Search results for: prototype fuel cell electric vehicles
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 7640

Search results for: prototype fuel cell electric vehicles

5570 Biodiesel Production from Yellow Oleander Seed Oil

Authors: S. Rashmi, Devashish Das, N. Spoorthi, H. V. Manasa

Abstract:

Energy is essential and plays an important role for overall development of a nation. The global economy literally runs on energy. The use of fossil fuels as energy is now widely accepted as unsustainable due to depleting resources and also due to the accumulation of greenhouse gases in the environment, renewable and carbon neutral biodiesel are necessary for environment and economic sustainability. Unfortunately biodiesel produced from oil crop, waste cooking oil and animal fats are not able to replace fossil fuel. Fossil fuels remain the dominant source of primary energy, accounting for 84% of the overall increase in demand. Today biodiesel has come to mean a very specific chemical modification of natural oils. Objectives: To produce biodiesel from yellow oleander seed oil, to test the yield of biodiesel using different types of catalyst (KOH & NaOH). Methodology: Oil is extracted from dried yellow oleander seeds using Soxhlet extractor and oil expeller (bulk). The FFA content of the oil is checked and depending on the FFA value either two steps or single step process is followed to produce biodiesel. Two step processes includes esterfication and transesterification, single step includes only transesterification. The properties of biodiesel are checked. Engine test is done for biodiesel produced. Result: It is concluded that biodiesel quality parameters such as yield(85% & 90%), flash point(1710C & 1760C),fire point(1950C & 1980C), viscosity(4.9991 and 5.21 mm2/s) for the biodiesel from seed oil of Thevetiaperuviana produced by using KOH & NaOH respectively. Thus the seed oil of Thevetiaperuviana is a viable feedstock for good quality fuel.The outcomes of our project are a substitute for conventional fuel, to reduce petro diesel requirement,improved performance in terms of emissions. Future prospects: Optimization of biodiesel production using response surface method.

Keywords: yellow oleander seeds, biodiesel, quality parameters, renewable sources

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5569 Determination of Aflatoxins in Edible-Medicinal Plant Samples by HPLC with Fluorescence Detector and KOBRA-Cell

Authors: Isil Gazioglu, Abdulselam Ertas

Abstract:

Aflatoxins (AFs) are secondary toxic metabolites of Aspergillus flavus and A. parasiticus. AFs can be absorbed through the skin. Potent carcinogens like AFs should be completely absent from cosmetics, this can be achieved by careful quality control of the raw plant materials. Regulatory limits for aflatoxins have been established in many countries, and reliable testing methodology is needed to implement and enforce the regulatory limits. In this study, ten medicinal plant samples (Bundelia tournefortti, Capsella bursa-pastoris, Carduus tenuiflorus, Cardaria draba, Malva neglecta, Malvella sharardiana, Melissa officinalis, Sideritis libanotica, Stakys thirkei, Thymus nummularius) were investigated for aflatoxin (AF) contaminations by employing an HPLC assay for the determination of AFB1, B2, G1 and G2. The samples were extracted with 70% (v/v) methanol in water before further cleaned up with an immunoaffinity column and followed by the detection of AFs by using an electrochemically post-column derivatization with Kobra-Cell and fluorescence detector. The extraction procedure was optimized in order to obtain the best recovery. The method was successfully carried out with all medicinal plant samples. The results revealed that five (50%) of samples were contaminated with AFs. The association between particular samples and the AF contaminated could not be determined due to the low frequency of positive samples.

Keywords: aflatoxin B1, HPLC-FLD, KOBRA-Cell, mycotoxin

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5568 Techno Commercial Aspects of Using LPG as an Alternative Energy Solution for Transport and Industrial Sector in Bangladesh: Case Studies in Industrial Sector

Authors: Mahadehe Hassan

Abstract:

Transport system and industries which are the main basis of industrial and socio-economic development of any country. It is mainly dependent on fossil fuels. Bangladesh has fossil fuel reserves of 9.51 TCF as of July 2023, and if no new gas fields are discovered in the next 7-9 years and if the existing gas consumption rate continues, the fossil fuel reserves will be exhausted. The demand for petroleum products in Bangladesh is increasing steadily, with 63% imported by BPC and 37% imported by private companies. 61.61% of BPC imported products are used in the transport sector and 5.49% in the industrial sector, which is expensive and harmful to the environment. Liquefied Petroleum Gas (LPG) should be considered as an alternative energy for Bangladesh based on Sustainable Development Goals (SDGs) criteria for sustainable, clean and affordable energy. This will not only lead to the much desired mitigation of energy famine in the country but also contribute favorably to the macroeconomic indicators. Considering the environmental and economic issues, the government has referred to CNG (compressed natural gas) as the fuel carrier since 2000, but currently due to the decline mode of gas reserves, the government of Bangladesh is thinking of new energy sources for transport and industrial sectors which will be sustainable, environmentally friendly and economically viable. Liquefied Petroleum Gas (LPG) is the best choice for fueling transport and industrial sectors in Bangladesh. At present, a total of 1.54 million metric tons of liquefied petroleum gas (LPG) is marketed in Bangladesh by the public and private sectors. 83% of it is used by households, 12% by industry and commerce and 5% by transportation. Industrial and transport sector consumption is negligible compared to household consumption. So the purpose of the research is to find out the challenges of LPG market development in transport and industrial sectors in Bangladesh and make recommendations to reduce the challenges. Secure supply chain, inadequate infrastructure, insufficient investment, lack of government monitoring and consumer awareness in the transport sector and industrial sector are major challenges for LPG market development in Bangladesh. Bangladesh government as well as private owners should come forward in the development of liquefied petroleum gas (LPG) industry to reduce the challenges of secure energy sector for sustainable development. Furthermore, ensuring adequate Liquefied Petroleum Gas (LPG) supply in Bangladesh requires government regulations, infrastructure improvements in port areas, awareness raising and most importantly proper pricing of Liquefied Petroleum Gas (LPG) to address the energy crisis in Bangladesh.

