the statistics analysis were done between both groups using SPSS software. After that, the cavities were sectioned and observed under Confocal Laser Scanning Microscope (CLSM) to confirm the result of OCT. Results: Gaps formed at the bottom of the cavity was longer than the gap formed at the margin and dento-enamel junction in both groups. On the other hand, pre-etching treatment led to damage the DEJ regions creating longer gap. After 2 months the results showed almost progress in the gap length significantly at the bottom regions in both groups. In conclusions, phosphoric acid etching treatment did not reduce the gap lrngth in most regions of the cavity. Significance: The bottom region of tooth was more exposed to gap formation than margin and DEJ regions, The DEJ damaged with phosphoric acid treatment.

Keywords: optical coherence tomography, self-etch adhesives, bottom, dento enamel junction

Procedia PDF Downloads 227

Authors: Tibor Ajtai, Máté Pintér, Noémi Utry, Gergely Kiss-Albert, Andrea Palágyi, László Manczinger, Csaba Vágvölgyi, Gábor Szabó, Zoltán Bozóki

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According to the best knowledge of the authors, here we experimentally demonstrate first, a quantified correlation between the real-time measured optical feature of the ambient and the off-line measured toxicity data. Finally, using these correlations we are presenting a novel methodology for real time characterisation of ambient toxicity based on the multi wavelength aerosol phase photoacoustic measurement. Ambient carbonaceous particulate matter is one of the most intensively studied atmospheric constituent in climate science nowadays. Beyond their climatic impact, atmospheric soot also plays an important role as an air pollutant that harms human health. Moreover, according to the latest scientific assessments ambient soot is the second most important anthropogenic emission source, while in health aspect its being one of the most harmful atmospheric constituents as well. Despite of its importance, generally accepted standard methodology for the quantitative determination of ambient toxicology is not available yet. Dominantly, ambient toxicology measurement is based on the posterior analysis of filter accumulated aerosol with limited time resolution. Most of the toxicological studies are based on operational definitions using different measurement protocols therefore the comprehensive analysis of the existing data set is really limited in many cases. The situation is further complicated by the fact that even during its relatively short residence time the physicochemical features of the aerosol can be masked significantly by the actual ambient factors. Therefore, decreasing the time resolution of the existing methodology and developing real-time methodology for air quality monitoring are really actual issues in the air pollution research. During the last decades many experimental studies have verified that there is a relation between the chemical composition and the absorption feature quantified by Absorption Angström Exponent (AAE) of the carbonaceous particulate matter. Although the scientific community are in the common platform that the PhotoAcoustic Spectroscopy (PAS) is the only methodology that can measure the light absorption by aerosol with accurate and reliable way so far, the multi-wavelength PAS which are able to selectively characterise the wavelength dependency of absorption has become only available in the last decade. In this study, the first results of the intensive measurement campaign focusing the physicochemical and toxicological characterisation of ambient particulate matter are presented. Here we demonstrate the complete microphysical characterisation of winter time urban ambient including optical absorption and scattering as well as size distribution using our recently developed state of the art multi-wavelength photoacoustic instrument (4λ-PAS), integrating nephelometer (Aurora 3000) as well as single mobility particle sizer and optical particle counter (SMPS+C). Beyond this on-line characterisation of the ambient, we also demonstrate the results of the eco-, cyto- and genotoxicity measurements of ambient aerosol based on the posterior analysis of filter accumulated aerosol with 6h time resolution. We demonstrate a diurnal variation of toxicities and AAE data deduced directly from the multi-wavelength absorption measurement results.

