Search results for: recovery of cells
4339 Fabrication of Graphene Oxide Based Planar Hetero-Junction Perovskite Solar Cells
Authors: Khursheed Ahmad, Shaikh M. Mobin
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In this work, we have developed a highly stable planar heterojunction perovskite solar cells (PSCs) with a architecture (ITO/GO/PEDOT:PSS/MAPbI3/PCBM/Carbon tape). The PSCs was fabricated under air using GO/PEDOT:PSS as hole transport layer while the carbon tape used as a back contact to complete the device. The fabricated PSCs device exhibited good stability and performance in terms of power conversion efficiency of 5.2%. The PSCs devices were exposed to ambient condition for 4 days which shows excellent stability confirmed by XRD analysis. We believed that the stability of the planar heterojunction perovskite solar cell may be due the presence of GO which inhibits the direct contact between PEDOT:PSS and MAPbI3.Keywords: graphene oxide, perovskite solar cells, hole transport layer, PEDOT:PSS
Procedia PDF Downloads 1794338 Temporal Changes of Heterogeneous Subpopulations of Human Adipose-Derived Stromal/Stem Cells in vitro
Authors: Qiuyue Peng, Vladimir Zachar
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The application of adipose-derived stromal/stem cells (ASCs) in regenerative medicine is gaining more awareness due to their advanced translational potential and abundant source preparations. However, ASC-based translation has been confounded by high subpopulation heterogeneity, causing ambiguity about its precise therapeutic value. Some phenotypes defined by a unique combination of positive and negative surface markers have been found beneficial to the required roles. Therefore, the immunophenotypic repertoires of cultured ASCs and temporal changes of distinct subsets were investigated in this study. ASCs from three donors undergoing cosmetic liposuction were cultured in standard culturing methods, and the co-expression patterns based on the combination of selected markers at passages 1, 4, and 8 were analyzed by multi-chromatic flow cytometry. The results showed that the level of heterogeneity of subpopulations of ASCs became lower by in vitro expansion. After a few passages, most of the CD166⁺/CD274⁺/CD271⁺ based subpopulations converged to CD166 single positive cells. Meanwhile, these CD29⁺CD201⁺ double-positive cells, in combination with CD36/Stro-1 expression or without, feathered only the major epitopes and maintained prevailing throughout the whole process. This study suggested that, upon in vitro expansion, the phenotype repertoire of ASCs redistributed and stabilized in a way that cells co-expressing exclusively the strong markers remained dominant. These preliminary findings provide a general overview of the distribution of heterogeneous subsets residents within human ASCs during expansion in vitro. It is a critical step to fully characterize ASCs before clinical application, although the biological effects of heterogeneous subpopulations still need to be clarified.Keywords: adipose-derived stromal/stem cells, heterogeneity, immunophenotype, subpopulations
Procedia PDF Downloads 1084337 Rotor Concepts for the Counter Flow Heat Recovery Fan
Authors: Christoph Speer
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Decentralized ventilation systems should combine a small and economical design with high aerodynamic and thermal efficiency. The Counter Flow Heat Recovery Fan (CHRF) provides the ability to meet these requirements by using only one cross flow fan with a large number of blades to generate both airflows and which simultaneously acts as a regenerative counter flow heat exchanger. The successful development of the first laboratory prototype has shown the potential of this ventilation system. Occurring condensate on the surfaces of the fan blades during the cold and dry season can be recovered through the characteristic mode of operation. Hence the CHRF provides the possibility to avoid the need for frost protection and condensate drain. Through the implementation of system-specific solutions for flow balancing and summer bypass the required functionality is assured. The scalability of the CHRF concept allows the use in renovation as well as in new buildings from single-room devices through to systems for office buildings. High aerodynamic and thermal efficiency and the lower number of required mechatronic components should enable a reduction in investment as well as operating costs. The rotor is the key component of the system, the requirements and possible implementation variants are presented.Keywords: CHRF, counter flow heat recovery fan, decentralized ventilation system, renovation
Procedia PDF Downloads 3514336 Peptidoglycan Vaccine-On-Chip against a Lipopolysaccharide-Induced Experimental Sepsis Model
Authors: Katerina Bakela, Ioanna Zerva, Irene Athanassakis
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Lipopolysaccharide (LPS) is commonly used in murine sepsis models, which are largely associated with immunosuppression (incretion of MDSCs cells and Tregs, imbalance of inflammatory/anti-inflammatory cytokines) and collapse of the immune system. After adapting the LPS treatment to the needs of locally bred BALB/c mice, the present study explored the protective role of Micrococcus luteus peptidoglycan (PG) pre-activated vaccine-on chip in endotoxemia. The established protocol consisted of five daily intraperitoneal injections of 0.2mg/g LPS. Such protocol allowed longer survival, necessary in the prospect of the therapeutic treatment application. The so-called vaccine-on-chip consists of a 3-dimensional laser micro-texture Si-scaffold loaded with BALB/c mouse macrophages and activated in vitro with 1μg/ml PG, which exert its action upon subcutaneous implantation. The LPS treatment significantly decreased CD4+, CD8+, CD3z+, and CD19+ cells, while increasing myeloid-derived suppressor cells (MDSCs), CD25+, and Foxp3+ cells. These results were accompanied by increased arginase-1 activity in spleen cell lysates and production of IL-6, TNF-a, and IL-18 while acquiring severe sepsis phenotype as defined by the murine sepsis scoring. The in vivo application of PG pre-activated vaccine-on chip significantly decreased the percent of CD11b+, Gr1+, CD25+, Foxp3+ cells, and arginase-1 activity in the spleen of LPS-treated animals, while decreasing IL-6 and TNF-a in the serum, allowing survival to all animals tested and rescuing the severity of sepsis phenotype. In conclusion, these results reveal a promising mode of action of PG pre-activated vaccine-on chip in LPS endotoxemia, strengthening; thus, the use of treatment is septic patients.Keywords: myeloid-derived suppressor cells, peptidoglycan, sepsis, Si-scaffolds
Procedia PDF Downloads 1344335 Comparison of Bismuth-Based Nanoparticles as Radiosensitization Agents for Radiotherapy
Authors: Merfat Algethami, Anton Blencowe, Bryce Feltis, Stephen Best, Moshi Geso
