Search results for: chip seal

Authors: Yusser Olguín, Diego Benavente, Fernando Dorta, Nicole Orellana, Cristian Acevedo

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One of the greatest challenges of tissue engineering is the generation of materials in which the highest possible number of conditions can be incorporated to stimulate the proliferation and differentiation of cells, which will be transformed together with the material into new functional tissue. In this sense, considering the properties of microfluidics and its relationship with cellular micro-environments, the possibility of controlling flow patterns and the ability to design diverse patterns in the chips, a microfluidic cell culture system can be established as a means for the evaluation of the effect of different parameters in a controlled and precise manner. Specifically in relation to the study and development of alternatives in peripheral nervous tissue engineering, it is necessary to consider different physical and chemical neurotrophic stimuli that promote cell growth and differentiation. Chemical stimuli include certain vitamins, glucocorticoids, gangliosides, and growth factors, while physical stimuli include topological stimuli, mechanical forces of the cellular environment and electrical stimulation. In this context, the present investigation shows the results of cell stimulation in a microbioreactor using electrical and chemical stimuli, where the differentiation of PC12 cells as a neuronal model is evidenced by neurite expression, dependent on the stimuli and their combination. The results were analysed with a multi-factor statistical approach, showing several relationships and dependencies between different parameters. Chip design, operating parameters and concentrations of neurotrophic chemical factors were found to be preponderant, based on the characteristics of the electrical stimuli.

Keywords: microfluidics, nerve tissue engineering, microbioreactor, electrical stimuli

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Authors: A. Almaneea, N. Kapur, J. L. Summers, H. M. Thompson

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Meeting the growth in demand for digital services such as social media, telecommunications, and business and cloud services requires large scale data centres, which has led to an increase in their end use energy demand. Generally, over 30% of data centre power is consumed by the necessary cooling overhead. Thus energy can be reduced by improving the cooling efficiency. Air and liquid can both be used as cooling media for the data centre. Traditional data centre cooling systems use air, however liquid is recognised as a promising method that can handle the more densely packed data centres. Liquid cooling can be classified into three methods; rack heat exchanger, on-chip heat exchanger and full immersion of the microelectronics. This study quantifies the improvements of heat transfer specifically for the case of immersed microelectronics by varying the CPU and heat sink location. Immersion of the server is achieved by filling the gap between the microelectronics and a water jacket with a dielectric liquid which convects the heat from the CPU to the water jacket on the opposite side. Heat transfer is governed by two physical mechanisms, which is natural convection for the fixed enclosure filled with dielectric liquid and forced convection for the water that is pumped through the water jacket. The model in this study is validated with published numerical and experimental work and shows good agreement with previous work. The results show that the heat transfer performance and Nusselt number (Nu) is improved by 89% by placing the CPU and heat sink on the bottom of the microelectronics enclosure.

Keywords: CPU location, data centre cooling, heat sink in enclosures, immersed microelectronics, turbulent natural convection in enclosures

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Authors: Zakiullah Kalwar, Shabeer Abbassi

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This research encompasses the study of structural styles and evaluation of the hydrocarbon potential of Kotrum and Drigri anticlines located in Rajanpur Area, Midddle Indus Basin of Pakistan with the approach of geophysical data integration. The study area is situated between the Sulaiman Foldbelt on the west and Indus River in the east. It is an anticlinal fold, located to the southeast of Sakhi Sarwar anticline and separated from a prominent syncline. The structure has a narrow elongated crest, with the axis running in SSW-NNE direction. In the east, the structure is bounded by a gentle syncline. Structural Styles are trending East-West and perpendicular to tectonic transport and stress direction and the base of the structures gradually dipping Eastward beneath the deformation frontal part in Eastern Sulaiman Fold Belt. Middle Indus Basin can be divided into Foreland, Sulaiman fold belt and a broad foredeep. Sulaiman represents a blind thrust front, which suggests that all frontal folds of the fold belt are cored by blind thrust. The deformation of frontal part of Sulaiman Lobe represents the passive roof duplex stacked beneath the frontal passive roof thrust. The passive roof thrust, which has a back thrust sense of motion and extends into the interior of Fold belt. Left lateral Kingri Fault separates Eastern and Central Sulaiman fold belt. In Central Sulaiman fold belt the deformation front moved further towards fore deep as compared to Eastern Sulaiman. Two wells (Kotrum-01, Drigri-01) have been drilled in the study area with the objective to determine the potential of oil and gas in Habib Rahi Limestone of Eocene age, Dunghan Limestone of Paleocene age and Pab Sandstone of cretaceous age and role of structural styles in hydrocarbon potential of study area. Kotrum-01 well was drilled to its T.D of 4798m. Besides fishing and side tracking, tight whole conditions, high pressure, and losses of circulation were also encountered. During production, testing Pab sandstone were tested but abandoned found. Drigri-01 well was drilled to its T.D 3250 m. RFT was carried out at different points, but all points showed no pressure / seal failure and the well was plugged and declared abandoned.