Keywords: transportand industries fuel, LPG consumption, challenges, economical sustainability

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5567 The Optimum Biodiesel Blend in Low Sulfur Diesel and Its Physico-Chemical Properties and Economic Aspect

Authors: Ketsada Sutthiumporn, Sittichot Thongkaw, Malee Santikunaporn

Abstract:

In Thailand, biodiesel has been utilized as an attractive substitute of petroleum diesel and the government imposes a mandatory biodiesel blending requirement in transport sector to improve energy security, support agricultural sector and reduce emissions. Though biodiesel blend has many advantages over diesel fuel such as improved lubricity, low sulfur content and higher flash point, there are still some technical problems such as oxidative stability, poor cold- flow properties and impurity. Such problems were related to the fatty acid composition in feedstock. Moreover, Thailand has announced the use of low sulfur diesel as a base diesel and will be continually upgrading to EURO 5 in 2023. With ultra low sulfur content, it may affect the diesel fuel properties especially lubricity as well. Therefore, in this study, the physical and chemical properties of palm oil-based biodiesel in low sulfur diesel blends from different producers will be investigated by standard methods per ASTM and EN. Also, its economic benefits based on diesel price structure in Thailand will be highlighted. The appropriate biodiesel blend ratio can affect the physico-chemical properties and reasonable price in the country. Properties of biodiesel, including specific gravity, kinematic viscosity, FAME composition, flash point, sulfur, water, oxidation stability and lubricity were measured by standard methods of ASTM and EN. The results show that the FAME composition of biodiesel has the fatty acid of C12:0 to C20:1, mostly in C16:0, C18:0, C18:1, and C18:2, which were main characteristic compositions of palm biodiesel. The physical and chemical properties of biodiesel blended diesel was found to be increases with an increasing amount of biodiesel such as specific gravity, flash point and kinematic viscosity while sulfur value was decreased. Moreover, in this study, the various properties of each biodiesel blends were plotted to determine the appropriate proportional range of biodiesel-blended diesel with an optimum fuel price.It can be seen that the amount of B100 can be filled from 1% up to 7% in which the quality was in accordance with Notification of the department of Energy business.The understanding of relation between physico-chemical properties of palm oil-based biodiesel and pricing is beneficial to guide the better development of desired feedstock in Thailand and to implement biodiesel blends with comparative price and diesel engine performance.

Keywords: fatty acid methyl ester, biodiesel, fuel price structure, palm oil in Thailand

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5566 Convolutional Neural Network and LSTM Applied to Abnormal Behaviour Detection from Highway Footage

Authors: Rafael Marinho de Andrade, Elcio Hideti Shiguemori, Rafael Duarte Coelho dos Santos

Abstract:

Relying on computer vision, many clever things are possible in order to make the world safer and optimized on resource management, especially considering time and attention as manageable resources, once the modern world is very abundant in cameras from inside our pockets to above our heads while crossing the streets. Thus, automated solutions based on computer vision techniques to detect, react, or even prevent relevant events such as robbery, car crashes and traffic jams can be accomplished and implemented for the sake of both logistical and surveillance improvements. In this paper, we present an approach for vehicles’ abnormal behaviors detection from highway footages, in which the vectorial data of the vehicles’ displacement are extracted directly from surveillance cameras footage through object detection and tracking with a deep convolutional neural network and inserted into a long-short term memory neural network for behavior classification. The results show that the classifications of behaviors are consistent and the same principles may be applied to other trackable objects and scenarios as well.

Keywords: artificial intelligence, behavior detection, computer vision, convolutional neural networks, LSTM, highway footage

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5565 Developing a Model of Teaching Writing Based On Reading Approach through Reflection Strategy for EFL Students of STKIP YPUP

Authors: Eny Syatriana, Ardiansyah

Abstract:

The purpose of recent study was to develop a learning model on writing, based on the reading texts which will be read by the students using reflection strategy. The strategy would allow the students to read the text and then they would write back the main idea and to develop the text by using their own sentences. So, the writing practice was begun by reading an interesting text, then the students would develop the text which has been read into their writing. The problem questions are (1) what kind of learning model that can develop the students writing ability? (2) what is the achievement of the students of STKIP YPUP through reflection strategy? (3) is the using of the strategy effective to develop students competence In writing? (4) in what level are the students interest toward the using of a strategy In writing subject? This development research consisted of some steps, they are (1) need analysis (2) model design (3) implementation (4) model evaluation. The need analysis was applied through discussion among the writing lecturers to create a learning model for writing subject. To see the effectiveness of the model, an experiment would be delivered for one class. The instrument and learning material would be validated by the experts. In every steps of material development, there was a learning process, where would be validated by an expert. The research used development design. These Principles and procedures or research design and development .This study, researcher would do need analysis, creating prototype, content validation, and limited empiric experiment to the sample. In each steps, there should be an assessment and revision to the drafts before continue to the next steps. The second year, the prototype would be tested empirically to four classes in STKIP YPUP for English department. Implementing the test greatly was done through the action research and followed by evaluation and validation from the experts.

Keywords: learning model, reflection, strategy, reading, writing, development

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5564 Coated Chromium Thin Film on Zirconium for Corrosion Resistance of Nuclear Fuel Rods by Plasma Focus Device

Authors: Amir Raeisdana, Davood Sohrabi, Mojtaba Nohekhan, Ameneh Kargarian, Maryam Ghapanvari, Alireza Aslezaeem

Abstract:

Improvement of zirconium properties by chromium coating and nitrogen implantation is ideal to protect the nuclear fuel rods against corrosion and secondary hydrogenation. Metallic chromium (Cr) has attracted attention as a potential coating material on zirconium alloys, to limit external cladding corrosion. In this research, high energy plasma focus device was used to coat the chromium and implant the nitrogen ions in the zirconium substrate. This device emits high-energy nitrogen ions of 10 keV-1 MeV and with a flux of 10^16 ions/cm^2 in each shot toward the target so it is attractive for implantation on the substrate materials at the room temperature. Six zirconium samples in 2cm×2cm dimensions with 1mm thickness were located at a distance of 20cm from the place where the pinch is formed. The experiments are carried out in 0.5 mbar of the nitrogen gas pressure and 15 kV of the charging voltage. Pure Cr disc was installed on the anode head for sputtering of the chromium and deposition on zirconium substrate. When the pinch plasma column decays due to various instabilities, intense and high-energy N2 ions are accelerated towards the zirconium substrate also sputtered Cr is deposited on the zirconium substrate. XRD and XRF analysis were used to study the structural properties of the samples. XRF analysis indicates 77.1% of Zr and 11.1% of Cr in the surface of the sample. XRD spectra shows the formation of ZrN, CrN and CrZr composites after nitrogen implantation and chromium coating. XRD spectra shows the chromium peak height equal to 152.80 a.u. for the major sample (θ=0֯) and 92.99 a.u. for the minor sample (θ=6֯), so implantation and coating along the main axis of the device is significantly more than other directions.

Keywords: ZrN and CrN and CrZr composites, angular distribution for Cr deposition rate, zirconium corrosion resistance, nuclear fuel rods, plasma focus device

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5563 Hysteresis Effect in Organometallic Perovskite Solar Cells with Mesoscopic NiO as a Hole Transport Layer

Authors: D. C. Asebiah, D. Saranin, S. Karazhanov, A. R. Tameev, M. Kah

Abstract:

In this paper, the mesoscopic NiO was used as a hole transport layer in the inverted planar organometallic hybrid perovskite solar cell to study the effect of hysteresis. The devices we fabricated have the structures Fluorine Tin Oxide (FTO)/mesoscopic NiO/perovskite/[6,6]-phenyl C₆₁-butyric acid methyl ester (PC₆₁BM) photovoltaic device. The perovskite solar cell was done by toluene air (TLA) method and horn sonication for the dispersion of the NiO nanoparticles in deionized water. The power conversion efficiency was 12.07% under 1.5 AM illumination. We report hysteresis in the in current-voltage dependence of the solar cells with mesoscopic NiO as a hole transport layer.

