Search results for: enzymatic fuel cell
4254 SIRT1 Gene Polymorphisms and Its Protein Level in Colorectal Cancer
Authors: Olfat Shaker, Miriam Wadie, Reham Ali, Ayman Yosry
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Colorectal cancer (CRC) is a major cause of mortality and morbidity and accounts for over 9% of cancer incidence worldwide. Silent information regulator 2 homolog 1 (SIRT1) gene is located in the nucleus and exert its effects via modulation of histone and non-histone targets. They function in the cell via histone deacetylase (HDAC) and/or adenosine diphosphate ribosyl transferase (ADPRT) enzymatic activity. The aim of this work was to study the relationship between SIRT1 polymorphism and its protein level in colorectal cancer patients in comparison to control cases. This study includes 2 groups: thirty healthy subjects (control group) & one hundred CRC patients. All subjects were subjected to: SIRT-1 serum level was measured by ELISA and gene polymorphisms of rs12778366, rs375891 and rs3740051 were detected by real time PCR. For CRC patients clinical data were collected (size, site of tumor as well as its grading, obesity) CRC patients showed high significant increase in the mean level of serum SIRT-1 compared to control group (P<0.001). Mean serum level of SIRT-1 showed high significant increase in patients with tumor size ≥5 compared to the size < 5 cm (P<0.05). In CRC patients, percentage of T allele of rs12778366 was significantly lower than controls, CC genotype and C allele C of rs 375891 were significantly higher than control group. In CRC patients, the CC genotype of rs12778366, was 75% in rectosigmoid and 25% in cecum & ascending colon. According to tumor size, the percentage of CC genotype was 87.5% in tumor size ≥5 cm. Conclusion: serum level of SIRT-1 and T allele, C allele of rs12778366 and rs 375891 respectively can be used as diagnostic markers for CRC patients.Keywords: CRC, SIRT1, polymorphisms, ELISA
Procedia PDF Downloads 2174253 Cloning, Expression and N-Terminal Pegylation of Human Interferon Alpha-2b Analogs and Their Cytotoxic Evaluation against Cancer Cell Lines
Authors: Syeda Kiran Shahzadi, Nasir Mahmood, Muhammad Abdul Qadir
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In the current research, three recombinant human interferon alpha-2b proteins (two modified and one normal form) were produced and Pegylated with an aim to produce more effective drugs against viral infections and cancers. The modified recombinant human interferon alpha-2b proteins were produced by site-directed modifications of interferon alpha 2b gene, targeting the amino acids at positions ‘R23’ and ‘H34’. The resulting chemically modified and unmodified forms of human interferon alpha 2b were conjugated with methoxy-polyethylene glycol propanealdehyde (400 KDa) and methoxy-polyethylene glycol succinimidyl succinate (400 KDa). Pegylation of normal and modified forms of Interferon alpha-2b prolong their release time and enhance their efficacy. The conjugation of PEG with modified and unmodified human interferon alpha 2b protein drugs was also characterized with 1H-NMR, HPLC, and SDS-PAGE. Antiproliferative assays of modified and unmodified forms of drugs were performed in cell based bioassays using MDBK cell lines. The results indicated that experimentally produced recombinant human interferon alpha-2b proteins were biologically active and resulted in significant inhibition of cell growth.Keywords: protein refolding, antiproliferative activities, biomedical applications, human interferon alpha-2b, pegylation, mPEG-propionaldehyde, site directed mutagenesis, E. coli expression
Procedia PDF Downloads 1754252 Experimental Study of Flow Effects of Solid Particles’ Size in Porous Media
Authors: S. Akridiss, E. El Tabach, K. Chetehouna, N. Gascoin, M. S. Kadiri
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Transpiration cooling combined to regenerative cooling is a technique that could be used to cool the porous walls of the future ramjet combustion chambers; it consists of using fuel that will flow through the pores of the porous material consisting of the chamber walls, as coolant. However, at high temperature, the fuel is pyrolysed and generates solid coke particles inside the porous materials. This phenomenon can lead to a significant decrease of the material permeability and can affect the efficiency of the cooling system. In order to better understand this phenomenon, an experimental laboratory study was undertaken to determine the transport and deposition of particles in a sintered porous material subjected to steady state flow. The test bench composed of a high-pressure autoclave is used to study the transport of different particle size (35Keywords: experimental study, permeability, porous material, suspended particles
Procedia PDF Downloads 2734251 Designing, Manufacturing and Testing a Portable Tractor Unit Biocoal Harvester Combine of Agriculture and Animal Wastes
Authors: Ali Moharrek, Hosein Mobli, Ali Jafari, Ahmad Tabataee Far
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Biomass is a material generally produced by plants living on soil or water and their derivatives. The remains of agricultural and forest products contain biomass which is changeable into fuel. Besides, you can obtain biogas and ethanol from the charcoal produced from biomass through specific actions. this technology was designed for as a useful Native Fuel and Technology in Energy disasters Management Due to the sudden interruption of the flow of heat energy One of the problems confronted by mankind in the future is the limitations of fossil energy which necessitates production of new energies such as biomass. In order to produce biomass from the remains of the plants, different methods shall be applied considering factors like cost of production, production technology, area of requirement, speed of work easy utilization, ect. In this article we are focusing on designing a biomass briquetting portable machine. The speed of installation of the machine on a tractor is estimated as 80 MF 258. Screw press is used in designing this machine. The needed power for running this machine which is estimated as 17.4 kW is provided by the power axis of tractor. The pressing speed of the machine is considered to be 375 RPM Finally the physical and mechanical properties of the product were compared with utilized material which resulted in appropriate outcomes. This machine is designed for Gathering Raw materials of the ground by Head Section. During delivering the raw materials to Briquetting section, they Crushed, Milled & Pre Heated in Transmission section. This machine is a Combine Portable Tractor unit machine and can use all type of Agriculture, Forest & Livestock Animals Resides as Raw material to make Bio fuel. The Briquetting Section was manufactured and it successfully made bio fuel of Sawdust. Also this machine made a biofuel with Ethanol of sugarcane Wastes. This Machine is using P.T.O power source for Briquetting and Hydraulic Power Source for Pre Processing of Row Materials.Keywords: biomass, briquette, screw press, sawdust, animal wastes, portable, tractors
Procedia PDF Downloads 3154250 Fermentation of Xylose and Glucose Mixture in Intensified Reactors by Scheffersomyces stipitis to Produce Ethanol
Authors: S. C. Santos, S. R. Dionísio, A. L. D. De Andrade, L. R. Roque, A. C. Da Costa, J. L. Ienczak