Keywords: photoacoustic spectroscopy, absorption Angström exponent, toxicity, Ames-test

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Authors: Tais Basaco, Stefanie Pektor, Josue A. Moreno, Matthias Miederer, Andreas Türler

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Radiopharmaceuticals based in monoclonal anti-body (mAb) via chemical linkers have become a potential tool in nuclear medicine because of their specificity and the large variability and availability of therapeutic radiometals. It is important to identify the conjugation sites and number of attached chelator to mAb to obtain radioimmunoconjugates with required immunoreactivity and radiostability. Girentuximab antibody (G250) is a potential candidate for radioimmunotherapy of clear cell carcinomas (RCCs) because it is reactive with CAIX antigen, a transmembrane glycoprotein overexpressed on the cell surface of most ( > 90%) (RCCs). G250 was conjugated with the bifunctional chelating agent DOTA (1,4,7,10-Tetraazacyclododecane-N,N’,N’’,N’’’-tetraacetic acid) via a benzyl-thiocyano group as a linker (p-SCN-Bn-DOTA). DOTA-G250 conjugates were analyzed by size exclusion chromatography (SE-HPLC) and by electrophoresis (SDS-PAGE). The potential site-specific conjugation was identified by liquid chromatography–mass spectrometry (LC/MS-MS) and the number of linkers per molecule of mAb was calculated using the molecular weight (MW) measured by matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). The average number obtained in the conjugates in non-reduced conditions was between 8-10 molecules of DOTA per molecule of mAb. The average number obtained in the conjugates in reduced conditions was between 1-2 and 3-4 molecules of DOTA per molecule of mAb in the light chain (LC) and heavy chain (HC) respectively. Potential DOTA modification sites of the chelator were identified in lysine residues. The biological activity of the conjugates was evaluated by flow cytometry (FACS) using CAIX negative (SKRC-18) and CAIX positive (SKRC-52). The DOTA-G250 conjugates were labelled with 177Lu with a radiochemical yield > 95% reaching specific activities of 12 MBq/µg. The stability in vitro of different types of radioconstructs was analyzed in human serum albumin (HSA). The radiostability of 177Lu-DOTA-G250 at high specific activity was increased by addition of sodium ascorbate after the labelling. The immunoreactivity was evaluated in vitro and in vivo. Binding to CAIX positive cells (SK-RC-52) at different specific activities was higher for conjugates with less DOTA content. Protein dose was optimized in mice with subcutaneously growing SK-RC-52 tumors using different amounts of 177Lu- DOTA-G250.

Keywords: mass spectrometry, monoclonal antibody, radiopharmaceuticals, radioimmunotheray, renal cancer

Procedia PDF Downloads 307

Authors: Sittampalam Manoharan, Gary Chai, Sanaul Chowdhury, Andrew Golding

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The primary objective of this paper is to present a simplified approach to develop the structural deterioration model using traffic speed deflectometer data for flexible pavements. Maintaining assets to meet functional performance is not economical or sustainable in the long terms, and it would end up needing much more investments for road agencies and extra costs for road users. Performance models have to be included for structural and functional predicting capabilities, in order to assess the needs, and the time frame of those needs. As such structural modelling plays a vital role in the prediction of pavement performance. A structural condition is important for the prediction of remaining life and overall health of a road network and also major influence on the valuation of road pavement. Therefore, the structural deterioration model is a critical input into pavement management system for predicting pavement rehabilitation needs accurately. The Traffic Speed Deflectometer (TSD) is a vehicle-mounted Doppler laser system that is capable of continuously measuring the structural bearing capacity of a pavement whilst moving at traffic speeds. The device’s high accuracy, high speed, and continuous deflection profiles are useful for network-level applications such as predicting road rehabilitations needs and remaining structural service life. The methodology adopted in this model by utilizing time series TSD maximum deflection (D0) data in conjunction with rutting, rutting progression, pavement age, subgrade strength and equivalent standard axle (ESA) data. Then, regression analyses were undertaken to establish a correlation equation of structural deterioration as a function of rutting, pavement age, seal age and equivalent standard axle (ESA). This study developed a simple structural deterioration model which will enable to incorporate available TSD structural data in pavement management system for developing network-level pavement investment strategies. Therefore, the available funding can be used effectively to minimize the whole –of- life cost of the road asset and also improve pavement performance. This study will contribute to narrowing the knowledge gap in structural data usage in network level investment analysis and provide a simple methodology to use structural data effectively in investment decision-making process for road agencies to manage aging road assets.