Keywords: hydrocarbon potential, structural style, reserve calculation, enhance production

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Authors: Sikiru Oluwarotimi Ismail, Hom Nath Dhakal, Anish Roy, Dong Wang, Ivan Popov

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The ultrasonically-assisted drilling (UAD) is a non-traditional technique which involves the superimposition of a high frequency and low amplitude vibration, usually greater than 18kHz and less than 20µm respectively, on a drill bit along the feed direction. UAD has remarkable advantages over the conventional drilling (CD), especially the high drilling-force reduction. Force reduction improves the quality of the drilled holes, reduces power consumption rate and cost of production. Nevertheless, in addition to the setbacks of UAD including expensiveness of set-up, unpredicted results and chipping effects, this paper presents the problems of insignificant force reduction and poor surface quality during UAD of hemp fibre-reinforced composites (HFRCs), a natural composite, with polycaprolactone (PCL) matrix. The experimental results obtained depict that HFRCs/PCL samples have more burnt chip-materials attached on the drilled holes during UAD than CD. This effect produced a very high surface roughness (Ra), up to 13µm. In a bid to reduce these challenges, different drilling parameters (feed rates and cutting speeds, frequencies and amplitudes for UAD), conditions (dry machining and airflow cooling) and diameters of drill bits (3mm and 6mm of high speed steel), as well as HFRCs/PCL samples of various fibre aspect ratios, including 0 (neat), 19, 26, 30 and 38 have been used. However, the setbacks still persisted. Evidently, the benefits of UAD are not obtainable for the drilling of the HFRCs/PCL laminates. These problems occurred due to the 60 °C melting temperature of PCL, quite lower than 56-90.2 °C and 265–290.8 °C composite-tool interface temperature during CD and UAD respectively.

Keywords: force reduction, hemp fibre-reinforced composites, ultrasonically-assisted drilling, surface quality

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Authors: Amina Farooq, Nauman Zafar Butt

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This paper is about methods and tools for counting particles of interest, such as cells. A microfluidic system with interconnected electronics on a flexible substrate, inlet-outlet ports and interface schemes, sensitive and selective detection of cells specificity, and processing of cell counting at polymer interfaces in a microscale biosensor for use in the detection of target biological and non-biological cells. The development of fluidic channels, planar fluidic contact ports, integrated metal electrodes on a flexible substrate for impedance measurements, and a surface modification plasma treatment as an intermediate bonding layer are all part of the fabrication process. Magnetron DC sputtering is used to deposit a double metal layer (Ti/Pt) over the polypropylene film. Using a photoresist layer, specified and etched zones are established. Small fluid volumes, a reduced detection region, and electrical impedance measurements over a range of frequencies for cell counts improve detection sensitivity and specificity. The procedure involves continuous flow of fluid samples that contain particles of interest through the microfluidic channels, counting all types of particles in a portion of the sample using the electrical differential counter to generate a bipolar pulse for each passing cell—calculating the total number of particles of interest originally in the fluid sample by using MATLAB program and signal processing. It's indeed potential to develop a robust and economical kit for cell counting in whole-blood samples using these methods and similar devices.

Keywords: impedance, biochip, cell counting, microfluidics

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Authors: Fatma Dridi, Mouna Marrakchi, Mohammed Gargouri, Alvaro Garcia Cruz, Sergei V. Dzyadevych, Francis Vocanson, Joëlle Saulnier, Nicole Jaffrezic-Renault, Florence Lagarde

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An original method has been successfully developed for the immobilization of thermolysin onto gold interdigitated electrodes for the detection of ochratoxin A (OTA) in olive oil samples. A mix of polyvinyl alcohol (PVA), polyethylenimine (PEI) and gold nanoparticles (AuNPs) was used. Cross-linking sensors chip was made by using a saturated glutaraldehyde (GA) vapor atmosphere in order to render the two polymers water stable. Performance of AuNPs/ (PVA/PEI) modified electrode was compared to a traditional immobilized enzymatic method using bovine serum albumin (BSA). Atomic force microscopy (AFM) experiments were employed to provide a useful insight into the structure and morphology of the immobilized thermolysin composite membranes. The enzyme immobilization method influence the topography and the texture of the deposited layer. Biosensors optimization and analytical characteristics properties were studied. Under optimal conditions AuNPs/ (PVA/PEI) modified electrode showed a higher increment in sensitivity. A 700 enhancement factor could be achieved with a detection limit of 1 nM. The newly designed OTA biosensors showed a long-term stability and good reproducibility. The relevance of the method was evaluated using commercial doped olive oil samples. No pretreatment of the sample was needed for testing and no matrix effect was observed. Recovery values were close to 100% demonstrating the suitability of the proposed method for OTA screening in olive oil.