Keywords: perovskite, mesoscopic, hysteresis, toluene air

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5562 The Impact of Ultrasonicator on the Vertical and Horizontal Mixing Profile of Petrol-Bioethanol

Authors: D. Nkazi, S. E. Iyuke, J. Mulopo

Abstract:

Increasing global energy demand as well as air quality concerns have in recent years led to the search for alternative clean fuels to replace fossil fuels. One such alternative is the blending of petrol with ethanol, which has numerous advantages such ethanol’s ability to act as oxygenate thus reducing the carbon monoxide emissions from the exhaust of internal combustion engines of vehicles. However, the hygroscopic nature of ethanol is a major concern in obtaining a perfectly homogenized petrol-ethanol fuel. This problem has led to the study of ways of homogenizing the petrol-ethanol mixtures. During the blending process, volumes fraction of ethanol and petrol were studied with respect to the depth within the storage container to confirm homogenization of the blend and time of storage. The results reveal that the density of the mixture was constant. The binodal curve of the ternary diagram shows an increase of homogeneous region, indicating an improved of interaction between water and petrol. The concentration distribution in the reactor showed proof of cavitation formation since in both directions, the variation of concentration with both time and distance was found to be oscillatory. On comparing the profiles in both directions, the concentration gradient, diffusion flux, and energy and diffusion rates were found to be higher in the vertical direction compared to the horizontal direction. It was therefore concluded that ultrasonication creates cavitation in the mixture which enhances mass transfer and mixing of ethanol and petrol. The horizontal direction was found to be the diffusion rate limiting step which proposed that the blender should have a larger height to diameter ratio. It is, however, recommended that further studies be done on the rate-limiting step so as to have actual dimensions of the reactor.

Keywords: ultrasonication, petrol, ethanol, concentration

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5561 A Dissipative Particle Dynamics Study of a Capsule in Microfluidic Intracellular Delivery System

Authors: Nishanthi N. S., Srikanth Vedantam

Abstract:

Intracellular delivery of materials has always proved to be a challenge in research and therapeutic applications. Usually, vector-based methods, such as liposomes and polymeric materials, and physical methods, such as electroporation and sonoporation have been used for introducing nucleic acids or proteins. Reliance on exogenous materials, toxicity, off-target effects was the short-comings of these methods. Microinjection was an alternative process which addressed the above drawbacks. However, its low throughput had hindered its adoption widely. Mechanical deformation of cells by squeezing them through constriction channel can cause the temporary development of pores that would facilitate non-targeted diffusion of materials. Advantages of this method include high efficiency in intracellular delivery, a wide choice of materials, improved viability and high throughput. This cell squeezing process can be studied deeper by employing simple models and efficient computational procedures. In our current work, we present a finite sized dissipative particle dynamics (FDPD) model to simulate the dynamics of the cell flowing through a constricted channel. The cell is modeled as a capsule with FDPD particles connected through a spring network to represent the membrane. The total energy of the capsule is associated with linear and radial springs in addition to constraint of the fixed area. By performing detailed simulations, we studied the strain on the membrane of the capsule for channels with varying constriction heights. The strain on the capsule membrane was found to be similar though the constriction heights vary. When strain on the membrane was correlated to the development of pores, we found higher porosity in capsule flowing in wider channel. This is due to localization of strain to a smaller region in the narrow constriction channel. But the residence time of the capsule increased as the channel constriction narrowed indicating that strain for an increased time will cause less cell viability.

Keywords: capsule, cell squeezing, dissipative particle dynamics, intracellular delivery, microfluidics, numerical simulations

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5560 Numerical Investigation of Electrohydrodynamics: Enhanced Heat Transfer in a Solid Sample

Authors: Suwimon Saneewong Na Ayuttaya

Abstract:

This paper presents a numerical investigation of electrically driven flow for enhancing convective heat transfer in a channel flow. This study focuses on the electrode arrangements, number of electrode and electrical voltage on Electrohydrodynamics (EHD) and effect of airflow driven on solid sample surface. The inlet airflow and inlet temperature are 0.35 m/s and 60 oC, respectively. High electrical voltage is tested in the range of 0-30 kV and number of electrode is tested in the range of 1-5. The numerical results show that electric field intensity is depended on electrical voltage and number of electrode. Increasing number of electrodes is increased shear flow, so swirling flow is increased. The swirling flows from aligned and staggered arrangements are affecting within the solid sample. When electrical voltage is increased, temperature distribution and convective heat transfer on the solid sample are significantly increased due to the electric force much stronger.

Keywords: electrohydrodynamics (EHD), swirling flow, convective heat transfer, solid sample

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5559 Protective Coating Layers via Phosphazene Compounds for Stabilizing Silicon Anode Materials

Authors: Adjmal Ghaur, Christoph Peschel, Iris Dienwiebel, Lukas Haneke, Leilei Du , Laurin Profanter, Tobias Placke, Martin Winter

Abstract:

In recent years, lithium-ion batteries (LIBs)are widely used in electric vehicles (EVs) and mobile energy storage devices (ESDs), which has led to higher requirements for energy density. To fulfill these requirements, tremendous attention has been paid to design advanced LIBs with various siliconactive materials as alternative negative electrodes to replace graphite (372 mAh g⁻¹)due to their high theoretical gravimetric capacity (4200mAh g⁻¹). However, silicon as potential anode material suffers from huge volume changes during charging and discharging and has poor electronicconductivity which negatively impacts the long-term performance and preventshigh silicon contents from practical application. Additionally, an unstable crystalline silicon structure tends to pulverization during the (de)lithiation process. To compensate for the volume changes, alleviate pulverization, and maintain high electronicconductivity, silicon-doped graphite composites with protecting coating layers are a promising approach. In this context, phosphazene compounds are investigated concerning their silicon protecting properties in silicon-doped graphite composites. In detail, electrochemical performance measurements in pouch full-cells(NCM523||SiOx/C), supressing gas formation properties, and post-mortem analyzes were carried out to characterize phosphazene compounds as additive materials. The introduction of the dual-additive approach in state-of-the-art electrolytes leads to synergistic effects between FEC and phosphazene compounds which accelerate the durability of silicon particles and results in enhanced electrochemical performance.