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In this work, two fermentations at different temperatures (25 and 30 ºC), with cell recycling, were accomplished to produce ethanol, using a mix of commercial substrates, xylose (70%) and glucose (30%), as organic source for Scheffersomyces stipitis. Five consecutive fermentations of 80 g L-1 (1º, 2º and 3º recycles), 96 g L-1 (4º recycle) and 120 g L-1 (5º recycle)reduced sugars led to a final maximum ethanol concentration of 17.2 and 34.5 g L-1, at 25 and 30 ºC, respectively. Glucose was the preferred substrate; moreover xylose startup degradation was initiated after a remaining glucose presence in the medium. Results showed that yeast acid treatment, performed before each cycle, provided improvements on cell viability, accompanied by ethanol productivity of 2.16 g L-1 h-1 at 30 ºC. A maximum 36% of xylose was retained in the fermentation medium and after five-cycle fermentation an ethanol yield of 0.43 g ethanol/g sugars was observed. S. stipitis fermentation capacity and tolerance showed better results at 30 ºC with 83.4% of theoretical yield referenced on initial biomass.Keywords: 5-carbon sugar, cell recycling fermenter, mixed sugars, xylose-fermenting yeast
Procedia PDF Downloads 4164249 Non-Burn Treatment of Health Care Risk Waste
Authors: Jefrey Pilusa, Tumisang Seodigeng
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This research discusses a South African case study for the potential of utilizing refuse-derived fuel (RDF) obtained from non-burn treatment of health care risk waste (HCRW) as potential feedstock for green energy production. This specific waste stream can be destroyed via non-burn treatment technology involving high-speed mechanical shredding followed by steam or chemical injection to disinfect the final product. The RDF obtained from this process is characterised by a low moisture, low ash, and high calorific value which means it can be potentially used as high-value solid fuel. Due to the raw feed of this RDF being classified as hazardous, the final RDF has been reported to be non-infectious and can blend with other combustible wastes such as rubber and plastic for waste to energy applications. This study evaluated non-burn treatment technology as a possible solution for on-site destruction of HCRW in South African private and public health care centres. Waste generation quantities were estimated based on the number of registered patient beds, theoretical bed occupancy. Time and motion study was conducted to evaluate the logistics viability of on-site treatment. Non-burn treatment technology for HCRW is a promising option for South Africa, and successful implementation of this method depends upon the initial capital investment, operational cost and environmental permitting of such technology; there are other influencing factors such as the size of the waste stream, product off-take price as well as product demand.Keywords: autoclave, disposal, fuel, incineration, medical waste
Procedia PDF Downloads 1764248 Study of the Biochemical Properties of the Protease Coagulant Milk Extracted from Sunflower Cake: Manufacturing Test of Cheeses Uncooked Dough Press and Analysis of Sensory Properties
Authors: Kahlouche Amal, Touzene F. Zohra, Betatache Fatihaet Nouani Abdelouahab
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The development of the world production of the cheese these last decades, as well as agents' greater request cheap coagulants, accentuated the search for new surrogates of the rennet. What about the interest to explore the vegetable biodiversity, the source well cheap of many naturals metabolites that the scientists today praise it (thistle, latex of fig tree, Cardoon, seeds of melon). Indeed, a big interest is concerned the search for surrogates of vegetable origin. The objective of the study is to show the possibility of extracting a protease coagulant the milk from the cake of Sunflower, available raw material and the potential source of surrogates of rennet. so, the determination of the proteolytic activity of raw extracts, the purification, the elimination of the pigments of tint of the enzymatic preparations, a better knowledge of the coagulative properties through study of the effect of certain factors (temperature, pH, concentration in CaCl2) are so many factors which contribute to value milk particularly those produced by the small ruminants of the Algerian dairy exploitations. Otherwise, extracts coagulants of vegetable origin allowed today to value traditional, in addition, although the extract coagulants of vegetable origin made it possible today to develop traditional cheeses whose Iberian peninsula is the promoter, but the test of 'pressed paste not cooked' cheese manufacturing led to the semi-scale pilot; and that, by using the enzymatic extract of sunflower (Helianthus annus) which gave satisfactory results as well to the level of outputs as on the sensory level,which, statistically,did not give any significant difference between studied cheeses. These results confirm the possibility of use of this coagulase as a substitute of rennet commercial on an industrial scale.Keywords: characterization, cheese, Rennet, sunflower
Procedia PDF Downloads 3504247 Computational Fluid Dynamics Design and Analysis of Aerodynamic Drag Reduction Devices for a Mazda T3500 Truck
Authors: Basil Nkosilathi Dube, Wilson R. Nyemba, Panashe Mandevu
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In highway driving, over 50 percent of the power produced by the engine is used to overcome aerodynamic drag, which is a force that opposes a body’s motion through the air. Aerodynamic drag and thus fuel consumption increase rapidly at speeds above 90kph. It is desirable to minimize fuel consumption. Aerodynamic drag reduction in highway driving is the best approach to minimize fuel consumption and to reduce the negative impacts of greenhouse gas emissions on the natural environment. Fuel economy is the ultimate concern of automotive development. This study aims to design and analyze drag-reducing devices for a Mazda T3500 truck, namely, the cab roof and rear (trailer tail) fairings. The aerodynamic effects of adding these append devices were subsequently investigated. To accomplish this, two 3D CAD models of the Mazda truck were designed using the Design Modeler. One, with these, append devices and the other without. The models were exported to ANSYS Fluent for computational fluid dynamics analysis, no wind tunnel tests were performed. A fine mesh with more than 10 million cells was applied in the discretization of the models. The realizable k-ε turbulence model with enhanced wall treatment was used to solve the Reynold’s Averaged Navier-Stokes (RANS) equation. In order to simulate the highway driving conditions, the tests were simulated with a speed of 100 km/h. The effects of these devices were also investigated for low-speed driving. The drag coefficients for both models were obtained from the numerical calculations. By adding the cab roof and rear (trailer tail) fairings, the simulations show a significant reduction in aerodynamic drag at a higher speed. The results show that the greatest drag reduction is obtained when both devices are used. Visuals from post-processing show that the rear fairing minimized the low-pressure region at the rear of the trailer when moving at highway speed. The rear fairing achieved this by streamlining the turbulent airflow, thereby delaying airflow separation. For lower speeds, there were no significant differences in drag coefficients for both models (original and modified). The results show that these devices can be adopted for improving the aerodynamic efficiency of the Mazda T3500 truck at highway speeds.Keywords: aerodynamic drag, computation fluid dynamics, fluent, fuel consumption
Procedia PDF Downloads 1364246 Power Supply by Soil Battery and Production of Hydrogen Fuel for Greenhouse and Space Heating
Authors: Mohsen Azarmjoo, Yasaman Azarmjoo, Zahra Alikhani Koopaei