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Nano-materials with high atomic number atoms have been demonstrated to enhance the effective radiation dose and thus potentially could improve therapeutic efficacy in radiotherapy. The optimal nanoparticulate agents require high X-ray absorption coefficients, low toxicity, and should be cost effective. The focus of our research is the development of a nanoparticle therapeutic agent that can be used in radiotherapy to provide optimal enhancement of the radiation effects on the target. In this study, we used bismuth (Bi) nanoparticles coated with starch and bismuth sulphide nanoparticles (Bi2S3) coated with polyvinylpyrrolidone (PVP). These NPs are of low toxicity and are one of the least expensive heavy metal-based nanoparticles. The aims of this study were to synthesise Bi2S3 and Bi NPs, and examine their cytotoxicity to human lung adenocarcinoma epithelial cells (A549). The dose enhancing effects of NPs on A549 cells were examined at both KV and MV energies. The preliminary results revealed that bismuth based nanoparticles show increased radio-sensitisation of cells, displaying dose enhancement with KV X-ray energies and to a lesser degree for the MV energies. We also observed that Bi NPs generated a greater dose enhancement effect than Bi2S3 NPs in irradiated A549 cells. The maximum Dose Enhancement Factor (DEF) was obtained at lower energy KV range when cells treated with Bi NPs (1.5) compared to the DEF of 1.2 when cells treated with Bi2S3NPs. Less radiation dose enhancement was observed when using high energy MV beam with higher DEF value of Bi NPs treatment (1.26) as compared to 1.06 DEF value with Bi2S3 NPs. The greater dose enhancement was achieved at KV energy range, due the effect of the photoelectric effect which is the dominant process of interaction of X-ray. The cytotoxic effect of Bi NPs on enhancing the X-ray dose was higher due to the higher amount of elemental Bismuth present in Bi NPs compared to Bi2S3 NPs. The results suggest that Bismuth based NPs can be considered as valuable dose enhancing agents when used in clinical applications.Keywords: A549 lung cancer cells, Bi2S3 nanoparticles, dose enhancement effect, radio-sensitising agents
Procedia PDF Downloads 2704334 Potential Activities of Human Endogenous Retroviral kDNA in Melanoma Pathogenesis and HIV-1 Infection
Authors: Jianli Dong, Fangling Xu, Gengming Huang
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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
Procedia PDF Downloads 2314333 Effect of the Polymer Modification on the Cytocompatibility of Human and Rat Cells
Authors: N. Slepickova Kasalkova, P. Slepicka, L. Bacakova, V. Svorcik
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Tissue engineering includes combination of materials and techniques used for the improvement, repair or replacement of the tissue. Scaffolds, permanent or temporally material, are used as support for the creation of the "new cell structures". For this important component (scaffold), a variety of materials can be used. The advantage of some polymeric materials is their cytocompatibility and possibility of biodegradation. Poly(L-lactic acid) (PLLA) is a biodegradable, semi-crystalline thermoplastic polymer. PLLA can be fully degraded into H2O and CO2. In this experiment, the effect of the surface modification of biodegradable polymer (performed by plasma treatment) on the various cell types was studied. The surface parameters and changes of the physicochemical properties of modified PLLA substrates were studied by different methods. Surface wettability was determined by goniometry, surface morphology and roughness study were performed with atomic force microscopy and chemical composition was determined using photoelectron spectroscopy. The physicochemical properties were studied in relation to cytocompatibility of human osteoblast (MG 63 cells), rat vascular smooth muscle cells (VSMC), and human stem cells (ASC) of the adipose tissue in vitro. A fluorescence microscopy was chosen to study and compare cell-material interaction. Important parameters of the cytocompatibility like adhesion, proliferation, viability, shape, spreading of the cells were evaluated. It was found that the modification leads to the change of the surface wettability depending on the time of modification. Short time of exposition (10-120 s) can reduce the wettability of the aged samples, exposition longer than 150 s causes to increase of contact angle of the aged PLLA. The surface morphology is significantly influenced by duration of modification, too. The plasma treatment involves the formation of the crystallites, whose number increases with increasing time of modification. On the basis of physicochemical properties evaluation, the cells were cultivated on the selected samples. Cell-material interactions are strongly affected by material chemical structure and surface morphology. It was proved that the plasma treatment of PLLA has a positive effect on the adhesion, spreading, homogeneity of distribution and viability of all cultivated cells. This effect was even more apparent for the VSMCs and ASCs which homogeneously covered almost the whole surface of the substrate after 7 days of cultivation. The viability of these cells was high (more than 98% for VSMCs, 89-96% for ASCs). This experiment is one part of the basic research, which aims to easily create scaffolds for tissue engineering with subsequent use of stem cells and their subsequent "reorientation" towards the bone cells or smooth muscle cells.Keywords: poly(L-lactic acid), plasma treatment, surface characterization, cytocompatibility, human osteoblast, rat vascular smooth muscle cells, human stem cells
Procedia PDF Downloads 2274332 Cognitive Model of Analogy Based on Operation of the Brain Cells: Glial, Axons and Neurons
Authors: Ozgu Hafizoglu
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Analogy is an essential tool of human cognition that enables connecting diffuse and diverse systems with attributional, deep structural, casual relations that are essential to learning, to innovation in artificial worlds, and to discovery in science. Cognitive Model of Analogy (CMA) leads and creates information pattern transfer within and between domains and disciplines in science. This paper demonstrates the Cognitive Model of Analogy (CMA) as an evolutionary approach to scientific research. The model puts forward the challenges of deep uncertainty about the future, emphasizing the need for flexibility of the system in order to enable reasoning methodology to adapt to changing conditions. In this paper, the model of analogical reasoning is created based on brain cells, their fractal, and operational forms within the system itself. Visualization techniques are used to show correspondences. Distinct phases of the problem-solving processes are divided thusly: encoding, mapping, inference, and response. The system is revealed relevant to brain activation considering each of these phases with an emphasis on achieving a better visualization of the brain cells: glial cells, axons, axon terminals, and neurons, relative to matching conditions of analogical reasoning and relational information. It’s found that encoding, mapping, inference, and response processes in four-term analogical reasoning are corresponding with the fractal and operational forms of brain cells: glial, axons, and neurons.Keywords: analogy, analogical reasoning, cognitive model, brain and glials