Keywords: adjusted structural number (SNP), maximum deflection (D0), equant standard axle (ESA), traffic speed deflectometer (TSD)

Procedia PDF Downloads 151

Authors: Yi Li, Rui Lu, Lianjun Wang

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With the rapid development of chemical industry, the consumption of volatile organic compounds (VOCs) has increased extensively. In the process of VOCs production and application, plenty of them have been transferred to environment. As a result, it has led to pollution problems not only in soil and ground water but also to human beings. Thus, it is important to develop a sensitive and cost-effective analytical method for trace VOCs detection in environment. Surface-enhanced Raman Spectroscopy (SERS), as one of the most sensitive optical analytical technique with rapid response, pinpoint accuracy and noninvasive detection, has been widely used for ultratrace analysis. Based on the plasmon resonance on the nanoscale metallic surface, SERS technology can even detect single molecule due to abundant nanogaps (i.e. 'hot spots') on the nanosubstrate. In this work, a self-supported flexible silver nitrate (AgNO3)/ethylene-propylene copolymer (EPM) hybrid nanofibers was fabricated by electrospinning. After an in-situ chemical reduction using ice-cold sodium borohydride as reduction agent, numerous silver nanoparticles were formed on the nanofiber surface. By adjusting the reduction time and AgNO3 content, the morphology and dimension of silver nanoparticles could be controlled. According to the principles of solid-phase extraction, the hydrophobic substance is more likely to partition into the hydrophobic EPM membrane in an aqueous environment while water and other polar components are excluded from the analytes. By the enrichment of EPM fibers, the number of hydrophobic molecules located on the 'hot spots' generated from criss-crossed nanofibers is greatly increased, which further enhances SERS signal intensity. The as-prepared Ag/EPM hybrid nanofibers were first employed to detect common SERS probe molecule (p-aminothiophenol) with the detection limit down to 10-12 M, which demonstrated an excellent SERS performance. To further study the application of the fabricated substrate for monitoring hydrophobic substance in water, several typical VOCs, such as benzene, toluene and p-xylene, were selected as model compounds. The results showed that the characteristic peaks of these target analytes in the mixed aqueous solution could be distinguished even at a concentration of 10-6 M after multi-peaks gaussian fitting process, including C-H bending (850 cm-1), C-C ring stretching (1581 cm-1, 1600 cm-1) of benzene, C-H bending (844 cm-1 ,1151 cm-1), C-C ring stretching (1001 cm-1), CH3 bending vibration (1377 cm-1) of toluene, C-H bending (829 cm-1), C-C stretching (1614 cm-1) of p-xylene. The SERS substrate has remarkable advantages which combine the enrichment capacity from EPM and the Raman enhancement of Ag nanoparticles. Meanwhile, the huge specific surface area resulted from electrospinning is benificial to increase the number of adsoption sites and promotes 'hot spots' formation. In summary, this work provides powerful potential in rapid, on-site and accurate detection of trace VOCs using a portable Raman.

Keywords: electrospinning, ethylene-propylene copolymer, silver nanoparticles, SERS, VOCs