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The increasing global population and continued growth in energy consumption underscore the need for renewable and sustainable energy sources more than ever. Soil batteries are a method for generating electrical energy by using recycled materials. Recycled materials include galvanized and copper sheets and recycled tires. Additionally, hydrogen, being a clean and efficient fuel, has the potential to replace fossil fuels. Consequently, hydrogen production from water presents a sustainable solution for energy supply. By utilizing aged materials, hydrogen production becomes more cost-effective and environmentally friendly. This article focuses on energy-deprived agricultural lands, explaining how soil batteries and hydrogen can provide the necessary energy for agricultural equipment, such as irrigation, lighting, greenhouse ventilation, and heating. The article explores the benefits of utilizing this method, emphasizing its potential to reduce environmental pollution through the use of recyclable materials. It is worth mentioning that these technologies face challenges, but their progress toward achieving zero-energy consumer standards positions them as promising future technologies for electricity generation. This article provides detailed insights into emerging technologies using a constructed case study involving soil batteries and a hydrogen fuel production device.Keywords: electricity generation, soil batteries, tires, hydrogen, heat supply, water, aged materials, recycling, agricultural lands
Procedia PDF Downloads 624245 Impact of Electric Vehicles on Energy Consumption and Environment
Authors: Amela Ajanovic, Reinhard Haas
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Electric vehicles (EVs) are considered as an important means to cope with current environmental problems in transport. However, their high capital costs and limited driving ranges state major barriers to a broader market penetration. The core objective of this paper is to investigate the future market prospects of various types of EVs from an economic and ecological point of view. Our method of approach is based on the calculation of total cost of ownership of EVs in comparison to conventional cars and a life-cycle approach to assess the environmental benignity. The most crucial parameters in this context are km driven per year, depreciation time of the car and interest rate. The analysis of future prospects it is based on technological learning regarding investment costs of batteries. The major results are the major disadvantages of battery electric vehicles (BEVs) are the high capital costs, mainly due to the battery, and a low driving range in comparison to conventional vehicles. These problems could be reduced with plug-in hybrids (PHEV) and range extenders (REXs). However, these technologies have lower CO₂ emissions in the whole energy supply chain than conventional vehicles, but unlike BEV they are not zero-emission vehicles at the point of use. The number of km driven has a higher impact on total mobility costs than the learning rate. Hence, the use of EVs as taxis and in car-sharing leads to the best economic performance. The most popular EVs are currently full hybrid EVs. They have only slightly higher costs and similar operating ranges as conventional vehicles. But since they are dependent on fossil fuels, they can only be seen as energy efficiency measure. However, they can serve as a bridging technology, as long as BEVs and fuel cell vehicle do not gain high popularity, and together with PHEVs and REX contribute to faster technological learning and reduction in battery costs. Regarding the promotion of EVs, the best results could be reached with a combination of monetary and non-monetary incentives, as in Norway for example. The major conclusion is that to harvest the full environmental benefits of EVs a very important aspect is the introduction of CO₂-based fuel taxes. This should ensure that the electricity for EVs is generated from renewable energy sources; otherwise, total CO₂ emissions are likely higher than those of conventional cars.Keywords: costs, mobility, policy, sustainability,
Procedia PDF Downloads 2244244 Enhancing Industrial Wastewater Treatment: Efficacy and Optimization of Ultrasound-Assisted Laccase Immobilized on Magnetic Fe₃O₄ Nanoparticles
Authors: K. Verma, v. S. Moholkar
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In developed countries, water pollution caused by industrial discharge has emerged as a significant environmental concern over the past decades. However, despite ongoing efforts, a fully effective and sustainable remediation strategy has yet to be identified. This paper describes how enzymatic and sonochemical treatments have demonstrated great promise in degrading bio-refractory pollutants. Mainly, a compelling area of interest lies in the combined technique of sono-enzymatic treatment, which has exhibited a synergistic enhancement effect surpassing that of the individual techniques. This study employed the covalent attachment method to immobilize Laccase from Trametes versicolor onto amino-functionalized magnetic Fe₃O₄ nanoparticles. To comprehensively characterize the synthesized free nanoparticles and the laccase-immobilized nanoparticles, various techniques such as X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FT-IR), scanning electron microscope (SEM), vibrating sample magnetometer (VSM), and surface area through Brunauer-Emmett-Teller (BET) were employed. The size of immobilized Fe₃O₄@Laccase was found to be 60 nm, and the maximum loading of laccase was found to be 24 mg/g of nanoparticle. An investigation was conducted to study the effect of various process parameters, such as immobilized Fe₃O₄ Laccase dose, temperature, and pH, on the % Chemical oxygen demand (COD) removal as a response. The statistical design pinpointed the optimum conditions (immobilized Fe₃O₄ Laccase dose = 1.46 g/L, pH = 4.5, and temperature = 66 oC), resulting in a remarkable 65.58% COD removal within 60 minutes. An even more significant improvement (90.31% COD removal) was achieved with ultrasound-assisted enzymatic reaction utilizing a 10% duty cycle. The investigation of various kinetic models for free and immobilized laccase, such as the Haldane, Yano, and Koga, and Michaelis-Menten, showed that ultrasound application impacted the kinetic parameters Vmax and Km. Specifically, Vmax values for free and immobilized laccase were found to be 0.021 mg/L min and 0.045 mg/L min, respectively, while Km values were 147.2 mg/L for free laccase and 136.46 mg/L for immobilized laccase. The lower Km and higher Vmax for immobilized laccase indicate its enhanced affinity towards the substrate, likely due to ultrasound-induced alterations in the enzyme's confirmation and increased exposure of active sites, leading to more efficient degradation. Furthermore, the toxicity and Liquid chromatography-mass spectrometry (LC-MS) analysis revealed that after the treatment process, the wastewater exhibited 70% less toxicity than before treatment, with over 25 compounds degrading by more than 75%. At last, the prepared immobilized laccase had excellent recyclability retaining 70% activity up to 6 consecutive cycles. A straightforward manufacturing strategy and outstanding performance make the recyclable magnetic immobilized Laccase (Fe₃O₄ Laccase) an up-and-coming option for various environmental applications, particularly in water pollution control and treatment.Keywords: kinetic, laccase enzyme, sonoenzymatic, ultrasound irradiation
Procedia PDF Downloads 664243 Development of a Plug-In Hybrid Powertrain System with Double Continuously Variable Transmissions
Authors: Cheng-Chi Yu, Chi-Shiun Chiou
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This study developed a plug-in hybrid powertrain system which consisted of two continuous variable transmissions. By matching between the engine, motor, generator, and dual continuous variable transmissions, this integrated power system can take advantages of the components. The hybrid vehicle can be driven by the internal combustion engine, or electric motor alone, or by these two power sources together when the vehicle is driven in hard acceleration or high load. The energy management of this integrated hybrid system controls the power systems based on rule-based control strategy to achieve better fuel economy. When the vehicle driving power demand is low, the internal combustion engine is operating in the low efficiency region, so the internal combustion engine is shut down, and the vehicle is driven by motor only. When the vehicle driving power demand is high, internal combustion engine would operate in the high efficiency region; then the vehicle could be driven by internal combustion engine. This strategy would operate internal combustion engine only in optimal efficiency region to improve the fuel economy. In this research, the vehicle simulation model was built in MATLAB/ Simulink environment. The analysis results showed that the power coupled efficiency of the hybrid powertrain system with dual continuous variable transmissions was better than that of the Honda hybrid system on the market.Keywords: plug-in hybrid power system, fuel economy, performance, continuously variable transmission