Keywords: silicon, phosphazene, solid electrolyte interphase, electrolyte, gasmeasurements

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5558 Development of Noninvasive Method to Analyze Dynamic Changes of Matrix Stiffness and Elasticity Characteristics

Authors: Elena Petersen, Inna Kornienko, Svetlana Guryeva, Sergey Dobdin, Anatoly Skripal, Andrey Usanov, Dmitry Usanov

Abstract:

One of the most important unsolved problems in modern medicine is the increase of chronic diseases that lead to organ dysfunction or even complete loss of function. Current methods of treatment do not result in decreased mortality and disability statistics. Currently, the best treatment for many patients is still transplantation of organs and/or tissues. Therefore, finding a way of correct artificial matrix biofabrication in case of limited number of natural organs for transplantation is a critical task. One important problem that needs to be solved is development of a nondestructive and noninvasive method to analyze dynamic changes of mechanical characteristics of a matrix with minimal side effects on the growing cells. This research was focused on investigating the properties of matrix as a marker of graft condition. In this study, the collagen gel with human primary dermal fibroblasts in suspension (60, 120, 240*103 cells/mL) and collagen gel with cell spheroids were used as model objects. The stiffness and elasticity characteristics were evaluated by a semiconductor laser autodyne. The time and cell concentration dependency of the stiffness and elasticity were investigated. It was shown that these properties changed in a non-linear manner with respect to cell concentration. The maximum matrix stiffness was observed in the collagen gel with the cell concentration of 120*103 cells/mL. This study proved the opportunity to use the mechanical properties of matrix as a marker of graft condition, which can be measured by noninvasive semiconductor laser autodyne technique.

Keywords: graft, matrix, noninvasive method, regenerative medicine, semiconductor laser autodyne

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5557 Performance of the Photovoltaic Module under Different Shading Patterns

Authors: E. T. El Shenawy, O. N. A. Esmail, Adel A. Elbaset, Hesham F. A. Hamed

Abstract:

Generation of the electrical energy based on photovoltaic (PV) technology has been increased over the world due to either the continuous reduction in the traditional energy sources in addition to the pollution problems related to their usage, or the clean nature and safe usage of the PV technology. Also, PV systems can generate clean electricity in the site of use without any transmission, which can be considered cost effective than other generation systems. The performance of the PV system is highly affected by the amount of solar radiation incident on it. Completely or partially shaded PV systems can affect its output. The PV system can be shaded by trees, buildings, dust, incorrect system configuration, or other obstacles. The present paper studies the effect of the partial shading on the performance of a thin film PV module under climatic conditions of Cairo, Egypt. This effect was measured and evaluated according to practical measurement of the characteristic curves such as current-voltage and power-voltage for two identical PV modules (with and without shading) placed at the same time on one mechanical structure for comparison. The measurements have been carried out for the following shading patterns; half cell (bottom, middle, and top of the PV module); complete cell; and two adjacent cells. The results showed that partially shading the PV module changes the shapes of the I-V and P-V curves and produces more than one maximum power point, that can disturb the traditional maximum power point trackers. Also, the output power from the module decreased according to the incomplete solar radiation reaching the PV module due to shadow patterns. The power loss due shading was 7%, 22%, and 41% for shading of half-cell, one cell, and two adjacent cells of the PV module, respectively.

Keywords: I-V measurements, PV module characteristics, PV module power loss, PV module shading

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5556 Comparative Study of Scheduling Algorithms for LTE Networks

Authors: Samia Dardouri, Ridha Bouallegue

Abstract:

Scheduling is the process of dynamically allocating physical resources to User Equipment (UE) based on scheduling algorithms implemented at the LTE base station. Various algorithms have been proposed by network researchers as the implementation of scheduling algorithm which represents an open issue in Long Term Evolution (LTE) standard. This paper makes an attempt to study and compare the performance of PF, MLWDF and EXP/PF scheduling algorithms. The evaluation is considered for a single cell with interference scenario for different flows such as Best effort, Video and VoIP in a pedestrian and vehicular environment using the LTE-Sim network simulator. The comparative study is conducted in terms of system throughput, fairness index, delay, packet loss ratio (PLR) and total cell spectral efficiency.

Keywords: LTE, multimedia flows, scheduling algorithms, mobile computing

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5555 Short-Term Association of In-vehicle Ultrafine Particles and Black Carbon Concentrations with Respiratory Health in Parisian Taxi Drivers

Authors: Melissa Hachem, Maxime Loizeau, Nadine Saleh, Isabelle Momas, Lynda Bensefa-Colas

Abstract:

Professional drivers are exposed inside their vehicles to high levels of air pollutants due to the considerable time they spend close to motor vehicle emissions. Little is known about ultrafine particles (UFP) or black carbon (BC) adverse respiratory health effects compared to the regulated pollutants. We aimed to study the short-term associations between UFP and BC concentrations inside vehicles and (1) the onset of mucosal irritation and (2) the acute changes in lung function of Parisian taxi drivers during a working day. An epidemiological study was carried out on 50 taxi drivers in Paris. UFP and BC were measured inside their vehicles with DiSCmini® and microAeth®, respectively. On the same day, the frequency and the severity of nose, eye, and throat irritations were self-reported by each participant and a spirometry test was performed before and after the work shift. Multivariate analysis was used to evaluate the associations between in-taxis UFP and BC concentrations and mucosal irritation and lung function, after adjustment for potential confounders. In-taxis UFP concentrations ranged from 17.9 to 37.9 × 103 particles/cm³ and BC concentrations from 2.2 to 3.9 μg/m³, during a mean of 9 ± 2 working hours. Significant dose-response relationships were observed between in-taxis UFP concentrations and both nasal irritation and lung function. The increase of in-taxis UFP (for an interquartile range of 20 × 103 particles/cm3) was associated to an increase in nasal irritation (adjusted OR = 6.27 [95% CI: 1.02 to 38.62]) and to a reduction in forced expiratory flow at 25–75% by −7.44% [95% CI: −12.63 to −2.24], forced expiratory volume in one second by −4.46% [95% CI: −6.99 to −1.93] and forced vital capacity by −3.31% [95% CI: −5.82 to −0.80]. Such associations were not found with BC. Incident throat and eye irritations were not related to in-vehicle particles exposure; however, they were associated with outdoor air quality (estimated by the Atmo index) and in-vehicle humidity, respectively. This study is the first to show a significant association, within a short-period of time, between in-vehicle UFP exposure and acute respiratory effects in professional drivers.