Keywords: thermolysin, A. ochratoxin , polyvinyl alcohol, polyethylenimine, gold nanoparticles, olive oil

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Authors: Aubry Martin, Nehed Amara, Jeff Nyalosaso, Audrey Potdevin, FrançOis ReVeret, Michel Langlet, Genevieve Chadeyron

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Energy-saving lighting devices based on LightEmitting Diodes (LEDs) combine a semiconductor chip emitting in the ultraviolet or blue wavelength region to one or more phosphor(s) deposited in the form of coatings. The most common ones combine a blue LED with the yellow phosphor Y₃Al₅O₁₂:Ce³⁺ (YAG:Ce) and a red phosphor. Even if these devices are characterized by satisfying photometric parameters (Color Rendering Index, Color Temperature) and good luminous efficiencies, further improvements can be carried out to enhance light extraction efficiency (increase in phosphor forward emission). One of the possible strategies is to pattern the phosphor coatings. Here, we have worked on different ways to nanostructure the coating surface. On the one hand, we used the colloidal lithography combined with the Langmuir-Blodgett technique to directly pattern the surface of YAG:Tb³⁺ sol-gel derived coatings, YAG:Tb³⁺ being used as phosphor model. On the other hand, we achieved composite architectures combining YAG:Ce coatings and ZnO nanowires. Structural, morphological and optical properties of both systems have been studied and compared to flat YAG coatings. In both cases, nanostructuring brought a significative enhancement of photoluminescence properties under UV or blue radiations. In particular, angle-resolved photoluminescence measurements have shown that nanostructuring modifies photons path within the coatings, with a better extraction of the guided modes. These two strategies have the advantage of being versatile and applicable to any phosphor synthesizable by sol-gel technique. They then appear as promising ways to enhancement luminescence efficiencies of both phosphor coatings and the optical devices into which they are incorporated, such as LED-based lighting or safety devices.

Keywords: phosphor coatings, nanostructuring, light extraction, ZnO nanowires, colloidal lithography, LED devices

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Authors: P. van Tonder, U. Bagdadi, B. M. D. Lario, Z. Masina, T. R. Motshwari

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This paper analyses how concrete reinforcing steel bars corrode and how it can be minimised through the use of various protection methods against corrosion, such as metal-based paint, alloying, cathodic protection and electroplating. Samples of carbon steel bars were protected, using these four methods. Tests performed on the samples included durability, electrical resistivity and bond strength. Durability results indicated relatively low corrosion rates for alloying, cathodic protection, electroplating and metal-based paint. The resistivity results indicate all samples experienced a downward trend, despite erratic fluctuations in the data, indicating an inverse relationship between electrical resistivity and corrosion rate. The results indicated lowered bond strengths when the reinforced concrete was cured in seawater compared to being cured in normal water. It also showed that higher design compressive strengths lead to higher bond strengths which can be used to compensate for the loss of bond strength due to corrosion in a real-world application. In terms of implications, all protection methods have the potential to be effective at resisting corrosion in real-world applications, especially the alloying, cathodic protection and electroplating methods. The metal-based paint underperformed by comparison, most likely due to the nature of paint in general which can fade and chip away, revealing the steel samples and exposing them to corrosion. For alloying, stainless steel is the suggested material of choice, where Y-bars are highly recommended as smooth bars have a much-lowered bond strength. Cathodic protection performed the best of all in protecting the sample from corrosion, however, its real-world application would require significant evaluation into the feasibility of such a method.

Keywords: protection methods, corrosion, concrete, reinforcing steel bars

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Authors: Radka Obořilová, Hana Šimečková, Matěj Pastucha, Jan Přibyl, Petr Skládal, Ivana Mašlaňová, Zdeněk Farka

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Due to the spreading of antimicrobial resistance, alternative approaches to combat superinfections are being sought, both in the field of lysing agents and methods for studying bacterial lysis. A suitable alternative to antibiotics is phage therapy and enzybiotics, for which it is also necessary to study the mechanism of their action. Biosensor-based techniques allow rapid detection of pathogens in real time, verification of sensitivity to commonly used antimicrobial agents, and selection of suitable lysis agents. The detection of lysis takes place on the surface of the biosensor with immobilized bacteria, which has the potential to be used to study biofilms. An example of such a biosensor is surface plasmon resonance (SPR), which records the kinetics of bacterial lysis based on a change in the resonance angle. The bacteria are immobilized on the surface of the SPR chip, and the action of phage as the mass loss is monitored after a typical lytic cycle delay. Atomic force microscopy (AFM) is a technique for imaging of samples on the surface. In contrast to electron microscopy, it has the advantage of real-time imaging in the native conditions of the nutrient medium. In our case, Staphylococcus aureus was lysed using the enzyme lysostaphin and phage P68 from the familyPodoviridae at 37 ° C. In addition to visualization, AFM was used to study changes in mechanical properties during lysis, which resulted in a reduction of Young’s modulus (E) after disruption of the bacterial wall. Changes in E reflect the stiffness of the bacterium. These advanced methods provide deeper insight into bacterial lysis and can help to fight against bacterial diseases.