Procedia PDF Downloads 2874242 Sorghum Grains Grading for Food, Feed, and Fuel Using NIR Spectroscopy
Authors: Irsa Ejaz, Siyang He, Wei Li, Naiyue Hu, Chaochen Tang, Songbo Li, Meng Li, Boubacar Diallo, Guanghui Xie, Kang Yu
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Background: Near-infrared spectroscopy (NIR) is a non-destructive, fast, and low-cost method to measure the grain quality of different cereals. Previously reported NIR model calibrations using the whole grain spectra had moderate accuracy. Improved predictions are achievable by using the spectra of whole grains, when compared with the use of spectra collected from the flour samples. However, the feasibility for determining the critical biochemicals, related to the classifications for food, feed, and fuel products are not adequately investigated. Objectives: To evaluate the feasibility of using NIRS and the influence of four sample types (whole grains, flours, hulled grain flours, and hull-less grain flours) on the prediction of chemical components to improve the grain sorting efficiency for human food, animal feed, and biofuel. Methods: NIR was applied in this study to determine the eight biochemicals in four types of sorghum samples: hulled grain flours, hull-less grain flours, whole grains, and grain flours. A total of 20 hybrids of sorghum grains were selected from the two locations in China. Followed by NIR spectral and wet-chemically measured biochemical data, partial least squares regression (PLSR) was used to construct the prediction models. Results: The results showed that sorghum grain morphology and sample format affected the prediction of biochemicals. Using NIR data of grain flours generally improved the prediction compared with the use of NIR data of whole grains. In addition, using the spectra of whole grains enabled comparable predictions, which are recommended when a non-destructive and rapid analysis is required. Compared with the hulled grain flours, hull-less grain flours allowed for improved predictions for tannin, cellulose, and hemicellulose using NIR data. Conclusion: The established PLSR models could enable food, feed, and fuel producers to efficiently evaluate a large number of samples by predicting the required biochemical components in sorghum grains without destruction.Keywords: FT-NIR, sorghum grains, biochemical composition, food, feed, fuel, PLSR
Procedia PDF Downloads 664241 RACK1 Integrates Light and Brassinosteroid Signaling to Coordinate Cell Division During Root Soil Penetration
Authors: Liang Jiansheng, Zhu Wei
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Light and brassinosteroids are essential external and internal cues for plant survival. Although the coordination of light with phytohormone signals is crucial for plant growth and development, the molecular connection between light and brassinosteroid signaling during root soil penetration remains elusive. Here, we reveal that light-stabilized RACK1 couples a brassinosteroid signaling cascade to drive cell division in root meristems. RACK1 family scaffold proteins positively regulate light-induced the promotion of root elongation during soil penetration. Under the light condition, RACK1A interacts with both phyB and SPA1, then reinforces the phyB-SPA1 association to accumulate its abundance in roots. In response to brassinosteroid signals, RACK1A competes with BKI1 to attenuate the BRI1-BKI1 interaction, thereby leading to activating BRI1 actions in root development. Furthermore, RACK1A binds to BES1 to repress its DNA binding activity toward the target gene CYCD3;1. This ultimately allows to release the inhibition of CYCD3;1 transcription, and promotes cell division during root growth. Our study illustrates a new mechanistic model of how plants engage scaffold proteins in transducing light information to facilitate brassinosteroid signaling for root growth in the soil.Keywords: root growth, cell division, light signaling, brassinosteroid signaling, soil penetration, scaffold protein, RACK1
Procedia PDF Downloads 774240 Study into the Interactions of Primary Limbal Epithelial Stem Cells and HTCEPI Using Tissue Engineered Cornea
Authors: Masoud Sakhinia, Sajjad Ahmad
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Introduction: Though knowledge of the compositional makeup and structure of the limbal niche has progressed exponentially during the past decade, much is yet to be understood. Identifying the precise profile and role of the stromal makeup which spans the ocular surface may inform researchers of the most optimum conditions needed to effectively expand LESCs in vitro, whilst preserving their differentiation status and phenotype. Limbal fibroblasts, as opposed to corneal fibroblasts are thought to form an important component of the microenvironment where LESCs reside. Methods: The corneal stroma was tissue engineered in vitro using both limbal and corneal fibroblasts embedded within a tissue engineered 3D collagen matrix. The effect of these two different fibroblasts on LESCs and hTCEpi corneal epithelial cell line were then subsequently determined using phase contrast microscopy, histolological analysis and PCR for specific stem cell markers. The study aimed to develop an in vitro model which could be used to determine whether limbal, as opposed to corneal fibroblasts, maintained the stem cell phenotype of LESCs and hTCEpi cell line. Results: Tissue culture analysis was inconclusive and required further quantitative analysis for remarks on cell proliferation within the varying stroma. Histological analysis of the tissue-engineered cornea showed a comparable structure to that of the human cornea, though with limited epithelial stratification. PCR results for epithelial cell markers of cells cultured on limbal fibroblasts showed reduced expression of CK3, a negative marker for LESC’s, whilst also exhibiting a relatively low expression level of P63, a marker for undifferentiated LESCs. Conclusion: We have shown the potential for the construction of a tissue engineered human cornea using a 3D collagen matrix and described some preliminary results in the analysis of the effects of varying stroma consisting of limbal and corneal fibroblasts, respectively, on the proliferation of stem cell phenotype of primary LESCs and hTCEpi corneal epithelial cells. Although no definitive marker exists to conclusively illustrate the presence of LESCs, the combination of positive and negative stem cell markers in our study were inconclusive. Though it is less traslational to the human corneal model, the use of conditioned medium from that of limbal and corneal fibroblasts may provide a more simple avenue. Moreover, combinations of extracellular matrices could be used as a surrogate in these culture models.Keywords: cornea, Limbal Stem Cells, tissue engineering, PCR
Procedia PDF Downloads 2764239 Evaluation of the Cytotoxicity and Cellular Uptake of a Cyclodextrin-Based Drug Delivery System for Cancer Therapy
Authors: Caroline Mendes, Mary McNamara, Orla Howe