Procedia PDF Downloads 160

Authors: Meiling Yu, Nadia Rouatbi, Khuloud T. Al-Jamal

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Background: Treatment of neurological disorders with modern medical and surgical approaches remains difficult. Gene therapy, allowing the delivery of genetic materials that encodes potential therapeutic molecules, represents an attractive option. The treatment of brain diseases with gene therapy requires the gene-editing tool to be delivered efficiently to the central nervous system. In this study, we explored the efficiency of different delivery routes, namely intravenous (i.v.), intra-cranial (i.c.), and intra-nasal (i.n.), to deliver stable nucleic acid-lipid particles (SNALPs) containing gene-editing tools namely Cas9 mRNA and sgRNA encoding for GFP as a reporter protein. We hypothesise that SNALPs can reach the brain and perform gene-editing to different extents depending on the administration route. Intranasal administration (i.n.) offers an attractive and non-invasive way to access the brain circumventing the blood–brain barrier. Successful delivery of gene-editing tools to the brain offers a great opportunity for therapeutic target validation and nucleic acids therapeutics delivery to improve treatment options for a range of neurodegenerative diseases. In this study, we utilised Rosa26-Cas9 knock-in mice, expressing GFP, to study brain distribution and gene-editing efficiency of SNALPs after i.v.; i.c. and i.n. routes of administration. Methods: Single guide RNA (sgRNA) against GFP has been designed and validated by in vitro nuclease assay. SNALPs were formulated and characterised using dynamic light scattering. The encapsulation efficiency of nucleic acids (NA) was measured by RiboGreen™ assay. SNALPs were incubated in serum to assess their ability to protect NA from degradation. Rosa26-Cas9 knock-in mice were i.v., i.n., or i.c. administered with SNALPs to test in vivo gene-editing (GFP knockout) efficiency. SNALPs were given as three doses of 0.64 mg/kg sgGFP following i.v. and i.n. or a single dose of 0.25 mg/kg sgGFP following i.c.. knockout efficiency was assessed after seven days using Sanger Sequencing and Inference of CRISPR Edits (ICE) analysis. In vivo, the biodistribution of DiR labelled SNALPs (SNALPs-DiR) was assessed at 24h post-administration using IVIS Lumina Series III. Results: Serum-stable SNALPs produced were 130-140 nm in diameter with ~90% nucleic acid loading efficiency. SNALPs could reach and stay in the brain for up to 24h following i.v.; i.n. and i.c. administration. Decreasing GFP expression (around 50% after i.v. and i.c. and 20% following i.n.) was confirmed by optical imaging. Despite the small number of mice used, ICE analysis confirmed GFP knockout in mice brains. Additional studies are currently taking place to increase mice numbers. Conclusion: Results confirmed efficient gene knockout achieved by SNALPs in Rosa26-Cas9 knock-in mice expressing GFP following different routes of administrations in the following order i.v.= i.c.> i.n. Each of the administration routes has its pros and cons. The next stages of the project involve assessing gene-editing efficiency in wild-type mice and replacing GFP as a model target with therapeutic target genes implicated in Motor Neuron Disease pathology.

Keywords: CRISPR, nanoparticles, brain diseases, administration routes

Procedia PDF Downloads 102

Authors: Fatna Benmessaoud, Mohammed Cheikh, Vencent Velay, Vanessa Vedal, Farhad Rezai-Aria, Christine Boher