Keywords: biosensors, atomic force microscopy, surface plasmon resonance, bacterial lysis, staphylococcus aureus, phage P68

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Authors: Yuxiang Pan, Yong Qiu, Chenlei Gu, Ping Wang

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In the past decade, three-dimensional (3D) tumor cell models have attracted increasing interest in the field of drug screening due to their great advantages in simulating more accurately the heterogeneous tumor behavior in vivo. Drug sensitivity testing based on 3D tumor cell models can provide more reliable in vivo efficacy prediction. The gold standard fluorescence staining is hard to achieve the real-time and label-free monitoring of the viability of 3D tumor cell models. In this study, micro-groove impedance sensor (MGIS) was specially developed for dynamic and non-invasive monitoring of 3D cell viability. 3D tumor cells were trapped in the micro-grooves with opposite gold electrodes for the in-situ impedance measurement. The change of live cell number would cause inversely proportional change to the impedance magnitude of the entire cell/matrigel to construct and reflect the proliferation and apoptosis of 3D cells. It was confirmed that 3D cell viability detected by the MGIS platform is highly consistent with the standard live/dead staining. Furthermore, the accuracy of MGIS platform was demonstrated quantitatively using 3D lung cancer model and sophisticated drug sensitivity testing. In addition, the parameters of micro-groove impedance chip processing and measurement experiments were optimized in details. The results demonstrated that the MGIS and 3D cell-based biosensor and would be a promising platform to improve the efficiency and accuracy of cell-based anti-cancer drug screening in vitro.

Keywords: micro-groove impedance sensor, 3D cell-based biosensors, 3D cell viability, micro-electromechanical systems

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Authors: Iman Kamranfar, Gang-Ping Xue, Salma Balazadeh, Bernd Mueller-Roeber

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Adverse environmental conditions such as salinity stress, high temperature and drought limit plant growth and typically lead to precocious tissue degeneration and leaf senescence, a process by which nutrients from photosynthetic organs are recycled for the formation of flowers and seeds to secure reaching the next generation under such harmful conditions. In addition, abiotic stress affects developmental patterns that help the plant to withstand unfavourable environmental conditions. We discovered an NAC (for NAM, ATAF1, 2, and CUC2) transcription factor (TF), called RAF in the following, which plays a central role in abiotic drought stress-triggered senescence and the control of developmental adaptations to stressful environments. RAF is an ABA-responsive TF; RAF overexpressors are hypersensitive to abscisic acid (ABA) and exhibit precocious senescence while knock-out mutants show delayed senescence. To explore the RAF gene regulatory network (GRN), we determined its preferred DNA binding sites by binding site selection assay (BSSA) and performed microarray-based expression profiling using inducible RAF overexpression lines and chromatin immunoprecipitation (ChIP)-PCR. Our studies identified several direct target genes, including those encoding for catabolic enzymes acting during stress-induced senescence. Furthermore, we identified various genes controlling drought stress-related developmental changes. Based on our results, we conclude that RAF functions as a central transcriptional regulator that coordinates developmental programs with stress-related inputs from the environment. To explore the potential agricultural applications of our findings, we are currently extending our studies towards crop species.

Keywords: abiotic stress, Arabidopsis, development, transcription factor

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Authors: Ruth Lorivi, Jianzheng Li, John J. Ambuchi, Kaiwen Deng

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Nitrogen removal from wastewater is accomplished by nitrification and denitrification processes. Successful denitrification requires carbon, therefore, if placed after biochemical oxygen demand (BOD) and nitrification process, a carbon source has to be re-introduced into the water. To avoid adding a carbon source, denitrification is usually placed before BOD and nitrification processes. This process however involves recycling the nitrified effluent. In this study wood chips were used as internal carbon source which enabled placement of denitrification after BOD and nitrification process without effluent recycling. To investigate the efficiency of a wood packed aerobic-anaerobic baffled reactor on carbon and nutrients removal from domestic wastewater, a three compartment baffled reactor was presented. Each of the three compartments was packed with 329 g wood chips 1x1cm acting as an internal carbon source for denitrification. The proposed mode of operation was aerobic-anoxic-anaerobic (OAA) with no effluent recycling. The operating temperature, hydraulic retention time (HRT), dissolved oxygen (DO) and pH were 24 ± 2 ℃, 24 h, less than 4 mg/L and 7 ± 1 respectively. The removal efficiencies of chemical oxygen demand (COD), ammonia nitrogen (NH₄⁺-N) and total nitrogen (TN) attained was 99, 87 and 83% respectively. TN removal rate was limited by nitrification as 97% of ammonia converted into nitrate and nitrite was denitrified. These results show that application of wood chips in wastewater treatment processes is an efficient internal carbon source. 