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Drug delivery systems are proposed for use in cancer treatment to specifically target cancer cells and deliver a therapeutic dose without affecting normal cells. For that purpose, the use of folate receptors (FR) can be considered a key strategy, since they are commonly over-expressed in cancer cells. In this study, cyclodextrins (CD) have being used as vehicles to target FR and deliver the chemotherapeutic drug, methotrexate (MTX). CDs have the ability to form inclusion complexes, in which molecules of suitable dimensions are included within their cavities. Here, β-CD has been modified using folic acid so as to specifically target the FR. Thus, this drug delivery system consists of β-CD, folic acid and MTX (CDEnFA:MTX). Cellular uptake of folic acid is mediated with high affinity by folate receptors while the cellular uptake of antifolates, such as MTX, is mediated with high affinity by the reduced folate carriers (RFCs). This study addresses the gene (mRNA) and protein expression levels of FRs and RFCs in the cancer cell lines CaCo-2, SKOV-3, HeLa, MCF-7, A549 and the normal cell line BEAS-2B, quantified by real-time polymerase chain reaction (real-time PCR) and flow cytometry, respectively. From that, four cell lines with different levels of FRs, were chosen for cytotoxicity assays of MTX and CDEnFA:MTX using the MTT assay. Real-time PCR and flow cytometry data demonstrated that all cell lines ubiquitously express moderate levels of RFC. These experiments have also shown that levels of FR protein in CaCo-2 cells are high, while levels in SKOV-3, HeLa and MCF-7 cells are moderate. A549 and BEAS-2B cells express low levels of FR protein. FRs are highly expressed in all the cancer cell lines analysed when compared to the normal cell line BEAS-2B. The cell lines CaCo-2, MCF-7, A549 and BEAS-2B were used in the cell viability assays. 48 hours treatment with the free drug and the complex resulted in IC50 values of 93.9 µM ± 15.2 and 56.0 µM ± 4.0 for CaCo-2 for free MTX and CDEnFA:MTX respectively, 118.2 µM ± 16.8 and 97.8 µM ± 12.3 for MCF-7, 36.4 µM ± 6.9 and 75.0 µM ± 10.5 for A549 and 132.6 µM ± 16.1 and 288.1 µM ± 26.3 for BEAS-2B. These results demonstrate that free MTX is more toxic towards cell lines expressing low levels of FR, such as the BEAS-2B. More importantly, these results demonstrate that the inclusion complex CDEnFA:MTX showed greater cytotoxicity than the free drug towards the high FR expressing CaCo-2 cells, indicating that it has potential to target this receptor, enhancing the specificity and the efficiency of the drug. The use of cell imaging by confocal microscopy has allowed visualisation of FR targeting in cancer cells, as well as the identification of the interlisation pathway of the drug. Hence, the cellular uptake and internalisation process of this drug delivery system is being addressed.Keywords: cancer treatment, cyclodextrins, drug delivery, folate receptors, reduced folate carriers
Procedia PDF Downloads 3094238 Tumor Cell Detection, Isolation and Monitoring Using Bi-Layer Magnetic Microfluidic Chip
Authors: Amir Seyfoori, Ehsan Samiei, Mohsen Akbari
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The use of microtechnology for detection and high yield isolation of circulating tumor cells (CTCs) has shown enormous promise as an indication of clinical metastasis prognosis and cancer treatment monitoring. The Immunomagnetic assay has been also coupled to microtechnology to improve the selectivity and efficiency of the current methods of cancer biomarker isolation. In this way, generation and configuration of the local high gradient magnetic field play essential roles in such assay. Additionally, considering the intrinsic heterogeneity of cancer cells, real-time analysis of isolated cells is necessary to characterize their responses to therapy. Totally, on-chip isolation and monitoring of the specific tumor cells is considered as a pressing need in the way of modified cancer therapy. To address these challenges, we have developed a bi-layer magnetic-based microfluidic chip for enhanced CTC detection and capturing. Micromagnet arrays at the bottom layer of the chip were fabricated using a new method of magnetic nanoparticle paste deposition so that they were arranged at the center of the chain microchannel with the lowest fluid velocity zone. Breast cancer cells labelled with EPCAM-conjugated smart microgels were immobilized on the tip of the micromagnets with greater localized magnetic field and stronger cell-micromagnet interaction. Considering different magnetic nano-powder usage (MnFe2O4 & gamma-Fe2O3) and micromagnet shapes (ellipsoidal & arrow), the capture efficiency of the systems was adjusted while the higher CTC capture efficiency was acquired for MnFe2O4 arrow micromagnet as around 95.5%. As a proof of concept of on-chip tumor cell monitoring, magnetic smart microgels made of thermo-responsive poly N-isopropylacrylamide-co-acrylic acid (PNIPAM-AA) composition were used for both purposes of targeted cell capturing as well as cell monitoring using antibody conjugation and fluorescent dye loading at the same time. In this regard, magnetic microgels were successfully used as cell tracker after isolation process so that by raising the temperature up to 37⁰ C, they released the contained dye and stained the targeted cell just after capturing. This microfluidic device was able to provide a platform for detection, isolation and efficient real-time analysis of specific CTCs in the liquid biopsy of breast cancer patients.Keywords: circulating tumor cells, microfluidic, immunomagnetic, cell isolation
Procedia PDF Downloads 1424237 Impact of Wheel-Housing on Aerodynamic Drag and Effect on Energy Consumption on an Bus
Authors: Amitabh Das, Yash Jain, Mohammad Rafiq B. Agrewale, K. C. Vora
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Role of wheel and underbody aerodynamics of vehicle in the formation of drag forces is detrimental to the fuel (energy) consumption during the course of operation at high velocities. This paper deals with the CFD simulation of the flow around the wheels of a bus with different wheel housing geometry and pattern. Based on benchmarking a model of a bus is selected and analysis is performed. The aerodynamic drag coefficient is obtained and turbulence around wheels is observed using ANSYS Fluent CFD simulation for different combinations of wheel-housing at the front wheels, at the rear wheels and both in the front and rear wheels. The drag force is recorded and corresponding influence on energy consumption on an electric bus is evaluated mathematically. A comparison is drawn between energy consumption of bus body without wheel housing and bus body with wheel housing. The result shows a significant reduction in drag coefficient and fuel consumption.Keywords: wheel-housing, CFD simulation, drag coefficient, energy consumption
Procedia PDF Downloads 1824236 Simulation Research of Diesel Aircraft Engine
Authors: Łukasz Grabowski, Michał Gęca, Mirosław Wendeker