Keywords: black carbon, lung function, mucosal irritation, taxi drivers, ultrafine particles

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5554 Payload Bay Berthing of an Underwater Vehicle With Vertically Actuated Thrusters

Authors: Zachary Cooper-Baldock, Paulo E. Santos, Russell S. A. Brinkworth, Karl Sammut

Abstract:

In recent years, large unmanned underwater vehicles such as the Boeing Voyager and Anduril Ghost Shark have been developed. These vessels can be structured to contain onboard internal payload bays. These payload bays can serve a variety of purposes – including the launch and recovery (LAR) of smaller underwater vehicles. The LAR of smaller vessels is extremely important, as it enables transportation over greater distances, increased time on station, data transmission and operational safety. The larger vessel and its payload bay structure complicate the LAR of UUVs in contrast to static docks that are affixed to the seafloor, as they actively impact the local flow field. These flow field impacts require analysis to determine if UUV vessels can be safely launched and recovered inside the motherships. This research seeks to determine the hydrodynamic forces exerted on a vertically over-actuated, small, unmanned underwater vehicle (OUUV) during an internal LAR manoeuvre and compare this to an under-actuated vessel (UUUV). In this manoeuvre, the OUUV is navigated through the stern wake region of the larger vessel to a set point within the internal payload bay. The manoeuvre is simulated using ANSYS Fluent computational fluid dynamics models, covering the entire recovery of the OUUV and UUUV. The analysis of the OUUV is compared against the UUUV to determine the differences in the exerted forces. Of particular interest are the drag, pressure, turbulence and flow field effects exerted as the OUUV is driven inside the payload bay of the larger vessel. The hydrodynamic forces and flow field disturbances are used to determine the feasibility of making such an approach. From the simulations, it was determined that there was no significant detrimental physical forces, particularly with regard to turbulence. The flow field effects exerted by the OUUV are significant. The vertical thrusters exert significant wake structures, but their orientation ensures the wake effects are exerted below the UUV, minimising the impact. It was also seen that OUUV experiences higher drag forces compared to the UUUV, which will correlate to an increased energy expenditure. This investigation found no key indicators that recovery via a mothership payload bay was not feasible. The turbulence, drag and pressure phenomenon were of a similar magnitude to existing static and towed dock structures.

Keywords: underwater vehicles, submarine, autonomous underwater vehicles, AUV, computational fluid dynamics, flow fields, pressure, turbulence, drag

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5553 3D Microscopy, Image Processing, and Analysis of Lymphangiogenesis in Biological Models

Authors: Thomas Louis, Irina Primac, Florent Morfoisse, Tania Durre, Silvia Blacher, Agnes Noel

Abstract:

In vitro and in vivo lymphangiogenesis assays are essential for the identification of potential lymphangiogenic agents and the screening of pharmacological inhibitors. In the present study, we analyse three biological models: in vitro lymphatic endothelial cell spheroids, in vivo ear sponge assay, and in vivo lymph node colonisation by tumour cells. These assays provide suitable 3D models to test pro- and anti-lymphangiogenic factors or drugs. 3D images were acquired by confocal laser scanning and light sheet fluorescence microscopy. Virtual scan microscopy followed by 3D reconstruction by image aligning methods was also used to obtain 3D images of whole large sponge and ganglion samples. 3D reconstruction, image segmentation, skeletonisation, and other image processing algorithms are described. Fixed and time-lapse imaging techniques are used to analyse lymphatic endothelial cell spheroids behaviour. The study of cell spatial distribution in spheroid models enables to detect interactions between cells and to identify invasion hierarchy and guidance patterns. Global measurements such as volume, length, and density of lymphatic vessels are measured in both in vivo models. Branching density and tortuosity evaluation are also proposed to determine structure complexity. Those properties combined with vessel spatial distribution are evaluated in order to determine lymphangiogenesis extent. Lymphatic endothelial cell invasion and lymphangiogenesis were evaluated under various experimental conditions. The comparison of these conditions enables to identify lymphangiogenic agents and to better comprehend their roles in the lymphangiogenesis process. The proposed methodology is validated by its application on the three presented models.

Keywords: 3D image segmentation, 3D image skeletonisation, cell invasion, confocal microscopy, ear sponges, light sheet microscopy, lymph nodes, lymphangiogenesis, spheroids

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5552 Potential Activities of Human Endogenous Retroviral kDNA in Melanoma Pathogenesis and HIV-1 Infection

Authors: Jianli Dong, Fangling Xu, Gengming Huang

Abstract:

Human endogenous retroviral elements (HERVs) comprise approximately 8% of the human genome. They are thought to be germline-integrated genetic remnants of retroviral infections. Although HERV sequences are highly defective, some, especially the K type (HERV-K), have been shown to be expressed and may have biological activities in the pathogenesis of cancer, chronic inflammation and autoimmune diseases. We found that HERV-K GAG and ENV proteins were strongly expressed in pleomorphic melanoma cells. We also detected a critical role of HERV-K ENV in mediating intercellular fusion and colony formation of melanoma cells. Interestingly, we found that levels of HERV-K GAG and ENV expression correlated with the activation of ERK and loss of p16INK4A in melanoma cells, and inhibition of MEK or CDK4, especially in combination, reduced HERV-K expression in melanoma cells. We also performed a reverse transcription-polymerase chain reaction (RT-PCR) assay using DNase I digestion to remove “contaminating” HERV-K genomic DNA and examined HERV-K RNA expression in plasma samples from HIV-1 infected individuals. We found a covariation between HERV-K RNA expression and CD4 cell counts in HIV-1 positive samples. Although a causal link between HERV-K activation and melanoma development, and between HERV-K activation, HIV-1 infection and CD4 cell count have yet to be determined, existing data support the further research efforts in HERV-K.

Keywords: CD4 cell, HERV-K, HIV-1, melanoma

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5551 Worst-Case Load Shedding in Electric Power Networks

Authors: Fu Lin

Abstract:

We consider the worst-case load-shedding problem in electric power networks where a number of transmission lines are to be taken out of service. The objective is to identify a prespecified number of line outages that lead to the maximum interruption of power generation and load at the transmission level, subject to the active power-flow model, the load and generation capacity of the buses, and the phase-angle limit across the transmission lines. For this nonlinear model with binary constraints, we show that all decision variables are separable except for the nonlinear power-flow equations. We develop an iterative decomposition algorithm, which converts the worst-case load shedding problem into a sequence of small subproblems. We show that the subproblems are either convex problems that can be solved efficiently or nonconvex problems that have closed-form solutions. Consequently, our approach is scalable for large networks. Furthermore, we prove the convergence of our algorithm to a critical point, and the objective value is guaranteed to decrease throughout the iterations. Numerical experiments with IEEE test cases demonstrate the effectiveness of the developed approach.