Procedia PDF Downloads 1844331 Effect of Wettability Alteration in Low Salt Water Injection Modeling
Authors: H. Vahdani
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By the adsorption of polar compounds and/or the deposition of organic material, the wettability of originally water-wet reservoir rock can be altered. The degree of alteration is determined by the interaction of the oil constituents, the mineral surface, and the brine chemistry. Recently improving oil recovery by tuning wettability alteration is believed as a new recovery method. Various researchers have demonstrated that low salt water injection has a significant impact on oil recovery. It has been shown, for instance, that additional oil can be produced from reservoir rock by managing the injection water. Large wettability sensitivity has been observed, indicating that the oil/water capillary pressure profiles play a major role during low saline water injection simulation. Although the exact physics on how this alteration occurs is still a research topic; however, it has been reported that some of its effect can be captured by a relative permeability shift from an oil-wet system to a water-wet system. Modeling of low salt water injection mainly is based on the theory of wettability alteration and is hence strongly dependent on the wettability of the reservoir. In this article, combination of different wettabilities has been simulated and it is observed that the highest recoveries were from the cases were the reservoir initially was water-wet, and the lowest recoveries was from the cases were the reservoir initially was considered oil-wet. However for the cases where the reservoir initially was oil-wet, the effect of low-salinity waterflooding was the largest.Keywords: low salt water injection, wettability alteration, modelling, relative permeability
Procedia PDF Downloads 4944330 CO₂/CH₄ Exchange Studies on Shales to Assess the Potential for CO₂ Storage and Enhanced Shale Gas Recovery
Authors: Mateusz Kudasik, Katarzyna Kozieł
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The work included detailed studies of CO₂/CH₄ exchange on a shale core from the Lewino-1G2 well (Poland) from a depth of 3408 m. The sample permeability coefficients were determined under conditions of confining pressure from 5 MPa to 35 MPa. These studies showed that at a confining pressure of 35 MPa – corresponding to a depth of about 1000 m, the shale core was impermeable in the direction perpendicular to the bedding, and in the direction parallel to the bedding, the sample had very low permeability (k∞=0.001 mD). The sorption tests performed showed low sorption capacities, which amounted to a maximum of 1.28 cm³/g in relation to CO₂ and 0.87 cm³/g to CH₄ at a pressure of 1.4 MPa. The most important study used to assess the possibilities of CO₂ storage and gas recovery from shale rocks were the CO₂/CH₄ exchange experiments, which were carried out under confining pressure conditions of 5 MPa and 30 MPa. These experiments were carried out on a unique apparatus, which makes it possible to apply a confining pressure corresponding to in situ conditions. The obtained results made it possible to carry out a comprehensive balance of gas exchange during the injection of CO₂ into the shale sample, with simultaneous recovery of CH₄. Based on the conducted sorption and gas exchange studies on the core sample under confining pressure conditions, it was found that in situ conditions, at the depths of shale gas occurrence in Poland of 3000-4000 m, where the confining pressure can be about 100 MPa: (i) poorly developed pore structure, (ii) very low permeability, and (iii) low sorption properties, make shale rocks poorly predisposed to the application of CO₂ storage technology with simultaneous recovery of CH₄. Without the stimulation of CO₂/CH₄ exchange rates through fracturing processes, the effectiveness of CO₂-ESGR technology on shale rock is very low. The research presented in this work is extremely important from the point of view of precise assessment of the potential of CO₂-ESGR technology.Keywords: shale gas, shale rocks, CO₂/CH₄ exchange, permeability, sorption, CO₂, CH₄
Procedia PDF Downloads 44329 Review of Sulfur Unit Capacity Expansion Options
Authors: Avinashkumar Karre
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Sulfur recovery unit, most commonly called as Claus process, is very significant gas desulfurization process unit in refinery and gas industries. Explorations of new natural gas fields, refining of high-sulfur crude oils, and recent crude expansion projects are needing capacity expansion of Claus unit for many companies around the world. In refineries, the sulphur recovery units take acid gas from amine regeneration units and sour water strippers, converting hydrogen sulfide to elemental sulfur using the Claus process. The Claus process is hydraulically limited by mass flow rate. Reducing the pressure drop across control valves, flow meters, lines, knock-out drums, and packing improves the capacity. Oxygen enrichment helps improve the capacity by removing nitrogen, this is more commonly done on all capacity expansion projects. Typical upgrades required due to oxygen enrichment are new burners, new refractory in thermal reactor, resizing of 1st condenser, instrumentation changes, and steam/condensate heat integration. Some other capacity expansion options typically considered are tail gas compressor, replacing air blower with higher head, hydrocarbon minimization in the feed, water removal, and ammonia removal. Increased capacity related upgrades in sulfur recovery unit also need changes in the tail gas treatment unit, typical changes include improvement to quench tower duty, packing area upgrades in quench and absorber towers and increased amine circulation flow rates.Keywords: Claus process, oxygen enrichment, sulfur recovery unit, tail gas treatment unit
Procedia PDF Downloads 1234328 Study of Mechanical Properties of Large Scale Flexible Silicon Solar Modules on the Various Substrates
Authors: M. Maleczek, Leszek Bogdan, Kazimierz Drabczyk, Agnieszka Iwan