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This paper presents the simulation results of a new opposed piston diesel engine to power a light aircraft. Created in the AVL Boost, the model covers the entire charge passage, from the inlet up to the outlet. The model shows fuel injection into cylinders and combustion in cylinders. The calculation uses the module for two-stroke engines. The model was created using sub-models available in this software that structure the model. Each of the sub-models is complemented with parameters in line with the design premise. Since engine weight resulting from geometric dimensions is fundamental in aircraft engines, two configurations of stroke were studied. For each of the values, there were calculated selected operating conditions defined by crankshaft speed. The required power was achieved by changing air fuel ratio (AFR). There was also studied brake specific fuel consumption (BSFC). For stroke S1, the BSFC was lowest at all of the three operating points. This difference is approximately 1-2%, which means higher overall engine efficiency but the amount of fuel injected into cylinders is larger by several mg for S1. The cylinder maximum pressure is lower for S2 due to the fact that compressor gear driving remained the same and boost pressure was identical in the both cases. Calculations for various values of boost pressure were the next stage of the study. In each of the calculation case, the amount of fuel was changed to achieve the required engine power. In the former case, the intake system dimensions were modified, i.e. the duct connecting the compressor and the air cooler, so its diameter D = 40 mm was equal to the diameter of the compressor outlet duct. The impact of duct length was also examined to be able to reduce the flow pulsation during the operating cycle. For the so selected geometry of the intake system, there were calculations for various values of boost pressure. The boost pressure was changed by modifying the gear driving the compressor. To reach the required level of cruising power N = 68 kW. Due to the mechanical power consumed by the compressor, high pressure ratio results in a worsened overall engine efficiency. The figure on the change in BSFC from 210 g/kWh to nearly 270 g/kWh shows this correlation and the overall engine efficiency is reduced by about 8%. Acknowledgement: This work has been realized in the cooperation with The Construction Office of WSK "PZL-KALISZ" S.A." and is part of Grant Agreement No. POIR.01.02.00-00-0002/15 financed by the Polish National Centre for Research and Development.Keywords: aircraft, diesel, engine, simulation
Procedia PDF Downloads 2064235 Effect of Injection Pressure and Fuel Injection Timing on Emission and Performance Characteristics of Karanja Biodiesel and its Blends in CI Engine
Authors: Mohan H., C. Elajchet Senni
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In the present of high energy consumption in every sphere of life, renewable energy sources are emerging as alternative to conventional fuels for energy security, mitigating green house gas emission and climate change. There has been a world wide interest in searching for alternatives to petroleum derived fuels due to their depletion as well as due to the concern for the environment. Vegetable oils have capability to solve this problem because they are renewable and lead to reduction in environmental pollution. But high smoke emission and lower thermal efficiency are the main problems associated with the use of neat vegetable oils in diesel engines. In the present work, performance, combustion and emission characteristics of CI engine fuelled with 20% by vol. methyl esters mixed with Karanja seed Oil, and Fuel injection pressures of 200 bar and 240 bar, injection timings (21°,23° and 25° BTDC) and Proportion B20 diesel respectively. Vegetable oils have capability to solve this problem because they are renewable and lead to reduction in environmental pollution. But, high smoke emission and lower thermal efficiency are the main problems associated with the use of neat vegetable oils in diesel engines. In the present work, performance, combustion and emission characteristics of CI engine fuelled with 20% by vol. methyl esters mixed with Karanja seed Oil, and Fuel injection pressures of 200 bar and 240 bar ,Injection timings (21°,23° and 25° BTDC) and Proportion B20 diesel respectively. Various performance, combustion and emission characteristics such as thermal efficiency, and brake specific fuel consumption, maximum cylinder pressure, instantaneous heat release, cumulative heat release with respect to crank angle, ignition lag, combustion duration, HC, NOx, CO, exhaust temperature and smoke intensity were measured.Keywords: karanja oil, injection pressure, injection timing, karanja oil methyl ester
Procedia PDF Downloads 2894234 Statistical Analysis of Parameters Effects on Maximum Strain and Torsion Angle of FRP Honeycomb Sandwich Panels Subjected to Torsion
Authors: Mehdi Modabberifar, Milad Roodi, Ehsan Souri
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In recent years, honeycomb fiber reinforced plastic (FRP) sandwich panels have been increasingly used in various industries. Low weight, low price, and high mechanical strength are the benefits of these structures. However, their mechanical properties and behavior have not been fully explored. The objective of this study is to conduct a combined numerical-statistical investigation of honeycomb FRP sandwich beams subject to torsion load. In this paper, the effect of geometric parameters of the sandwich panel on the maximum shear strain in both face and core and angle of torsion in a honeycomb FRP sandwich structures in torsion is investigated. The effect of Parameters including core thickness, face skin thickness, cell shape, cell size, and cell thickness on mechanical behavior of the structure were numerically investigated. Main effects of factors were considered in this paper and regression equations were derived. Taguchi method was employed as experimental design and an optimum parameter combination for the maximum structure stiffness has been obtained. The results showed that cell size and face skin thickness have the most significant impacts on torsion angle, maximum shear strain in face and core.Keywords: finite element, honeycomb FRP sandwich panel, torsion, civil engineering
Procedia PDF Downloads 4164233 Redox-labeled Electrochemical Aptasensor Array for Single-cell Detection
Authors: Shuo Li, Yannick Coffinier, Chann Lagadec, Fabrizio Cleri, Katsuhiko Nishiguchi, Akira Fujiwara, Soo Hyeon Kim, Nicolas Clément
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The need for single cell detection and analysis techniques has increased in the past decades because of the heterogeneity of individual living cells, which increases the complexity of the pathogenesis of malignant tumors. In the search for early cancer detection, high-precision medicine and therapy, the technologies most used today for sensitive detection of target analytes and monitoring the variation of these species are mainly including two types. One is based on the identification of molecular differences at the single-cell level, such as flow cytometry, fluorescence-activated cell sorting, next generation proteomics, lipidomic studies, another is based on capturing or detecting single tumor cells from fresh or fixed primary tumors and metastatic tissues, and rare circulating tumors cells (CTCs) from blood or bone marrow, for example, dielectrophoresis technique, microfluidic based microposts chip, electrochemical (EC) approach. Compared to other methods, EC sensors have the merits of easy operation, high sensitivity, and portability. However, despite various demonstrations of low limits of detection (LOD), including aptamer sensors, arrayed EC sensors for detecting single-cell have not been demonstrated. In this work, a new technique based on 20-nm-thick nanopillars array to support cells and keep them at ideal recognition distance for redox-labeled aptamers grafted on the surface. The key advantages of this technology are not only to suppress the false positive signal arising from the pressure exerted by all (including non-target) cells pushing on the aptamers by downward force but also to stabilize the aptamer at the ideal hairpin configuration thanks to a confinement effect. With the first implementation of this technique, a LOD of 13 cells (with5.4 μL of cell suspension) was estimated. In further, the nanosupported cell technology using redox-labeled aptasensors has been pushed forward and fully integrated into a single-cell electrochemical aptasensor array. To reach this goal, the LOD has been reduced by more than one order of magnitude by suppressing parasitic capacitive electrochemical signals by minimizing the sensor area and localizing the cells. Statistical analysis at the single-cell level is demonstrated for the recognition of cancer cells. The future of this technology is discussed, and the potential for scaling over millions of electrodes, thus pushing further integration at sub-cellular level, is highlighted. Despite several demonstrations of electrochemical devices with LOD of 1 cell/mL, the implementation of single-cell bioelectrochemical sensor arrays has remained elusive due to their challenging implementation at a large scale. Here, the introduced nanopillar array technology combined with redox-labeled aptamers targeting epithelial cell adhesion molecule (EpCAM) is perfectly suited for such implementation. Combining nanopillar arrays with microwells determined for single cell trapping directly on the sensor surface, single target cells are successfully detected and analyzed. This first implementation of a single-cell electrochemical aptasensor array based on Brownian-fluctuating redox species opens new opportunities for large-scale implementation and statistical analysis of early cancer diagnosis and cancer therapy in clinical settings.Keywords: bioelectrochemistry, aptasensors, single-cell, nanopillars