Keywords: load shedding, power system, proximal alternating linearization method, vulnerability analysis

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5550 Biodegradation Effects onto Source Identification of Diesel Fuel Contaminated Soils

Authors: Colin S. Chen, Chien-Jung Tien, Hsin-Jan Huang

Abstract:

For weathering studies, the change of chemical constituents by biodegradation effect in diesel-contaminated soils are important factors to be considered, especially when there is a prolonged period of weathering processes. The objective was to evaluate biodegradation effects onto hydrocarbon fingerprinting and distribution patterns of diesel fuels, fuel source screening and differentiation, source-specific marker compounds, and diagnostic ratios of diesel fuel constituents by laboratory and field studies. Biodegradation processes of diesel contaminated soils were evaluated by experiments lasting for 15 and 12 months, respectively. The degradation of diesel fuel in top soils was affected by organic carbon content and biomass of microorganisms in soils. Higher depletion of total petroleum hydrocarbon (TPH), n-alkanes, and polynuclear aromatic hydrocarbons (PAHs) and their alkyl homologues was observed in soils containing higher organic carbon content and biomass. Decreased ratio of selected isoprenoids (i.e., pristane (Pr) and phytane (Ph)) including n-C17/pristane and n-C18/phytane was observed. The ratio of pristane/phytane was remained consistent for a longer period of time. At the end of the experimental period, a decrease of pristane/phytane was observed. Biomarker compounds of bicyclic sesquiterpanes (BS) were less susceptible to the effects of biodegradation. The ratios of characteristic factors such as C15 sesquiterpane/ 8β(H)-drimane (BS3/BS5), C15 sesquiterpane/ 8β(H)-drimane (BS4/BS5), 8β(H)-drimane/8β(H)-homodrimane (BS5/BS10), and C15 sesquiterpane/8β(H)-homodrimane (BS3/BS10) could be adopted for source identification of diesel fuels in top soil. However, for biodegradation processes lasted for six months but shorter than nine months, only BS3/BS5 and BS3/BS10 could be distinguished in two diesel fuels. In subsoil experiments (contaminated soil located 50 cm below), the ratios of characteristic factors including BS3/BS5, BS4/BS5, and BS5/BS10 were valid for source identification of two diesel fuels for nine month biodegradation. At the early stage of contamination, biomass of soil decreased significantly. However, 6 and 7 dominant species were found in soils in top soil experiments, respectively. With less oxygen and nutrients in subsoil, less biomass of microorganisms was observed in subsoils. Only 2 and 4 diesel-degrading species of microorganisms were identified in two soils, respectively. Parameters of double ratio such as fluorene/C1-fluorene: C2-phenanthrene/C3-phenanthrene (C0F/C1F:C2P/C3P) in both top and subsoil, C2-naphthalene/C2-phenanthrene: C1-phenanthrene/C3-phenanthrene (C2N/C2P:C1P/C3P), and C1-phenanthrene/C1-fluorene: C3-naphthalene/C3-phenanthrene (C1P/C1F:C3N/C3P) in subsoil could serve as forensic indicators in diesel contaminated sites. BS3/BS10:BS4/BS5 could be used in 6 to 9 months of biodegradation processes. Results of principal component analysis (PCA) indicated that source identification of diesel fuels in top soil could only be perofrmed for weathering process less than 6 months. For subsoil, identification can be conducted for weathering process less than 9 months. Ratio of isoprenoids (pristane and phytane) and PAHs might be affected by biodegradation in spilled sites. The ratios of bicyclic sesquiterpanes could serve as forensic indicators in diesel-contaminated soils. Finally, source identification was attemped for samples collected from different fuel contaminated sites by using the unique pattern of sesquiterpanes. It was anticipated that the information generated from this study would be adopted by decision makers to evaluate the liability of cleanup in diesel contaminated sites.

Keywords: biodegradation, diagnostic ratio, diesel fuel, environmental forensics

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5549 Klotho Level as a Marker of Low Bone Mineral Density in Egyptian Sickle Cell Disease Patients

Authors: Mona Hamdy, Iman Shaheen, Hadeel Seif Eldin, Basma Ali, Omnia Abdeldayem

Abstract:

Summary: Bone involvement of sickle cell disease (SCD) patients varies from acute clinical manifestations of painful vaso-occlusive crises or osteomyelitis to more chronic affection of bone mineral density (BMD) and debilitating osteonecrosis and osteoporosis. Secreted klotho protein is involved in calcium (Ca) reabsorption in the kidney. This study aimed to measure serum klotho levels in children with SCD to determine the possibility of using it as a marker of low BMD in children with SCD in correlation with a dual-energy radiograph absorptiometry scan. This study included 60 sickle disease patients and 30 age-matched and sex-matched control participants without SCD. A highly statistically significant difference was found between patients with normal BMD and those with low BMD, with serum Ca and klotho levels being lower in the latter group. Klotho serum level correlated positively with both serum Ca and BMD. Serum klotho level showed 94.9% sensitivity and 95.2% specificity in the detection of low BMD. Both serum Ca and klotho serum levels may be useful markers for detection of low BMD related to SCD with high sensitivity and specificity; however, klotho may be a better indicator as it is less affected by the nutritional and endocrinal status of patients or by intake of Ca supplements.

Keywords: sickle cell disease, BMD, osteoporosis, DEXA, klotho

Procedia PDF Downloads 107
5548 Model the Off-Shore Ocean-Sea Waves to Generate Electric Power by Design of a Converting Device

Authors: Muthana A. M. Jameel Al-Jaboori

Abstract:

In this paper, we will present a mathematical model to design a system able to generate electricity from ocean-sea waves. We will use the basic principles of the transfer of the energy potential of waves in a chamber to force the air inside a vertical or inclined cylindrical column, which is topped by a wind turbine to rotate the electric generator. The present mathematical model included a high number of variables such as the wave, height, width, length, velocity, and frequency, as well as others for the energy cylindrical column, like varying diameters and heights, and the wave chamber shape diameter and height. While for the wells wind turbine the variables included the number of blades, length, width, and clearance, as well as the rotor and tip radius. Additionally, the turbine rotor and blades must be made from the light and strong material for a smooth blade surface. The variables were too vast and high in number. Then the program was run successfully within the MATLAB and presented very good modeling results.

Keywords: water wave, models, Wells turbine, MATLAB program

Procedia PDF Downloads 366
5547 Geoelectric Survey for Groundwater Potential in Waziri Umaru Federal Polytechnic, Birnin Kebbi, Nigeria

Authors: Ibrahim Mohammed, Suleiman Taofiq, Muhammad Naziru Yahya

Abstract:

Geoelectrical measurements using Schlumberger Vertical Electrical Sounding (VES) method were carried out in Waziri Umaru Federal Polytechnic, Birnin Kebbi, Nigeria, with the aim of determining the groundwater potential in the area. Twelve (12) Vertical Electric Sounding (VES) data were collected using Terrameter (ABEM SAS 300c) and analyzed using computer software (IPI2win), which gives an automatic interpretation of the apparent resistivity. The results of the interpretation of VES data were used in the characterization of three to five geo-electric layers from which the aquifer units were delineated. Data analysis indicated that water bearing formation exists in the third and fourth layers having resistivity range of 312 to 767 Ωm and 9.51 to 681 Ωm, respectively. The thickness of the formation ranges from 14.7 to 41.8 m, while the depth is from 8.22 to 53.7 m. Based on the result obtained from the interpretation of the data, five (5) VES stations were recommended as the most viable locations for groundwater exploration in the study area. The VES stations include VES A4, A5, A6, B1, and B2. The VES results of the entire area indicated that the water bearing formation occurs at maximum depth of 53.7 m at the time of this survey.