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Crystalline silicon (Si) solar cells are the main product in the market among the various photovoltaic technologies concerning such advantages as: material richness, high carrier mobilities, broad spectral absorption range and established technology. However, photovoltaic technology on the stiff substrates are heavier, more fragile and less cost-effective than devices on the flexible substrates to be applied in special applications. The main goal of our work was to incorporate silicon solar cells into various fabric, without any change of the electrical and mechanical parameters of devices. This work is realized for the GEKON project (No. GEKON2/O4/268473/23/2016) sponsored by The National Centre for Research and Development and The National Fund for Environmental Protection and Water Management. In our work, the polyamide or polyester fabrics were used as a flexible substrate in the created devices. Applied fabrics differ in tensile and tear strength. All investigated polyamide fabrics are resistant to weathering and UV, while polyester ones is resistant to ozone, water and ageing. The examined fabrics are tight at 100 cm water per 2 hours. In our work, commercial silicon solar cells with the size 156 × 156 mm were cut into nine parts (called single solar cells) by diamond saw and laser. Gap and edge after cutting of solar cells were checked by transmission electron microscope (TEM) to study morphology and quality of the prepared single solar cells. Modules with the size of 160 × 70 cm (containing about 80 single solar cells) were created and investigated by electrical and mechanical methods. Weight of constructed module is about 1.9 kg. Three types of solar cell architectures such as: -fabric/EVA/Si solar cell/EVA/film for lamination, -backsheet PET/EVA/Si solar cell/EVA/film for lamination, -fabric/EVA/Si solar cell/EVA/tempered glass, were investigated taking into consideration type of fabric and lamination process together with the size of solar cells. In investigated devices EVA, it is ethylene-vinyl acetate, while PET - polyethylene terephthalate. Depend on the lamination process and compatibility of textile with solar cell an efficiency of investigated flexible silicon solar cells was in the range of 9.44-16.64 %. Multi folding and unfolding of flexible module has no impact on its efficiency as was detected by Instron equipment. Power (P) of constructed solar module is 30 W, while voltage about 36 V. Finally, solar panel contains five modules with the polyamide fabric and tempered glass will be produced commercially for different applications (dual use).Keywords: flexible devices, mechanical properties, silicon solar cells, textiles
Procedia PDF Downloads 1724327 Research on the Role of Platelet Derived Growth Factor Receptor Beta in Promoting Dedifferentiation and Pulmonary Metastasis of Osteosarcoma Under Hypoxic Microenvironment
Authors: Enjie Xu, Zhen Huang, Kunpeng Zhu, Jianping Hu, Xiaolong Ma, Yongjie Wang, Jiazhuang Zhu, Chunlin Zhang
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Abstract: Hypoxia and dedifferentiation of osteosarcoma (OS) cells leads to poor prognosis. We plan to identify the role of hypoxia on dedifferentiation and the associated signaling pathways. We performed a sphere formation assay and determined spheroid cells as dedifferentiated cells by detecting stem cell-like markers. RNAi assay was used to explore the expression relationship between hypoxia inducible factor 1 subunit alpha (HIF1A) and platelet derived growth factor receptor beta (PDGFRB). We obtained PDGFRB knockdown and overexpression cells through lentiviral infection experiments and the effects of PDGFRB on cytoskeleton rearrangement and cell adhesion were explored by immunocytochemistry. Wound-healing experiments, transwell assays, and animal trials were employed to investigate the effect of PDGFRB on OS metastasis. Dedifferentiated OS cells were found to exhibit high expression of HIF1A and PDGFRB, and HIF1A promoted the expression of PDGFRB, subsequently activated ras homolog family member A (RhoA), and increased the phosphorylation of myosin light chain (MLC). PDGFRB also enhanced the phosphorylation of focal adhesion kinase (FAK). The OS cell morphology and vinculin distribution were altered by PDGFRB. PDGFRB also promoted cell dedifferentiation and had a significant impact on the metastasis of OS cells both in vitro and in vivo. Our results demonstrated that HIF1A up-regulated PDGFRB under hypoxic conditions, and PDGFRB regulated the actin cytoskeleton by activating RhoA and subsequently phosphorylating MLC, thereby promoting OS dedifferentiation and pulmonary metastasis.Keywords: osteosarcoma, dedifferentiation, metastasis, cytoskeleton rearrangement, PDGFRB, hypoxia
Procedia PDF Downloads 464326 Research on Low interfacial Tension Viscoelastic Fluid Oil Displacement System in Unconventional Reservoir
Authors: Long Long Chen, Xinwei Liao, Shanfa Tang, Shaojing Jiang, Ruijia Tang, Rui Wang, Shu Yun Feng, Si Yao Wang
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Unconventional oil reservoirs have the characteristics of strong heterogeneity and poor injectability, and traditional chemical flooding technology is not effective in such reservoirs; polymer flooding in the production of heavy oil reservoirs is difficult to handle produced fluid and easy to block oil wells, etc. Therefore, a viscoelastic fluid flooding system with good adaptability, low interfacial tension, plugging, and diverting capabilities was studied. The viscosity, viscoelasticity, surface/interfacial activity, wettability, emulsification, and oil displacement performance of the anionic Gemini surfactant flooding system were studied, and the adaptability of the system to the reservoir environment was evaluated. The oil displacement effect of the system in low-permeability and high-permeability (heavy oil) reservoirs was investigated, and the mechanism of the system to enhance water flooding recovery was discussed. The results show that the system has temperature resistance and viscosity increasing performance (65℃, 4.12mPa•s), shear resistance and viscoelasticity; at a lower concentration (0.5%), the oil-water interfacial tension can be reduced to ultra-low (10-3mN/m); has good emulsifying ability for heavy oil, and is easy to break demulsification (4.5min); has good adaptability to reservoirs with high salinity (30000mg/L). Oil flooding experiments show that this system can increase the water flooding recovery rate of low-permeability homogeneous and heterogeneous cores by 13% and 15%, respectively, and can increase the water-flooding recovery rate of high-permeability heavy oil reservoirs by 40%. The anionic Gemini surfactant flooding system studied in this paper is a viscoelastic fluid, has good emulsifying and oil washing ability, can effectively improve sweep efficiency, reduce injection pressure, and has broad application in unconventional reservoirs to enhance oil recovery prospect.Keywords: oil displacement system, recovery factor, rheology, interfacial activity, environmental adaptability
Procedia PDF Downloads 1224325 Efficacy of Light-Emitting Diode-Mediated Photobiomodulation in Tendon Healing in a Murine Model
Authors: Sukwoong Kang