Procedia PDF Downloads 1154232 Flowsheet Development, Simulation and Optimization of Carbon-Di-Oxide Removal System at Natural Gas Reserves by Aspen–Hysys Process Simulator
Authors: Mohammad Ruhul Amin, Nusrat Jahan
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Natural gas is a cleaner fuel compared to the others. But it needs some treatment before it is in a state to be used. So natural gas purification is an integral part of any process where natural gas is used as raw material or fuel. There are several impurities in natural gas that have to be removed before use. CO2 is one of the major contaminants. In this project we have removed CO2 by amine process by using MEA solution. We have built up the whole amine process for removing CO2 in Aspen Hysys and simulated the process. At the end of simulation we have got very satisfactory results by using MEA solution for the removal of CO2. Simulation result shows that amine absorption process enables to reduce CO2 content from NG by 58%. HYSYS optimizer allowed us to get a perfect optimized plant. After optimization the profit of existing plant is increased by 2.34 %.Simulation and optimization by Aspen-HYSYS simulator makes available us to enormous information which will help us to further research in future.Keywords: Aspen–Hysys, CO2 removal, flowsheet development, MEA solution, natural gas optimization
Procedia PDF Downloads 4974231 Modeling of a Concentrating Photovoltaic Module with and without Cooling System
Authors: Intissar Benrhouma, Marta Victoria, Ignacio Anton, Bechir Chaouachi
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Concentrating photovoltaic systems CPV use optical elements, such as Fresnel lenses, to concentrate solar intensity. The concentrated solar energy is delivered to the solar cell from 20 to 100 W/cm². Some of this energy is converted to electricity, while the rest must be disposed of as a residual heat. Solar cells cooling should be a necessary part of CPV modeling because these systems allowed increasing the power received by the cell. This high power can rise the electrons’ potential causing the heating of the cell, which reduces the global module’s efficiency. This work consists of modeling a concentrating photovoltaic module with and without a cooling system. We have established a theoretical model based on energy balances carried out on a photovoltaic module using solar radiation concentration cells. Subsequently, we developed a calculation program on Matlab which allowed us to simulate the functioning of this module. The obtained results show that the addition of a cooling system to the module improves greatly the performance of our CPV system.Keywords: solar energy, photovoltaic, concentration, cooling, performance improvement
Procedia PDF Downloads 3964230 Evaluation of Iron Oxide-Functionalized Multiwall Carbon Nanotube Self-Standing Electrode for Symmetric Supercapacitor Application
Authors: B. V. Bhaskara Rao, Rodrigo Espinoza
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The rapid development of renewable energy sources has drawn great attention to energy storage devices, especially supercapacitors, because of their high power density and rate performance. This work focus on Fe₃O₄ nanoparticles synthesized by reverse co-precipitation and MWCNTs functionalized by –COOH acid functionalization. The results show that Optimized 25wt% Fe₃O₄@FMWCNT show high specific capacitance 100 mF/cm² at one mA/cm² whereas 15wt% Fe₃O₄@FMWCNT showed high stability (80% retention capacity) over 5000 cycles. The electrolyte used in the coin cell is LiPF6 and the thickness of the electrode is 30 microns. The optimized Fe₃O₄@FMWCNT bucky papers coin cell electrochemical studies suggest that 25wt% Fe₃O₄@FMWCNT could be a good candidate for high-capacity supercapacitor devices. This could be further tested for flexible and planar supercapacitor device application with gel electrolytes.Keywords: self-standing electrode, Fe₃O4@FMWCNT, supercapacitor, symmetric coin-cell
Procedia PDF Downloads 1544229 Binding Mechanism of Synthesized 5β-Dihydrocortisol and 5β-Dihydrocortisol Acetate with Human Serum Albumin to Understand Their Role in Breast Cancer
Authors: Monika Kallubai, Shreya Dubey, Rajagopal Subramanyam
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Our study is all about the biological interactions of synthesized 5β-dihydrocortisol (Dhc) and 5β-dihydrocortisol acetate (DhcA) molecules with carrier protein Human Serum Albumin (HSA). The cytotoxic study was performed on breast cancer cell line (MCF-7) normal human embryonic kidney cell line (HEK293), the IC50 values for MCF-7 cells were 28 and 25 µM, respectively, whereas no toxicity in terms of cell viability was observed with HEK293 cell line. The further experiment proved that Dhc and DhcA induced 35.6% and 37.7% early apoptotic cells and 2.5%, 2.9% late apoptotic cells respectively. Morphological observation of cell death through TUNEL assay revealed that Dhc and DhcA induced apoptosis in MCF-7 cells. The complexes of HSA–Dhc and HSA–DhcA were observed as static quenching, and the binding constants (K) was 4.7±0.03×104 M-1 and 3.9±0.05×104 M-1, and their binding free energies were found to be -6.4 and -6.16 kcal/mol, respectively. The displacement studies confirmed that lidocaine 1.4±0.05×104 M-1 replaced Dhc, and phenylbutazone 1.5±0.05×104 M-1 replaced by DhcA, which explains domain I and domain II are the binding sites for Dhc and DhcA. Further, CD results revealed that the secondary structure of HSA was altered in the presence of Dhc and DhcA. Furthermore, the atomic force microscopy and transmission electron microscopy showed that the dimensions like height and molecular sizes of the HSA–Dhc and HSA–DhcA complex were larger compared to HSA alone. Detailed analysis through molecular dynamics simulations also supported the greater stability of HSA–Dhc and HSA–DhcA complexes, and root-mean-square-fluctuation interpreted the binding site of Dhc as domain IB and domain IIA for DhcA. This information is valuable for the further development of steroid derivatives with improved pharmacological significance as novel anti-cancer drugs.Keywords: apoptosis, dihydrocortisol, fluorescence quenching, protein conformations
Procedia PDF Downloads 1294228 An Industrial Scada System Remote Control Using Mobile Phones
Authors: Ahmidah Elgali
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SCADA is the abbreviation for "Administrative Control And Data Acquisition." SCADA frameworks are generally utilized in industry for administrative control and information securing of modern cycles. Regular SCADA frameworks use PC, journal, slim client, and PDA as a client. In this paper, a Java-empowered cell phone has been utilized as a client in an example SCADA application to show and regulate the place of an example model crane. The paper presents a genuine execution of the online controlling of the model crane through a cell phone. The remote correspondence between the cell phone and the SCADA server is performed through a base station by means of general parcel radio assistance GPRS and remote application convention WAP. This application can be used in industrial sites in areas that are likely to be exposed to a security emergency (like terrorist attacks) which causes the sudden exit of the operators; however, no time to perform the shutdown procedures for the plant. Hence this application allows shutting down units and equipment remotely by mobile and so avoids damage and losses.Keywords: control, industrial, mobile, network, remote, SCADA