Keywords: aquifer, depth, groundwater, resistivity, Schlumberger

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5546 Cybernetic Model-Based Optimization of a Fed-Batch Process for High Cell Density Cultivation of E. Coli In Shake Flasks

Authors: Snehal D. Ganjave, Hardik Dodia, Avinash V. Sunder, Swati Madhu, Pramod P. Wangikar

Abstract:

Batch cultivation of recombinant bacteria in shake flasks results in low cell density due to nutrient depletion. Previous protocols on high cell density cultivation in shake flasks have relied mainly on controlled release mechanisms and extended cultivation protocols. In the present work, we report an optimized fed-batch process for high cell density cultivation of recombinant E. coli BL21(DE3) for protein production. A cybernetic model-based, multi-objective optimization strategy was implemented to obtain the optimum operating variables to achieve maximum biomass and minimized substrate feed rate. A syringe pump was used to feed a mixture of glycerol and yeast extract into the shake flask. Preliminary experiments were conducted with online monitoring of dissolved oxygen (DO) and offline measurements of biomass and glycerol to estimate the model parameters. Multi-objective optimization was performed to obtain the pareto front surface. The selected optimized recipe was tested for a range of proteins that show different extent soluble expression in E. coli. These included eYFP and LkADH, which are largely expressed in soluble fractions, CbFDH and GcanADH , which are partially soluble, and human PDGF, which forms inclusion bodies. The biomass concentrations achieved in 24 h were in the range 19.9-21.5 g/L, while the model predicted value was 19.44 g/L. The process was successfully reproduced in a standard laboratory shake flask without online monitoring of DO and pH. The optimized fed-batch process showed significant improvement in both the biomass and protein production of the tested recombinant proteins compared to batch cultivation. The proposed process will have significant implications in the routine cultivation of E. coli for various applications.

Keywords: cybernetic model, E. coli, high cell density cultivation, multi-objective optimization

Procedia PDF Downloads 263
5545 Biological Significance of Long Intergenic Noncoding RNA LINC00273 in Lung Cancer Cell Metastasis

Authors: Ipsita Biswas, Arnab Sarkar, Ashikur Rahaman, Gopeswar Mukherjee, Subhrangsu Chatterjee, Shamee Bhattacharjee, Deba Prasad Mandal

Abstract:

One of the major reasons for the high mortality rate of lung cancer is the substantial delays in disease detection at late metastatic stages. It is of utmost importance to understand the detailed molecular signaling and detect the molecular markers that can be used for the early diagnosis of cancer. Several studies explored the emerging roles of long noncoding RNAs (lncRNAs) in various cancers as well as lung cancer. A long non-coding RNA LINC00273 was recently discovered to promote cancer cell migration and invasion, and its positive correlation with the pathological stages of metastasis may prove it to be a potential target for inhibiting cancer cell metastasis. Comparing real-time expression of LINC00273 in various human clinical cancer tissue samples with normal tissue samples revealed significantly higher expression in cancer tissues. This long intergenic noncoding RNA was found to be highly expressed in human liver tumor-initiating cells, human gastric adenocarcinoma AGS cell line, as well as human non-small cell lung cancer A549 cell line. SiRNA and shRNA-induced knockdown of LINC00273 in both in vitro and in vivo nude mice significantly subsided AGS and A549 cancer cell migration and invasion. LINC00273 knockdown also reduced TGF-β induced SNAIL, SLUG, VIMENTIN, ZEB1 expression, and metastasis in A549 cells. Plenty of reports have suggested the role of microRNAs of the miR200 family in reversing epithelial to mesenchymal transition (EMT) by inhibiting ZEB transcription factors. In this study, hsa-miR-200a-3p was predicted via IntaRNA-Freiburg RNA tools to be a potential target of LINC00273 with a negative free binding energy of −8.793 kcal/mol, and this interaction was verified as a confirmed target of LINC00273 by RNA pulldown, real-time PCR and luciferase assay. Mechanistically, LINC00273 accelerated TGF-β induced EMT by sponging hsa-miR-200a-3p which in turn liberated ZEB1 and promoted prometastatic functions in A549 cells in vitro as verified by real-time PCR and western blotting. The similar expression patterns of these EMT regulatory pathway molecules, viz. LINC00273, hsa-miR-200a-3p, ZEB1 and TGF-β, were also detected in various clinical samples like breast cancer tissues, oral cancer tissues, lung cancer tissues, etc. Overall, this LINC00273 mediated EMT regulatory signaling can serve as a potential therapeutic target for the prevention of lung cancer metastasis.

Keywords: epithelial to mesenchymal transition, long noncoding RNA, microRNA, non-small-cell lung carcinoma

Procedia PDF Downloads 161
5544 Magnetic SF (Silk Fibroin) E-Gel Scaffolds Containing bFGF-Conjugated Fe3O4 Nanoparticles

Authors: Z. Karahaliloğlu, E. Yalçın, M. Demirbilek, E.B. Denkbaş

Abstract:

Critical-sized bone defects caused by trauma, bone diseases, prosthetic implant revision or tumor excision cannot be repaired by physiological regenerative processes. Current orthopedic applications for critical-sized bone defects are to use autologous bone grafts, bone allografts, or synthetic graft materials. However, these strategies are unable to solve completely the problem, and motivate the development of novel effective biological scaffolds for tissue engineering applications and regenerative medicine applications. In particular, scaffolds combined with a variety of bio-agents as fundamental tools emerge to provide the regeneration of damaged bone tissues due to their ability to promote cell growth and function. In this study, a magnetic silk fibroin (SF) hydrogel scaffold was prepared by electrogelation process of the concentrated Bombxy mori silk fibroin (8 %wt) aqueous solution. For enhancement of osteoblast-like cells (SaOS-2) growth and adhesion, basal fibroblast growth factor (bFGF) were conjugated physically to the HSA-coated magnetic nanoparticles (Fe3O4) and magnetic SF e-gel scaffolds were prepared by incorporation of Fe3O4, HSA (human serum albumin)=Fe3O4 and HSA=Fe3O4-bFGF nanoparticles. HSA=Fe3O4, HSA=Fe3O4-bFGF loaded and bare SF e-gels scaffolds were characterized using scanning electron microscopy (SEM.) For cell studies, human osteoblast-like cell line (SaOS-2) was used and an MTT assay was used to assess the cytotoxicity of magnetic silk fibroin e-gel scaffolds and cell density on these surfaces. For the evaluation osteogenic activation, ALP (alkaline phosphatase), the amount of mineralized calcium, total protein and collagen were studied. Fe3O4 nanoparticles were successfully synthesized and bFGF was conjugated to HSA=Fe3O4 nanoparticles with %97.5 of binding yield which has a particle size of 71.52±2.3 nm. Electron microscopy images of the prepared HSA and bFGF incorporated SF e-gel scaffolds showed a 3D porous morphology. In terms of water uptake results, bFGF conjugated HSA=Fe3O4 nanoparticles has the best water absorbability behavior among all groups. In the in-vitro cell culture studies realized using SaOS-2 cell line, the coating of Fe3O4 nanoparticles surface with a protein enhance the cell viability and HSA coating and bFGF conjugation, the both have an inductive effect in the cell proliferation. One of the markers of bone formation and osteoblast differentiation, according to the ALP activity and total protein results, HSA=Fe3O4-bFGF loaded SF e-gels had significantly enhanced ALP activity. Osteoblast cultured HSA=Fe3O4-bFGF loaded SF e-gels deposited more calcium compared with SF e-gel. The proposed magnetic scaffolds seem to be promising for bone tissue regeneration and used in future work for various applications.