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Background: The application of light-emitting diode (LED)-dependent photobiomodulation (PBM) in promoting post-tendon injury healing has been recently reported. Despite the establishment of a theoretical basis for ligament restoration through PBM, the lack of any empirical evidence deems this therapeutic strategy contentious. Therefore, the aim of this study was to investigate the potency of LED-based PBM in facilitating tendon healing in a murine model. Methods: Migration kinetics were analyzed at two specific wavelengths: 630 and 880 nm. The Achilles tendon in the hind limbs of Balb/c mice was severed via Achilles tendon transection. Subsequently, the mice were randomized into LED non-irradiation and LED irradiation groups. Mice with intact tendons were employed as healthy controls. The wounds were LED-irradiated for 20 min daily for two days. Histological properties, tendon healing mediators, and inflammatory mediators were screened on day 14. Results: The roundness of the nuclei and fiber structure, indicating the degree of infiltrated inflammatory cells and severity of fiber fragmentation, respectively, were considerably lower in the LED irradiation group than in the LED non-irradiation group. Immunohistochemical analysis depicted an increase in tenocytes (SCX+ cells) and a recovery of wounds with reduced fibrosis (lower collagen 3 and TGF-β1) in the LED irradiation group during healing; conversely, the LED non-irradiation group exhibited tissue fibrosis. The ratio of M2 macrophages to total macrophages was higher in the LED irradiation group than in the injured group. Conclusion: LED-based PBM in the Achilles tendon rupture murine model effectuated a rapid restoration of histological and immunochemical outcomes. The aforementioned findings suggest that LED-based PBM presents remarkable potential as an adjunct therapeutic for tendon healing and warrants further research to standardize various parameters to advance and establish it as a reliable treatment regime.Keywords: photobiomodulation, light-emitting diode, tendon, regeneration
Procedia PDF Downloads 434324 Study of the Stability of Underground Mines by Numerical Method: The Mine Chaabet El Hamra, Algeria
Authors: Nakache Radouane, M. Boukelloul, M. Fredj
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Method room and pillar sizes are key factors for safe mining and their recovery in open-stop mining. This method is advantageous due to its simplicity and requirement of little information to be used. It is probably the most representative method among the total load approach methods although it also remains a safe design method. Using a finite element software (PLAXIS 3D), analyses were carried out with an elasto-plastic model and comparisons were made with methods based on the total load approach. The results were presented as the optimization for improving the ore recovery rate while maintaining a safe working environment.Keywords: room and pillar, mining, total load approach, elasto-plastic
Procedia PDF Downloads 3284323 Stem Cell Fate Decision Depending on TiO2 Nanotubular Geometry
Authors: Jung Park, Anca Mazare, Klaus Von Der Mark, Patrik Schmuki
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In clinical application of TiO2 implants on tooth and hip replacement, migration, adhesion and differentiation of neighboring mesenchymal stem cells onto implant surfaces are critical steps for successful bone regeneration. In a recent decade, accumulated attention has been paid on nanoscale electrochemical surface modifications on TiO2 layer for improving bone-TiO2 surface integration. We generated, on titanium surfaces, self-assembled layers of vertically oriented TiO2 nanotubes with defined diameters between 15 and 100 nm and here we show that mesenchymal stem cells finely sense TiO2 nanotubular geometry and quickly decide their cell fate either to differentiation into osteoblasts or to programmed cell death (apoptosis) on TiO2 nanotube layers. These cell fate decisions are critically dependent on nanotube size differences (15-100nm in diameters) of TiO2 nanotubes sensing by integrin clustering. We further demonstrate that nanoscale topography-sensing is feasible not only in mesenchymal stem cells but rather seems as generalized nanoscale microenvironment-cell interaction mechanism in several cell types composing bone tissue network including osteoblasts, osteoclast, endothelial cells and hematopoietic stem cells. Additionally we discuss the synergistic effect of simultaneous stimulation by nanotube-bound growth factor and nanoscale topographic cues on enhanced bone regeneration.Keywords: TiO2 nanotube, stem cell fate decision, nano-scale microenvironment, bone regeneration
Procedia PDF Downloads 4274322 The Nature of the Complicated Fabric Textures: How to Represent in Primary Visual Cortex
Authors: J. L. Liu, L. Wang, B. Zhu, J. Zhou, W. D. Gao
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Fabric textures are very common in our daily life. However, we never explore the representation of fabric textures from neuroscience view. Theoretical studies suggest that primary visual cortex (V1) uses a sparse code to efficiently represent natural images. However, how the simple cells in V1 encode the artificial textures is still a mystery. So, here we will take fabric texture as stimulus to study the response of independent component analysis that is established to model the receptive field of simple cells in V1. Experimental results based on 140 classical fabric images indicate that the receptive fields of simple cells have obvious selectivity in orientation, frequency, and phase when drifting gratings are used to determine their tuning properties. Additionally, the distribution of optimal orientation and frequency shows that the patch size selected from each original fabric image has a significant effect on the frequency selectivity.Keywords: fabric texture, receptive filed, simple cell, spare coding
Procedia PDF Downloads 4744321 Investigation and Estimation of State of Health of Battery Pack in Battery Electric Vehicles-Online Battery Characterization
Authors: Ali Mashayekh, Mahdiye Khorasani, Thomas Weyh
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The tendency to use the Battery-Electric vehicle (BEV) for the low and medium driving range or even high driving range has been growing more and more. As a result, higher safety, reliability, and durability of the battery pack as a component of electric vehicles, which has a great share of cost and weight of the final product, are the topics to be considered and investigated. Battery aging can be considered as the predominant factor regarding the reliability and durability of BEV. To better understand the aging process, offline battery characterization has been widely used, which is time-consuming and needs very expensive infrastructures. This paper presents the substitute method for the conventional battery characterization methods, which is based on battery Modular Multilevel Management (BM3). According to this Topology, the battery cells can be drained and charged concerning their capacity, which allows varying battery pack structures. Due to the integration of the power electronics, the output voltage of the battery pack is no longer fixed but can be dynamically adjusted in small steps. In other words, each cell can have three different states, namely series, parallel, and bypass in connection with the neighbor cells. With the help of MATLAB/Simulink and by using the BM3 modules, the battery string model is created. This model allows us to switch two cells with the different SoC as parallel, which results in the internal balancing of the cells. But if the parallel switching lasts just for a couple of ms, we can have a perturbation pulse which can stimulate the cells out of the relaxation phase. With the help of modeling the voltage response pulse of the battery, it would be possible to characterize the cell. The Online EIS method, which is discussed in this paper, can be a robust substitute for the conventional battery characterization methods.Keywords: battery characterization, SoH estimation, RLS, BEV