Procedia PDF Downloads 764227 The Impact of the Cell-Free Solution of Lactic Acid Bacteria on Cadaverine Production by Listeria monocytogenes and Staphylococcus aureus in Lysine-Decarboxylase Broth
Authors: Fatih Özogul, Nurten Toy, Yesim Özogul
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The influences of cell-free solutions (CFSs) of lactic acid bacteria (LAB) on cadaverine and other biogenic amine production by Listeria monocytogenes and Staphylococcus aureus were investigated in lysine decarboxylase broth (LDB) using HPLC. Cell-free solutions were prepared from Lactococcus lactis subsp. lactis, Leuconostoc mesenteroides subsp. cremoris, Pediococcus acidophilus and Streptococcus thermophiles. Two different concentrations that were 50% and 25% CFS and the control without CFSs were prepared. Significant variations on biogenic amine production were observed in the presence of L. monocytogenes and S. aureus (P<0.05). The role of CFS on biogenic amine production by foodborne pathogens varied depending on strains and specific amine. Cadaverine formation in control by L. monocytogenes and S. aureus were 500.9 and 948.1 mg/L, respectively while the CFSs of LAB induced 4-fold lower cadaverine production by L. monocytogenes and 7-fold lower cadaverine production by S. aureus. CFSs resulted in strong decreases in cadaverine and putrescine production by L. monocytogenes and S. aureus, although remarkable increases were observed for histamine, spermidine, spermine, serotonin, dopamine, tyramine, and agmatine, in the presence of LAB in lysine decarboxylase broth.Keywords: cell-free solution, lactic acid bacteria, cadaverine, food borne-pathogen
Procedia PDF Downloads 5404226 Cytotoxicological Evaluation of a Folate Receptor Targeting Drug Delivery System Based on Cyclodextrins
Authors: Caroline Mendes, Mary McNamara, Orla Howe
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For chemotherapy, a drug delivery system should be able to specifically target cancer cells and deliver the therapeutic dose without affecting normal cells. Folate receptors (FR) can be considered key targets since they are commonly over-expressed in cancer cells and they are the molecular marker used in this study. Here, cyclodextrin (CD) has being studied as a vehicle for delivering the chemotherapeutic drug, methotrexate (MTX). CDs have the ability to form inclusion complexes, in which molecules of suitable dimensions are included within the CD cavity. In this study, β-CD has been modified using folic acid so as to specifically target the FR molecular marker. Thus, the system studied here for drug delivery consists of β-CD, folic acid and MTX (CDEnFA:MTX). Cellular uptake of folic acid is mediated with high affinity by folate receptors while the cellular uptake of antifolates, such as MTX, is mediated with high affinity by the reduced folate carriers (RFCs). This study addresses the gene (mRNA) and protein expression levels of FRs and RFCs in the cancer cell lines CaCo-2, SKOV-3, HeLa, MCF-7, A549 and the normal cell line BEAS-2B, quantified by real-time polymerase chain reaction (real-time PCR) and flow cytometry, respectively. From that, four cell lines with different levels of FRs, were chosen for cytotoxicity assays of MTX and CDEnFA:MTX using the MTT assay. Real-time PCR and flow cytometry data demonstrated that all cell lines ubiquitously express moderate levels of RFC. These experiments have also shown that levels of FR protein in CaCo-2 cells are high, while levels in SKOV-3, HeLa and MCF-7 cells are moderate. A549 and BEAS-2B cells express low levels of FR protein. FRs are highly expressed in all the cancer cell lines analysed when compared to the normal cell line BEAS-2B. The cell lines CaCo-2, MCF-7, A549 and BEAS-2B were used in the cell viability assays. 48 hours treatment with the free drug and the complex resulted in IC50 values of 93.9 µM ± 9.2 and 56.0 µM ± 4.0 for CaCo-2 for free MTX and CDEnFA:MTX respectively, 118.2 µM ± 10.8 and 97.8 µM ± 12.3 for MCF-7, 36.4 µM ± 6.9 and 75.0 µM ± 8.5 for A549 and 132.6 µM ± 12.1 and 288.1 µM ± 16.3 for BEAS-2B. These results demonstrate that MTX is more toxic towards cell lines expressing low levels of FR, such as the BEAS-2B. More importantly, these results demonstrate that the inclusion complex CDEnFA:MTX showed greater cytotoxicity than the free drug towards the high FR expressing CaCo-2 cells, indicating that it has potential to target this receptor, enhancing the specificity and the efficiency of the drug.Keywords: cyclodextrins, cancer treatment, drug delivery, folate receptors, reduced folate carriers
Procedia PDF Downloads 3004225 iPSCs More Effectively Differentiate into Neurons on PLA Scaffolds with High Adhesive Properties for Primary Neuronal Cells
Authors: Azieva A. M., Yastremsky E. V., Kirillova D. A., Patsaev T. D., Sharikov R. V., Kamyshinsky R. A., Lukanina K. I., Sharikova N. A., Grigoriev T. E., Vasiliev A. L.
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Adhesive properties of scaffolds, which predominantly depend on the chemical and structural features of their surface, play the most important role in tissue engineering. The basic requirements for such scaffolds are biocompatibility, biodegradation, high cell adhesion, which promotes cell proliferation and differentiation. In many cases, synthetic polymers scaffolds have proven advantageous because they are easy to shape, they are tough, and they have high tensile properties. The regeneration of nerve tissue still remains a big challenge for medicine, and neural stem cells provide promising therapeutic potential for cell replacement therapy. However, experiments with stem cells have their limitations, such as low level of cell viability and poor control of cell differentiation. Whereas the study of already differentiated neuronal cell culture obtained from newborn mouse brain is limited only to cell adhesion. The growth and implantation of neuronal culture requires proper scaffolds. Moreover, the polymer scaffolds implants with neuronal cells could demand specific morphology. To date, it has been proposed to use numerous synthetic polymers for these purposes, including polystyrene, polylactic acid (PLA), polyglycolic acid, and polylactide-glycolic acid. Tissue regeneration experiments demonstrated good biocompatibility of PLA scaffolds, despite the hydrophobic nature of the compound. Problem with poor wettability of the PLA scaffold surface could be overcome in several ways: the surface can be pre-treated by poly-D-lysine or polyethyleneimine peptides; roughness and hydrophilicity of PLA surface could be increased by plasma treatment, or PLA could be combined with natural fibers, such as collagen or chitosan. This work presents a study of adhesion of both induced pluripotent stem cells (iPSCs) and mouse primary neuronal cell culture on the polylactide scaffolds of various types: oriented and non-oriented fibrous nonwoven materials and sponges – with and without the effect of plasma treatment and composites with collagen and chitosan. To evaluate the effect of different types of PLA scaffolds on the neuronal differentiation of iPSCs, we assess the expression of NeuN in differentiated cells through immunostaining. iPSCs more effectively differentiate into neurons on PLA scaffolds with high adhesive properties for primary neuronal cells.Keywords: PLA scaffold, neurons, neuronal differentiation, stem cells, polylactid
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