Keywords: aerobic-anaerobic baffled reactor, denitrification, nitrification, wood chip

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Authors: Gaurav D. Sonawane, Vikas G. Sargade

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The present investigation is a study of the effect of advanced Physical Vapor Deposition (PVD) coatings on cutting temperature residual stresses and surface roughness during Duplex Stainless Steel (DSS) 2205 turning. Austenite stabilizers like nickel, manganese, and molybdenum reduced the cost of DSS. Surface Integrity (SI) plays an important role in determining corrosion resistance and fatigue life. Resistance to various types of corrosion makes DSS suitable for applications with critical environments like Heat exchangers, Desalination plants, Seawater pipes and Marine components. However, lower thermal conductivity, poor chip control and non-uniform tool wear make DSS very difficult to machine. Cemented carbide tools (M grade) were used to turn DSS in a dry environment. AlTiN and AlTiCrN coatings were deposited using advanced PVD High Pulse Impulse Magnetron Sputtering (HiPIMS) technique. Experiments were conducted with cutting speed of 100 m/min, 140 m/min and 180 m/min. A constant feed and depth of cut of 0.18 mm/rev and 0.8 mm were used, respectively. AlTiCrN coated tools followed by AlTiN coated tools outperformed uncoated tools due to properties like lower thermal conductivity, higher adhesion strength and hardness. Residual stresses were found to be compressive for all the tools used for dry turning, increasing the fatigue life of the machined component. Higher cutting temperatures were observed for coated tools due to its lower thermal conductivity, which results in very less tool wear than uncoated tools. Surface roughness with uncoated tools was found to be three times higher than coated tools due to lower coefficient of friction of coating used.

Keywords: cutting temperature, DSS2205, dry turning, HiPIMS, surface integrity

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Authors: German Ricci

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The research aims to analyse the symbolic power of the International Monetary Fund (IMF) in its relationship with a borrowing country, drawing upon Pierre Bourdieu’s Field Theory. This theory of power, typical of constructivist structuralism, has been minor used in international relations. Thus, selecting this perspective offers a new understanding of how the IMF's power operates and is structured. The IMF makes periodic economic reviews in which the staff evaluates the Government's performance. It also offers “last instance” loans when private external credit is not accessible. This relationship generates great expectations in financial agents because the IMF’s statements indicate the capacity of the Nation-State to meet its payment obligations (or not). Therefore, it is argued that the IMF is a legitimate actor for financial agents concerned about a government facing an economic crisis both for the effects of its immediate economic contribution through loans and the promotion of adjustment programs, helpful to guarantee the payment of the external debt. This legitimacy implies a symbolic power relationship in addition to the already known economic power relationship. Obtaining the IMF's consent implies that the government partially puts its political-economic decisions into play since the monetary policy must be agreed upon with the Fund. This has consequences at the local level. First, it implies that the debtor state must establish a daily relationship with the Fund. This everyday interaction with the Fund influences how officials and policymakers internalize the meaning of political management. On the other hand, if the Government has access to the IMF's seal of approval, the State will be again in a position to re-enter the financial market and go back into debt to face external debt. This means that private creditors increase the chances of collecting the debt and, again, grant credits. Thus, it is argued that the borrowing country submits to the relationship with the IMF in search of the latter's economic and symbolic capital. Access to this symbolic capital has objective and subjective repercussions at the national level that might tend to reproduce the relevance of the financial market and legitimizes the IMF’s intervention during economic crises. The paper has Argentina as its case study, given its historical relationship with the IMF and the relevance of the current indebtedness period, which remains largely unexplored. Argentina’s economy is characterized by recurrent financial crises, and it is the country to which the Fund has lent the most in its entire history. It surpasses more than three times the second, Egypt. In addition, Argentina is currently the country that owes the most to the Fund after receiving the largest loan ever granted by the IMF in 2018, and a new agreement in 2022. While the historical strong association with the Fund culminated in the most acute economic and social crisis in the country’s contemporary history, producing an unprecedented political and institutional crisis in 2001, Argentina still recognized the IMF as the only way out during economic crises.