Procedia PDF Downloads 1474320 Histone Deacetylases Inhibitor - Valproic Acid Sensitizes Human Melanoma Cells for alkylating agent and PARP inhibitor
Authors: Małgorzata Drzewiecka, Tomasz Śliwiński, Maciej Radek
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The inhibition of histone deacetyles (HDACs) holds promise as a potential anti-cancer therapy because histone and non-histone protein acetylation is frequently disrupted in cancer, leading to cancer initiation and progression. Additionally, histone deacetylase inhibitors (HDACi) such as class I HDAC inhibitor - valproic acid (VPA) have been shown to enhance the effectiveness of DNA-damaging factors, such as cisplatin or radiation. In this study, we found that, using of VPA in combination with talazoparib (BMN-637 – PARP1 inhibitor – PARPi) and/or Dacarabazine (DTIC - alkylating agent) resulted in increased DNA double strand break (DSB) and reduced survival (while not affecting primary melanocytes )and proliferation of melanoma cells. Furthermore, pharmacologic inhibition of class I HDACs sensitizes melanoma cells to apoptosis following exposure to DTIC and BMN-637. In addition, inhibition of HDAC caused sensitization of melanoma cells to dacarbazine and BMN-637 in melanoma xenografts in vivo. At the mRNA and protein level histone deacetylase inhibitor downregulated RAD51 and FANCD2. This study provides that combining HDACi, alkylating agent and PARPi could potentially enhance the treatment of melanoma, which is known for being one of the most aggressive malignant tumors. The findings presented here point to a scenario in which HDAC via enhancing the HR-dependent repair of DSBs created during the processing of DNA lesions, are essential nodes in the resistance of malignant melanoma cells to methylating agent-based therapies.Keywords: melanoma, hdac, parp inhibitor, valproic acid
Procedia PDF Downloads 814319 PDMS-Free Microfluidic Chips Fabrication and Utilisation for Pulsed Electric Fields Applications
Authors: Arunas Stirke, Neringa Bakute, Gatis Mozolevskis
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A technology of microfluidics is an emerging tool in the field of biology, medicine and chemistry. Microfluidic device is also known as ‘lab-on-a-chip’ technology [1]. In moving from macro- to microscale, there is unprecedented control over spatial and temporal gradients and patterns that cannot be captured in conventional Petri dishes and well plates [2]. However, there is not a single standard microfluidic chip designated for all purposes – every different field of studies needs a specific microchip with certain geometries, inlet/outlet, channel depth and other parameters to precisely regulate the required function. Since our group is studying an effect of pulsed electric field (PEF) to the cells, we have manufactured a microfluidic chip designated for high-throughput electroporation of cells. In our microchip, a cell culture chamber is divided into two parallel channels by a membrane, meanwhile electrodes for electroporation are attached to the wall of the channels. Both microchannels have their own inlet and outlet, enabling injection of transfection material separately. Our perspective is to perform electroporation of mammalian cells in two different ways: (1) plasmid and cells are injected in the same microchannel and (2) injected into separate microchannels. Moreover, oxygen and pH sensors are integrated on order to analyse cell viability parameters after PEF treatment.Keywords: microfluidics, chip, fabrication, electroporation
Procedia PDF Downloads 824318 Culture of Human Mesenchymal Stem Cells Culture in Xeno-Free Serum-Free Culture Conditions on Laminin-521
Authors: Halima Albalushi, Mohadese Boroojerdi, Murtadha Alkhabori
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Introduction: Maintenance of stem cell properties during culture necessitates the recreation of the natural cell niche. Studies reported the promising outcome of mesenchymal stem cells (MSC) properties maintenance after using extracellular matrix such as CELLstart™, which is the recommended coating material for stem cells cultured in serum-free and xeno-free conditions. Laminin-521 is known as a crucial adhesion protein, which is found in natural stem cell niche, and plays an important role in facilitating the maintenance of self-renewal, pluripotency, standard morphology, and karyotype of human pluripotent stem cells (PSCs). The aim of this study is to investigate the effects of Laminin-521 on human umbilical cord-derived mesenchymal stem cells (UC-MSC) characteristics as a step toward clinical application. Methods: Human MSC were isolated from the umbilical cord via the explant method. Umbilical cord-derived-MSC were cultured in serum-free and xeno-free conditions in the presence of Laminin-521 for six passages. Cultured cells were evaluated by morphology and expansion index for each passage. Phenotypic characterization of UC-MSCs cultured on Laminin-521 was evaluated by assessment of cell surface markers. Results: Umbilical cord derived-MSCs formed small colonies and expanded as a homogeneous monolayer when cultured on Laminin-521. Umbilical cord derived-MSCs reached confluence after 4 days in culture. No statistically significant difference was detected in all passages when comparing the expansion index of UC-MSCs cultured on LN-521 and CELLstart™. Phenotypic characterization of UC-MSCs cultured on LN-521 using flow cytometry revealed positive expression of CD73, CD90, CD105 and negative expression of CD34, CD45, CD19, CD14 and HLA-DR.Conclusion: Laminin-521 is comparable to CELLstart™ in supporting UC-MSCs expansion and maintaining their characteristics during culture in xeno-free and serum-free culture conditions.Keywords: mesenchymal stem cells, culture, laminin-521, xeno-free serum-free
Procedia PDF Downloads 724317 Role of Endonuclease G in Exogenous DNA Stability in HeLa Cells
Authors: Vanja Misic, Mohamed El-Mogy, Yousef Haj-Ahmad
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Endonuclease G (EndoG) is a well conserved mitochondrio-nuclear nuclease with dual lethal and vital roles in the cell. The aim of our study was to examine whether EndoG exerts its nuclease activity on exogenous DNA substrates such as plasmid DNA (pDNA), considering their importance in gene therapy applications. The effects of EndoG knockdown on pDNA stability and levels of encoded reporter gene expression were evaluated in the cervical carcinoma HeLa cells. Transfection of pDNA vectors encoding short-hairpin RNAs (shRNAs) reduced levels of EndoG mRNA and nuclease activity in HeLa cells. In physiological circumstances, EndoG knockdown did not have an effect on the stability of pDNA or the levels of encoded transgene expression as measured over a four day time-course. However, when endogenous expression of EndoG was induced by an extrinsic stimulus, targeting of EndoG by shRNA improved the perceived stability and transgene expression of pDNA vectors. Therefore, EndoG is not a mediator of exogenous DNA clearance, but in non-physiological circumstances it may non-specifically cleave intracellular DNA regardless of its origin. These findings make it unlikely that targeting of EndoG is a viable strategy for improving the duration and level of transgene expression from non-viral DNA vectors in gene therapy efforts.Keywords: EndoG, silencing, exogenous DNA stability, HeLa cells