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The global mechanical behavior of materials is strongly linked to their microstructure, especially their crystallographic texture and their grains morphology. These material aspects determine the mechanical fields character (heterogeneous or homogeneous), thus, they give to the global behavior a degree of anisotropy according the initial microstructure. For these reasons, the prediction of global behavior of materials in relationship with the microstructure must be performed with a multi-scale approach. Therefore, multi-scale modeling in the context of crystal plasticity is widely used. In this present contribution, a phenomenological elasto-viscoplastic model developed in the crystal plasticity context and finite element method are used to investigate the effects of crystallographic texture and grains sizes on global behavior of a polycrystalline equiaxed Ti-6Al-4V alloy. The constitutive equations of this model are written on local scale for each slip system within each grain while the strain and stress mechanical fields are investigated at the global scale via finite element scale transition. The beta phase of Ti-6Al-4V alloy modeled is negligible; its percent is less than 10%. Three families of slip systems of alpha phase are considered: basal and prismatic families with a burgers vector and pyramidal family with a burgers vector. The twinning mechanism of plastic strain is not observed in Ti-6Al-4V, therefore, it is not considered in the present modeling. Nine representative elementary volumes (REV) are generated with Voronoi tessellations. For each individual equiaxed grain, the own crystallographic orientation vis-à-vis the loading is taken into account. The meshing strategy is optimized in a way to eliminate the meshing effects and at the same time to allow calculating the individual grain size. The stress and strain fields are determined in each Gauss point of the mesh element. A post-treatment is used to calculate the local behavior (in each grain) and then by appropriate homogenization, the macroscopic behavior is calculated. The developed model is validated by comparing the numerical simulation results with an experimental data reported in the literature. It is observed that the present model is able to predict the global mechanical behavior of Ti-6Al-4V alloy and investigate the microstructural parameters' effects. According to the simulations performed on the generated volumes (REV), the macroscopic mechanical behavior of Ti-6Al-4V is strongly linked to the active slip systems family (prismatic, basal or pyramidal). The crystallographic texture determines which family of slip systems can be activated; therefore it gives to the plastic strain a heterogeneous character thus an anisotropic macroscopic mechanical behavior. The average grains size influences also the Ti-6Al-4V mechanical proprieties, especially the yield stress; by decreasing of the average grains size, the yield strength increases according to Hall-Petch relationship. The grains sizes' distribution gives to the strain fields considerable heterogeneity. By increasing grain sizes, the scattering in the localization of plastic strain is observed, thus, in certain areas the stress concentrations are stronger than other regions.

Keywords: microstructural parameters, multi-scale modeling, crystal plasticity, Ti-6Al-4V alloy

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Authors: M. Maleczek, Leszek Bogdan, Kazimierz Drabczyk, Agnieszka Iwan

Abstract:

Crystalline silicon (Si) solar cells are the main product in the market among the various photovoltaic technologies concerning such advantages as: material richness, high carrier mobilities, broad spectral absorption range and established technology. However, photovoltaic technology on the stiff substrates are heavier, more fragile and less cost-effective than devices on the flexible substrates to be applied in special applications. The main goal of our work was to incorporate silicon solar cells into various fabric, without any change of the electrical and mechanical parameters of devices. This work is realized for the GEKON project (No. GEKON2/O4/268473/23/2016) sponsored by The National Centre for Research and Development and The National Fund for Environmental Protection and Water Management. In our work, the polyamide or polyester fabrics were used as a flexible substrate in the created devices. Applied fabrics differ in tensile and tear strength. All investigated polyamide fabrics are resistant to weathering and UV, while polyester ones is resistant to ozone, water and ageing. The examined fabrics are tight at 100 cm water per 2 hours. In our work, commercial silicon solar cells with the size 156 × 156 mm were cut into nine parts (called single solar cells) by diamond saw and laser. Gap and edge after cutting of solar cells were checked by transmission electron microscope (TEM) to study morphology and quality of the prepared single solar cells. Modules with the size of 160 × 70 cm (containing about 80 single solar cells) were created and investigated by electrical and mechanical methods. Weight of constructed module is about 1.9 kg. Three types of solar cell architectures such as: -fabric/EVA/Si solar cell/EVA/film for lamination, -backsheet PET/EVA/Si solar cell/EVA/film for lamination, -fabric/EVA/Si solar cell/EVA/tempered glass, were investigated taking into consideration type of fabric and lamination process together with the size of solar cells. In investigated devices EVA, it is ethylene-vinyl acetate, while PET - polyethylene terephthalate. Depend on the lamination process and compatibility of textile with solar cell an efficiency of investigated flexible silicon solar cells was in the range of 9.44-16.64 %. Multi folding and unfolding of flexible module has no impact on its efficiency as was detected by Instron equipment. Power (P) of constructed solar module is 30 W, while voltage about 36 V. Finally, solar panel contains five modules with the polyamide fabric and tempered glass will be produced commercially for different applications (dual use).