Keywords: IMF, fields theory, symbolic power, Argentina, Bourdieu

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Authors: Binod C. Agrawal

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In modern history, industrial and media merchandising in South Asia from East Asia, Europe, United States and other countries of the West is over 200 years old. Hence, continued external technology and media exposure is not a new experience in multi-lingual and multi religious South Asia which evolved cultural means to withstand structural change. In the post-World War II phase, media exposure especially of telecommunication, film, Internet, radio, print media and television have increased manifold. South Asia did not lose any time in acquiring and adopting digital media accelerated by chip revolution, computer and satellite communication. The penetration of digital media and utilization are exceptionally high though the spread has an unequal intensity, use and effects. The author argues that industrial and media products are “cultural products” apart from being “technological products”; hence their influences are most felt in the cultural domain which may lead to blunting of unique cultural specifics in the multi-cultural, multi-lingual and multi religious South Asia. Social scientists, political leaders and parents have voiced concern of “Cultural domination”, “Digital media colonization” and “Westernization”. Increased digital media access has also opened up doors of pornography and other harmful information that have sparked fresh debates and discussions about serious negative, harmful, and undesirable social effects especially among youth. Within ‘techno-social’ perspective, based on recent research studies, the paper aims to describe and analyse possible socio-economic change due to digital media penetration. Further, analysis supports the view that the ancient multi-lingual and multi-religious cultures of South Asia due to inner cultural strength may sustain without setting in a process of irreversible structural changes in South Asia.

Keywords: cultural sustainability, digital media effects, digital media impact in South Asia, social change in South Asia

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Authors: Fahad Qureshi

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This paper provides detailed study on the diagnosis of intermittent high vibration spikes at exhaust bearing (Non-Drive End) of Mitsubishi H-25 gas turbine installed in a petrochemical plant in Pakistan. The diagnosis is followed by successful root cause analysis of the issue and recommendations for improving the reliability of machine. Engro Polymer and Chemicals (EPCL), a Chlor Vinyl complex, has a captive power plant consisting of one combined cycle power plant (CCPP), having two gas turbines each having 25 MW capacity (make: Hitachi) and one extraction condensing steam turbine having 15 MW capacity (make: HTC). Besides, one 6.75 MW SGT-200 1S gas turbine (make: Alstom) is also available. In 2018, the organization faced an issue of intermittent high vibration at exhaust bearing of one of H-25 units having tag GT-2101 A, which eventually led to tripping of machine at configured securities. Since the machine had surpassed 64,000 running hours and major inspection was also due, so bearings inspection was performed. Inspection revealed excessive coke deposition at labyrinth where evidence of rotor rub was also present. Bearing clearance was also at upper limit, and slight babbitt (soft metal) chip off was observed at one of its pads so it was preventively replaced. The unit was restated successfully and exhibited no abnormality until October 2020, when these spikes reoccurred, leading to machine trip. Recurrence of the issue within two years indicated that root cause was not properly addressed, so this paper furthers the discussion on in-depth analysis of findings and establishes successful root cause analysis, which captured significant learnings both in terms of machine design deficiencies and gaps in operation & maintenance (O & M) regime. Lastly, revised O& M regime along with set of recommendations are proposed to avoid recurrence.

Keywords: exhaust side bearing, Gas turbine, rubbing, vibration

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Authors: Sneha Kumari, Ravi Krishnan Elangovan

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This abstract describes the development of a high-speed tracking method by modification in motor components for nanoparticle attachment. Myosin motors are nano-sized protein machines powering movement that defines life. These miniature molecular devices serve as engines utilizing chemical energy stored in ATP to produce useful mechanical energy in the form of a few nanometre displacement events leading to force generation that is required for cargo transport, cell division, cell locomotion, translated to macroscopic movements like running etc. With the advent of in vitro motility assay (IVMA), detailed functional studies of the actomyosin system could be performed. The major challenge with the currently available IVMA for tracking actin filaments is a resolution limitation of ± 50nm. To overcome this, we are trying to develop Single Molecule IVMA in which nanoparticle (GNP/QD) will be attached along or on the barbed end of actin filaments using CapZ protein and visualization by a compact TIRF module called ‘cTIRF’. The waveguide-based illumination by cTIRF offers a unique separation of excitation and collection optics, enabling imaging by scattering without emission filters. So, this technology is well equipped to perform tracking with high precision in temporal resolution of 2ms with significantly improved SNR by 100-fold as compared to conventional TIRF. Also, the nanoparticles (QD/GNP) attached to actin filament act as a point source of light coffering ease in filament tracking compared to conventional manual tracking. Moreover, the attachment of cargo (QD/GNP) to the thin filament paves the way for various nano-technological applications through their transportation to different predetermined locations on the chip

Keywords: actin, cargo, IVMA, myosin motors and single-molecule system

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Authors: Graham Hancox, Saydia Razak, Sue Hignett, Jo Barnes, Jyri Silmari, Florian Kading