Procedia PDF Downloads 4594316 A Strategy to Oil Production Placement Zones Based on Maximum Closeness
Authors: Waldir Roque, Gustavo Oliveira, Moises Santos, Tatiana Simoes
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Increasing the oil recovery factor of an oil reservoir has been a concern of the oil industry. Usually, the production placement zones are defined after some analysis of geological and petrophysical parameters, being the rock porosity, permeability and oil saturation of fundamental importance. In this context, the determination of hydraulic flow units (HFUs) renders an important step in the process of reservoir characterization since it may provide specific regions in the reservoir with similar petrophysical and fluid flow properties and, in particular, techniques supporting the placement of production zones that favour the tracing of directional wells. A HFU is defined as a representative volume of a total reservoir rock in which petrophysical and fluid flow properties are internally consistent and predictably distinct of other reservoir rocks. Technically, a HFU is characterized as a rock region that exhibit flow zone indicator (FZI) points lying on a straight line of the unit slope. The goal of this paper is to provide a trustful indication for oil production placement zones for the best-fit HFUs. The FZI cloud of points can be obtained from the reservoir quality index (RQI), a function of effective porosity and permeability. Considering log and core data the HFUs are identified and using the discrete rock type (DRT) classification, a set of connected cell clusters can be found and by means a graph centrality metric, the maximum closeness (MaxC) cell is obtained for each cluster. Considering the MaxC cells as production zones, an extensive analysis, based on several oil recovery factor and oil cumulative production simulations were done for the SPE Model 2 and the UNISIM-I-D synthetic fields, where the later was build up from public data available from the actual Namorado Field, Campos Basin, in Brazil. The results have shown that the MaxC is actually technically feasible and very reliable as high performance production placement zones.Keywords: hydraulic flow unit, maximum closeness centrality, oil production simulation, production placement zone
Procedia PDF Downloads 3284315 Bioremediation of Arsenic from Industrially Polluted Soil of Vatva, Ahmedabad, Gujarat, India
Authors: C. Makwana, S. R. Dave
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Arsenic is toxic to almost all living cells. Its contamination in natural sources affects the growth of microorganisms. The presence of arsenic is associated with various human disorders also. The attempt of this sort of study provides information regarding the performance of our isolated microorganisms in the presence of Arsenic, which have ample scope for bioremediation. Six isolates were selected from the polluted sample of industrial zone Vatva, Ahmedabad, Gujarat, India, out of which two were Thermophilic organisms. The thermophilic exopolysaccharide (EPS) producing Bacillus was used for microbial enhance oil recovery (MEOR) and in the bio beneficiation. Inorganic arsenic primarily exists in the form of arsenate or arsenite. This arsenic resistance isolate was capable of transforming As +3 to As+5. This isolate would be useful for arsenic remediation standpoint from aquatic systems. The study revealed that the thermophilic microorganism was growing at 55 degree centigrade showed considerable remediation property. The results on the growth and enzyme catalysis would be discussed in response to Arsenic remediation.Keywords: aquatic systems, thermophilic, exopolysacchride, arsenic
Procedia PDF Downloads 2124314 Flotation of Rare Earth Oxides from Iron-Oxide Silicate Rich Tailings Using Fatty Acids
Authors: George B. Abaka-Wood, Massimiliano Zanin, Jonas Addai-Mensah, William Skinner
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The versatility of froth flotation has made it vital in the beneficiation of rare earth elements minerals from either high or low-grade ores. There has been a significant increase in the quantity of iron oxide silicate-rich tailings generated from the extraction of primary commodities such as copper and gold in Australia, which have been identified to contain very low-grade rare earth oxides (≤ 1%). There is a vast knowledge gap in the beneficiation of rare earth oxides from such tailings. The aim of this research is to investigate the feasibility of using fatty acids as collectors for the flotation recovery and upgrade of rare earth oxides from selected iron-oxide silicate-rich tailings. Two forms of fatty acid collectors (oleic acid and sodium oleate) were tested in this investigation. Flotation tests were carried out using a 1.2 L Denver D-12 cell. The effects of pulp pH, fatty acid dosage, particle size distribution (-150 +75 µm, -75 +38 µm and -38 µm) and conventional depressants (sodium silicate and starch) dosage on flotation recovery of rare earth oxides were investigated. A comparison of the flotation results indicated that sodium oleate was the more efficient fatty acid for rare earth oxides flotation at all the pulp pH investigated. The flotation performance was found to be particle size-dependent. Both sodium silicate and starch were unselective in decreasing the recovery of iron oxides and silicate minerals, respectively with the corresponding decrease in rare earth oxides recovery. Generally, iron oxides and silicate minerals formed the substantial fraction of the flotation concentrates obtained, both in the absence and presence of depressants, resulting in a generally low rare earth oxides upgrade, even though rare earth oxides recoveries were high. The flotation tests carried out on the tailings sample suggest the feasibility of rare earth oxides recovery using fatty acids, although particle size distribution and minerals liberation are key limiting factors in achieving selective rare earth oxides upgrade.Keywords: depressants, flotation, oleic acid, sodium oleate
Procedia PDF Downloads 1874313 Conditions for Fault Recovery of Interconnected Asynchronous Sequential Machines with State Feedback
Authors: Jung–Min Yang
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In this paper, fault recovery for parallel interconnected asynchronous sequential machines is studied. An adversarial input can infiltrate into one of two submachines comprising parallel composition of the considered asynchronous sequential machine, causing an unauthorized state transition. The control objective is to elucidate the condition for the existence of a corrective controller that makes the closed-loop system immune against any occurrence of adversarial inputs. In particular, an efficient existence condition is presented that does not need the complete modeling of the interconnected asynchronous sequential machine.Keywords: asynchronous sequential machines, parallel composi-tion, corrective control, fault tolerance