Keywords: flexible devices, mechanical properties, silicon solar cells, textiles

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Authors: Huan Peng, Chenye Zeng, Zhao Wang

Abstract:

Alzheimer’s disease (AD) is an age-related neurodegenerative disease that is a leading cause of dementia syndromes and has become a huge burden on society and families. The main pathological features of AD involve excessive deposition of β-amyloid (Aβ) and Tau proteins in the brain, resulting in loss of neurons, expansion of neuroinflammation, and cognitive dysfunction in patients. Researchers have found effective drugs to clear the brain of error-accumulating proteins or to slow the loss of neurons, but their direct administration has key bottlenecks such as single-drug limitation, rapid blood clearance rate, impenetrable blood-brain barrier (BBB), and poor ability to target tissues and cells. Therefore, we are committed to seeking a suitable and efficient delivery system. Inspired by the possibility that exosomes may be involved in the secretion and transport mechanism of many signaling molecules or proteins in the brain, exosomes have attracted extensive attention as natural nanoscale drug carriers. We selected exosomes derived from bone marrow mesenchymal stem cells (MSC-EXO) with low immunogenicity and exosomes derived from hippocampal neurons (HT22-EXO) that may have excellent homing ability to overcome the deficiencies of oral or injectable pathways and bypass the BBB through nasal administration and evaluated their delivery ability and effect on AD. First, MSC-EXO and HT22 cells were isolated and cultured, and MSCs were identified by microimaging and flow cytometry. Then MSC-EXO and HT22-EXO were obtained by gradient centrifugation and qEV SEC separation column, and a series of physicochemical characterization were performed by transmission electron microscope, western blot, nanoparticle tracking analysis and dynamic light scattering. Next, exosomes labeled with lipophilic fluorescent dye were administered to WT mice and APP/PS1 mice to obtain fluorescence images of various organs at different times. Finally, APP/PS1 mice were administered intranasally with two exosomes 20 times over 40 days and 20 μL each time. Behavioral analysis and pathological section analysis of the hippocampus were performed after the experiment. The results showed that MSC-EXO and HT22-EXO were successfully isolated and characterized, and they had good biocompatibility. MSC-EXO showed excellent brain enrichment in APP/PS1 mice after intranasal administration, could improve the neuronal damage and reduce inflammation levels in the hippocampus of APP/PS1 mice, and the improvement effect was significantly better than HT22-EXO. However, intranasal administration of the two exosomes did not cause depression and anxious-like phenotypes in APP/PS1 mice, nor significantly improved the short-term or spatial learning and memory ability of APP/PS1 mice, and had no significant effect on the content of Aβ plaques in the hippocampus, which also meant that MSC-EXO could use their own advantages in combination with other drugs to clear Aβ plaques. The possibility of realizing highly effective non-invasive synergistic treatment for AD provides new strategies and ideas for clinical research.

Keywords: Alzheimer’s disease, exosomes derived from mesenchymal stem cell, intranasal administration, therapy-carrier dual roles

Procedia PDF Downloads 62

Authors: Benson Tette, Thomas Mensah

Abstract:

As a developing country, Ghana has a high potential to harness the economic value of every industry. Two of the industries that produce below capacity are handicrafts (for instance, carpentry) and information technology (i.e., computer science). To boost production and maintain competitiveness, the carpentry sector in Ghana needs more effective manufacturing procedures that are also more affordable. This issue can be resolved using computer-aided manufacturing (CAM) technology, which automates the fabrication process and decreases the amount of time and labor needed to make wood goods. Yet, the integration of CAM in carpentry-related production is rarely explored. To streamline the manufacturing process, this research investigates the equipment and technology that are currently used in the Ghanaian carpentry sector for automated fabrication. The research looks at the various CAM technologies, such as Computer Numerical Control routers, laser cutters, and plasma cutters, that are accessible to Ghanaian carpenters yet unexplored. We also investigate their potential to enhance the production process. To achieve the objective, 150 carpenters, 15 software engineers, and 10 policymakers were interviewed using structured questionnaires. The responses provided by the 175 respondents were processed to eliminate outliers and omissions were corrected using multiple imputations techniques. The processed responses were analyzed through thematic analysis. The findings showed that adaptation and integration of CAD software with CAM technologies would speed up the design-to-manufacturing process for carpenters. It must be noted that achieving such results entails first; examining the capabilities of current CAD software, then determining what new functions and resources are required to improve the software's suitability for carpentry tasks. Responses from both carpenters and computer scientists showed that it is highly practical and achievable to streamline the design-to-manufacturing process through processes such as modifying and combining CAD software with CAM technology. Making the carpentry-software integration program more useful for carpentry projects would necessitate investigating the capabilities of the current CAD software and identifying additional features in the Ghanaian ecosystem and tools that are required. In conclusion, the Ghanaian carpentry sector has a chance to increase productivity and competitiveness through the integration of CAM technology with CAD software. Carpentry companies may lower labor costs and boost production capacity by automating the fabrication process, giving them a competitive advantage. This study offers implementation-ready and representative recommendations for successful implementation as well as important insights into the equipment and technologies available for automated fabrication in the Ghanaian carpentry sector.

Keywords: carpentry, computer-aided manufacturing (CAM), Ghana, information technology(IT)

Procedia PDF Downloads 98

Authors: Bruno Vicente, Sandra Rafael, Vera Rodrigues, Sandra Sorte, Sara Silva, Ana Isabel Miranda, Carlos Borrego

Abstract:

Wind comfort for pedestrians is an important condition in urban areas. In Portugal, a country with 900 km of coastline, the wind direction are predominantly from Nor-Northwest with an average speed of 2.3 m·s -1 (at 2 m height). As a result, a set of city authorities have been requesting studies of pedestrian wind comfort for new urban areas/buildings, as well as to mitigate wind discomfort issues related to existing structures. This work covers the efficiency evaluation of a set of measures to reduce the wind speed in an outdoor auditorium (open space) located in a coastal Portuguese urban area. These measures include the construction of barriers, placed at upstream and downstream of the auditorium, and the planting of trees, placed upstream of the auditorium. The auditorium is constructed in the form of a porch, aligned with North direction, driving the wind flow within the auditorium, promoting channelling effects and increasing its speed, causing discomfort in the users of this structure. To perform the wind comfort assessment, two approaches were used: i) a set of experiments using the wind tunnel (physical approach), with a representative mock-up of the study area; ii) application of the CFD (Computational Fluid Dynamics) model VADIS (numerical approach). Both approaches were used to simulate the baseline scenario and the scenarios considering a set of measures. The physical approach was conducted through a quantitative method, using hot-wire anemometer, and through a qualitative analysis (visualizations), using the laser technology and a fog machine. Both numerical and physical approaches were performed for three different velocities (2, 4 and 6 m·s-1 ) and two different directions (NorNorthwest and South), corresponding to the prevailing wind speed and direction of the study area. The numerical results show an effective reduction (with a maximum value of 80%) of the wind speed inside the auditorium, through the application of the proposed measures. A wind speed reduction in a range of 20% to 40% was obtained around the audience area, for a wind direction from Nor-Northwest. For southern winds, in the audience zone, the wind speed was reduced from 60% to 80%. Despite of that, for southern winds, the design of the barriers generated additional hot spots (high wind speed), namely, in the entrance to the auditorium. Thus, a changing in the location of the entrance would minimize these effects. The results obtained in the wind tunnel compared well with the numerical data, also revealing the high efficiency of the purposed measures (for both wind directions).

Keywords: urban microclimate, pedestrian comfort, numerical modelling, wind tunnel experiments

Procedia PDF Downloads 230