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In the event of a Chemical, Biological, Radiological and Nuclear (CBRN) incident rapidly gaining, situational awareness is of paramount importance and advanced technologies have an important role to play in improving detection, identification, monitoring (DIM) and patient tracking. Understanding how these advanced technologies can fit into current response systems is essential to ensure they are optimally designed, usable and meet end-users’ needs. For this reason, Human Factors (Ergonomics) methods have been used within an EU Horizon 2020 project (TOXI-Triage) to firstly describe (map) the hierarchical structure in a CBRN response with adapted Accident Map (AcciMap) methodology. Secondly, Hierarchical Task Analysis (HTA) has been used to describe and review the sequence of steps (sub-tasks) in a CBRN scenario response as a task system. HTA methodology was then used to map one advanced technology, ‘Tag and Trace’, which tags an element (people, sample and equipment) with a Near Field Communication (NFC) chip in the Hot Zone to allow tracing of (monitoring), for example casualty progress through the response. This HTA mapping of the Tag and Trace system showed how the provider envisaged the technology being used, allowing for review and fit with the current CBRN response systems. These methodologies have been found to be very effective in promoting and supporting a dialogue between end-users and technology providers. The Human Factors methods have given clear diagrammatic (visual) representations of how providers see their technology being used and how end users would actually use it in the field; allowing for a more user centered approach to the design process. For CBRN events usability is critical as sub-optimum design of technology could add to a responders’ workload in what is already a chaotic, ambiguous and safety critical environment.

Keywords: AcciMap, CBRN, ergonomics, hierarchical task analysis, human factors

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Authors: Meng-Jie Hsiao, Cam Nguyen

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Wide-band devices in Ku band (12-18 GHz) and K band (18-27 GHz) have received significant attention for high-data-rate communications and high-resolution sensing. Especially, devices operating around 24 GHz is attractive due to the 24-GHz unlicensed applications. One of the most important components in RF systems is power amplifier (PA). Various PAs have been developed in the Ku and K bands on GaAs, InP, and silicon (Si) processes. Although the PAs using GaAs or InP process could have better power handling and efficiency than those realized on Si, it is very hard to integrate the entire system on the same substrate for GaAs or InP. Si, on the other hand, facilitates single-chip systems. Hence, good PAs on Si substrate are desirable. Especially, Si-based PA having good linearity is necessary for next generation communication protocols implemented on Si. We report a 16.5 to 25.5 GHz Si-based PA having flat saturated power of 19.5 ± 1.5 dBm, output 1-dB power compression (OP1dB) of 16.5 ± 1.5 dBm, and 15-23 % power added efficiency (PAE). The PA consists of a drive amplifier, two main amplifiers, and lump-element Wilkinson power divider and combiner designed and fabricated in TowerJazz 0.18µm SiGe BiCMOS process having unity power gain frequency (fMAX) of more than 250 GHz. The PA is realized as a cascode amplifier implementing both heterojunction bipolar transistor (HBT) and n-channel metal–oxide–semiconductor field-effect transistor (NMOS) devices for gain, frequency response, and linearity consideration. Particularly, a body-floating technique is utilized for the NMOS devices to improve the voltage swing and eliminate parasitic capacitances. The developed PA has measured flat gain of 20 ± 1.5 dB across 16.5-25.5 GHz. At 24 GHz, the saturated power, OP1dB, and maximum PAE are 20.8 dBm, 18.1 dBm, and 23%, respectively. Its high performance makes it attractive for use in Ku/K-band, especially 24 GHz, communication and radar systems. This paper was made possible by NPRP grant # 6-241-2-102 from the Qatar National Research Fund (a member of Qatar Foundation). The statements made herein are solely the responsibility of the authors.

Keywords: power amplifiers, amplifiers, communication systems, radar systems

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Authors: Udayan Patankar, Shashwati Bhagat, Vilas Nitneware, Ants Koel

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An ISM band power amplifier is a type of electronic amplifier used to convert a low-power radio-frequency signal into a larger signal of significant power, typically used for driving the antenna of a transmitter. Due to drastic changes in telecommunication generations may lead to the requirements of improvements. Rapid changes in communication lead to the wide implementation of WLAN technology for its excellent characteristics, such as high transmission speed, long communication distance, and high reliability. Many applications such as WLAN, Bluetooth, and ZigBee, etc. were evolved with 2.4GHz to 5 GHz ISM Band, in which the power amplifier (PA) is a key building block of RF transmitters. There are many manufacturing processes available to manufacture a power amplifier for desired power output, but the major problem they have faced is about the power it consumed for its proper working, as many of them are fabricated on the GaN HEMT, Bi COMS process. In this paper we present a CMOS Base two stage cascode design of power amplifier working on 2.4GHz ISM frequency band. To lower the costs and allow full integration of a complete System-on-Chip (SoC) we have chosen 0.13µm low power CMOS technology for design. While designing a power amplifier, it is a real task to achieve higher power efficiency with minimum resources. This design showcase the Multi biased Cascode methodology to implement a two-stage CMOS power amplifier using ADS and LTSpice simulating tool. Main source is maximum of 2.4V which is internally distributed into different biasing point VB driving and VB driven as required for distinct stages of two stage RF power amplifier. It shows maximum power added efficiency near about 70.195% whereas its Power added efficiency calculated at 1 dB compression point is 44.669 %. Biased MOSFET is used to reduce total dc current as this circuit is designed for different wireless applications comes under 2.4GHz ISM Band.