Keywords: basic fibroblast growth factor (bFGF), e-gel, iron oxide nanoparticles, silk fibroin

Procedia PDF Downloads 294
5543 Life Cycle Assessment of Rare Earth Metals Production: Hotspot Analysis of Didymium Electrolysis Process

Authors: Sandra H. Fukurozaki, Andre L. N. Silva, Joao B. F. Neto, Fernando J. G. Landgraf

Abstract:

Nowadays, the rare earth (RE) metals play an important role in emerging technologies that are crucial for the decarbonisation of the energy sector. Their unique properties have led to increasing clean energy applications, such as wind turbine generators, and hybrid and electric vehicles. Despite the substantial media coverage that has recently surrounded the mining and processing of rare earth metals, very little quantitative information is available concerning their subsequent life stages, especially related to the metallic production of didymium (Nd-Pr) in fluoride molten salt system. Here we investigate a gate to gate scale life cycle assessment (LCA) of the didymium electrolysis based on three different scenarios of operational conditions. The product system is modeled with SimaPro Analyst 8.0.2 software, and IMPACT 2002+ was applied as an impact assessment tool. In order to develop a life cycle inventories built in software databases, patents, and other published sources together with energy/mass balance were utilized. Analysis indicates that from the 14 midpoint impact categories evaluated, the global warming potential (GWP) is the main contributors to the total environmental burden, ranging from 2.7E2 to 3.2E2 kg CO2eq/kg Nd-Pr. At the damage step assessment, the results suggest that slight changes in materials flows associated with enhancement of current efficiency (between 2.5% and 5%), could lead a reduction up to 12% and 15% of human health and climate change damage, respectively. Additionally, this paper highlights the knowledge gaps and future research efforts needing to understand the environmental impacts of Nd-Pr electrolysis process from the life cycle perspective.

Keywords: didymium electrolysis, environmental impacts, life cycle assessment, rare earth metals

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5542 Investigation the Photocatalytic Properties of Fe3O4-ZnO Nanocomposites Prepared by Sonochemical Method

Authors: Atena Naeimi, Mehri-Sadat Ekrami-Kakhki

Abstract:

Fe3O4 is one of the important magnetic oxides with spinel structure; it has exhibited unique electric and magnetic properties based on the electron transfer between Fe2+ and Fe3+ in the octahedral sites. Fe3O4 have received considerable attention in various areas such as cancer therapy, drug targeting, enzyme immobilization catalysis, magnetic cell separation, magnetic refrigeration systems and super-paramagnetic materials. Fe3O4–ZnO nanostructures were synthesized via a surfactant-free ultrasonic reaction at room temperatures. The effect of various parameters such as temperature, time, and power on the size and morphology of the product was investigated. Alternating gradient force magnetometer shows that Fe3O4 nanoparticles exhibit super-paramagnetic behaviour at room temperature. For preparation of nanocomposite 1 g of Fe3O4 nanostructures were dispersed in 100 mL of distilled water. 0.25 g of Zn (NO3)2 and 20 mL of NH3 solution 1 M were then slowly added to the solution under ultrasonic irradiation. The product was centrifuged, washed with distilled water and dried in the air. The photocatalytic behaviour of Fe3O4–ZnO nanoparticles was evaluated using the degradation of a methyl orange aqueous solution under ultraviolet light irradiation. As time increased, more and more methyl orange was adsorbed on the nanoparticles catalyst, until the absorption peak vanish. The methyl orange concentration decreased rapidly with increasing UV-irradiation time.

Keywords: nanocomposite, ultrasonic, paramagnetic, photocatalytic

Procedia PDF Downloads 304
5541 Improving the Bioprocess Phenotype of Chinese Hamster Ovary Cells Using CRISPR/Cas9 and Sponge Decoy Mediated MiRNA Knockdowns

Authors: Kevin Kellner, Nga Lao, Orla Coleman, Paula Meleady, Niall Barron

Abstract:

Chinese Hamster Ovary (CHO) cells are the prominent cell line used in biopharmaceutical production. To improve yields and find beneficial bioprocess phenotypes genetic engineering plays an essential role in recent research. The miR-23 cluster, specifically miR-24 and miR-27, was first identified as differentially expressed during hypothermic conditions suggesting a role in proliferation and productivity in CHO cells. In this study, we used sponge decoy technology to stably deplete the miRNA expression of the cluster. Furthermore, we implemented the CRISPR/Cas9 system to knockdown miRNA expression. Sponge constructs were designed for an imperfect binding of the miRNA target, protecting from RISC mediated cleavage. GuideRNAs for the CRISPR/Cas9 system were designed to target the seed region of the miRNA. The expression of mature miRNA and precursor were confirmed using RT-qPCR. For both approaches stable expressing mixed populations were generated and characterised in batch cultures. It was shown, that CRISPR/Cas9 can be implemented in CHO cells with achieving high knockdown efficacy of every single member of the cluster. Targeting of one miRNA member showed that its genomic paralog is successfully targeted as well. The stable depletion of miR-24 using CRISPR/Cas9 showed increased growth and specific productivity in a CHO-K1 mAb expressing cell line. This phenotype was further characterized using quantitative label-free LC-MS/MS showing 186 proteins differently expressed with 19 involved in proliferation and 26 involved in protein folding/translation. Targeting miR-27 in the same cell line showed increased viability in late stages of the culture compared to the control. To evaluate the phenotype in an industry relevant cell line; the miR-23 cluster, miR-24 and miR-27 were stably depleted in a Fc fusion CHO-S cell line which showed increased batch titers up to 1.5-fold. In this work, we highlighted that the stable depletion of the miR-23 cluster and its members can improve the bioprocess phenotype concerning growth and productivity in two different cell lines. Furthermore, we showed that using CRISPR/Cas9 is comparable to the traditional sponge decoy technology.

Keywords: Chinese Hamster ovary cells, CRISPR/Cas9, microRNAs, sponge decoy technology

Procedia PDF Downloads 203