Procedia PDF Downloads 2294312 Comparing Double-Stranded RNA Uptake Mechanisms in Dipteran and Lepidopteran Cell Lines
Authors: Nazanin Amanat, Alison Tayler, Steve Whyard
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While chemical insecticides effectively control many insect pests, they also harm many non-target species. Double-stranded RNA (dsRNA) pesticides, in contrast, can be designed to target unique gene sequences and thus act in a species-specific manner. DsRNA insecticides do not, however, work equally well for all insects, and for some species that are considered refractory to dsRNA, a primary factor affecting efficacy is the relative ease by which dsRNA can enter a target cell’s cytoplasm. In this study, we are examining how different structured dsRNAs (linear, hairpin, and paperclip) can enter mosquito and lepidopteran cells, as they represent dsRNA-sensitive and refractory species, respectively. To determine how the dsRNAs enter the cells, we are using chemical inhibitors and RNA interference (RNAi)-mediated knockdown of key proteins associated with different endocytosis processes. Understanding how different dsRNAs enter cells will ultimately help in the design of molecules that overcome refractoriness to RNAi or develop resistance to dsRNA-based insecticides. To date, we have conducted chemical inhibitor experiments on both cell lines and have evidence that linear dsRNAs enter the cells using clathrin-mediated endocytosis, while the paperclip dsRNAs (pcRNAs) can enter both species’ cells in a clathrin-independent manner to induce RNAi. An alternative uptake mechanism for the pcRNAs has been tentatively identified, and the outcomes of our RNAi-mediated knockdown experiments, which should provide corroborative evidence of our initial findings, will be discussed.Keywords: dsRNA, RNAi, uptake, insecticides, dipteran, lepidopteran
Procedia PDF Downloads 724311 The 10,000 Fold Effect Retrograde Neurotransmission: A Newer Concept for Paraplegia’s Physiological Revival by the Use of Intrathecal Sodium Nitroprusside
Authors: V. K. Tewari, M. Hussain, H. K. D. Gupta
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B-Methylprednisolone-level-1-benefit (20%) usually given in paraplegia (but within 8hrs). Patients wait-long-duration for physiological-recovery. Intrathecal-Sodium-Nitroprusside(ITSNP) has been used-in vasospasm-due-to-subarachnoid-hemorrhage. ITSNP-has been studied-here for wide-window-period-range for-treatment, fast-recovery/affordability. 2- for acute-cases-and 1-mechanism-for chronic-cases, which-are-interrelated, are being-proposed-for-physiological-recovery. retrograde-neurotransmission, vasospasm and long-term-potentiation-(ltp) mechanisms are proposed here for recovery. It’s a case-control-prospective-study. 82paraplegia-patients(10patients taken as control-no superfusion or dextrose5% superfusion and 72patients as ITSNP-group). The mean time for superfusion was 14.11 days. ITSNP administered at a dosage of 0.2 mg/kg bo wt. Pre/post ITSNP monitored by SSEP/MEP. After-2-Hours in ITSNP-group Mean-Change-From-Baseline-Asia Motor/Sensory-Score 13.84%/13.10%, after-24-hours MOTOR-1.27-points decrease(3.77%) and SENSORY 10.5points-increase(6.22%)as compared to Control-group no-change noted upto 24-hours, At-7days ITSNP motor/sensory;11.56%/6.22% as compared to Control-group 7.60/4.48%, At-2-months in ITSNP 27.69%/6.22% as compared to Control-group 16.02/4.5%. SSEP/MEP-documented-improvements-noted. ITSNP, a-swift-acting-drug in treatment-of-paraplegia, is effective within-two-hours(mean-change-MOTOR-13.84% and SENSORY-13.10%) on-mean14.11th postparaplegia-day with a small-detrimental-response after-24-hours which-recovers-fast.Keywords: paraplegias, intrathecal sodium nitroprusside, retrograde transmission, the 10, 000 fold effect, perforators, vasodilatations, long term potenciations
Procedia PDF Downloads 4064310 Water Dumpflood into Multiple Low-Pressure Gas Reservoirs
Authors: S. Lertsakulpasuk, S. Athichanagorn
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As depletion-drive gas reservoirs are abandoned when there is insufficient production rate due to pressure depletion, waterflooding has been proposed to increase the reservoir pressure in order to prolong gas production. Due to high cost, water injection may not be economically feasible. Water dumpflood into gas reservoirs is a new promising approach to increase gas recovery by maintaining reservoir pressure with much cheaper costs than conventional waterflooding. Thus, a simulation study of water dumpflood into multiple nearly abandoned or already abandoned thin-bedded gas reservoirs commonly found in the Gulf of Thailand was conducted to demonstrate the advantage of the proposed method and to determine the most suitable operational parameters for reservoirs having different system parameters. A reservoir simulation model consisting of several thin-layered depletion-drive gas reservoirs and an overlying aquifer was constructed in order to investigate the performance of the proposed method. Two producers were initially used to produce gas from the reservoirs. One of them was later converted to a dumpflood well after gas production rate started to decline due to continuous reduction in reservoir pressure. The dumpflood well was used to flow water from the aquifer to increase pressure of the gas reservoir in order to drive gas towards producer. Two main operational parameters which are wellhead pressure of producer and the time to start water dumpflood were investigated to optimize gas recovery for various systems having different gas reservoir dip angles, well spacings, aquifer sizes, and aquifer depths. This simulation study found that water dumpflood can increase gas recovery up to 12% of OGIP depending on operational conditions and system parameters. For the systems having a large aquifer and large distance between wells, it is best to start water dumpflood when the gas rate is still high since the long distance between the gas producer and dumpflood well helps delay water breakthrough at producer. As long as there is no early water breakthrough, the earlier the energy is supplied to the gas reservoirs, the better the gas recovery. On the other hand, for the systems having a small or moderate aquifer size and short distance between the two wells, performing water dumpflood when the rate is close to the economic rate is better because water is more likely to cause an early breakthrough when the distance is short. Water dumpflood into multiple nearly-depleted or depleted gas reservoirs is a novel study. The idea of using water dumpflood to increase gas recovery has been mentioned in the literature but has never been investigated. This detailed study will help a practicing engineer to understand the benefits of such method and can implement it with minimum cost and risk.Keywords: dumpflood, increase gas recovery, low-pressure gas reservoir, multiple gas reservoirs
Procedia PDF Downloads 443