Keywords: RFIC, PAE, RF CMOS, impedance matching

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Authors: Jiping Cai, Ningzhi Wangyang, Jun Shao

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Traumatic brain injury (TBI) is one of the major causes of morbidity and mortality that leads to structural and functional damage in several parts of the brain, such as cranial nerves, optic nerve tract or other circuitry involved in vision and occipital lobe, depending on its location and severity. As a result, the function associated with vision processing and perception are significantly affected and cause blurred vision, double vision, decreased peripheral vision and blindness. Here two cases complaining of monocular vision loss (actually temporal hemianopia) due to traumatic chiasmal syndrome after frontal head injury were reported, and were compared the findings with individual case reports published in the literature. Reported cases of traumatic chiasmal syndrome appear to share some common features, such as injury to the frontal bone and fracture of the anterior skull base. The degree of bitemporal hemianopia and visual loss acuity have a variable presentation and was not necessarily related to the severity of the craniocerebral trauma. Chiasmal injury may occur even in the absence bony chip impingement. Isolated bitemporal hemianopia is rare and clinical improvement usually may not occur. Mechanisms of damage to the optic chiasm after trauma include direct tearing, contusion haemorrhage and contusion necrosis, and secondary mechanisms such as cell death, inflammation, edema, neurogenesis impairment and axonal damage associated with TBI. Beside visual field test, MRI evaluation of optic pathways seems to the strong objective evidence to demonstrate the impairment of the integrity of visual systems following TBI. Therefore, traumatic chiasmal syndrome should be considered as a differential diagnosis by both neurosurgeons and ophthalmologists in patients presenting with visual impairment, especially bitemporal hemianopia after head injury causing frontal and anterior skull base fracture.

Keywords: bitemporal hemianopia, brain injury, optic chiasma, traumatic chiasmal syndrome.

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Authors: Yongyi Chen, Li Qin, Peng Jia, Yongqiang Ning, Yun Liu, Lijun Wang

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The distributed feedback (DFB) lasers are indispensable in optical phase array (OPA) used for light detection and ranging (LIDAR) techniques, laser communication systems and integrated optics, thanks to their stable single longitudinal mode and narrow linewidth properties. Traditional index-coupled (IC) DFB lasers with uniform gratings have an inherent problem of lasing two degenerated modes. Phase shifts are usually required to eliminate the mode degeneration, making the grating structure complex and expensive. High-quality antireflection (AR) coatings on both lasing facets are also essential owing to the random facet phases introduced by the chip cleavage process, which means half of the lasing energy is wasted. Gain-coupled DFB (GC-DFB) lasers based on the periodic gain (or loss) are announced to have single longitudinal mode as well as capable of the unsymmetrical coating to increase lasing power and efficiency thanks to facet immunity. However, expensive and time-consuming technologies such as epitaxial regrowth and nanoscale grating processing are still required just as IC-DFB lasers, preventing them from practical applications and commercial markets. In this research, we propose a low-cost, single-mode regrowth-free GC-DFB laser based on periodic surface p-contacts. The gain coupling effect is achieved simply by periodic current distribution in the quantum well caused by periodic surface p-contacts, introducing very little index-coupling effect that can be omitted. It is prepared by i-line lithography, without nanoscale grating fabrication or secondary epitaxy. Due to easy fabrication techniques, it provides a method to fabricate practical low cost GC-DFB lasers for widespread practical applications.

Keywords: DFB laser, gain-coupled, low cost, periodic p-contacts

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Authors: Ahmad Rasyadan Arshad, A. Rahman A. Jamal, N. Mohamed Ibrahim, Nor Azian Abdul Murad

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Introduction: Parkinson’s disease (PD) is a complex and asymptomatic disease where patients are usually diagnosed at late stage where about 70% of the dopaminergic neurons are lost. Therefore, identification of molecular biomarkers is crucial for early diagnosis of PD. MicroRNA (miRNA) is a short nucleotide non-coding small RNA which regulates the gene expression in post-translational process. The involvement of these miRNAs in neurodegenerative diseases includes maintenance of neuronal development, necrosis, mitochondrial dysfunction and oxidative stress. Thus, miRNA could be a potential biomarkers for diagnosis of PD. Objective: This study aim to identify the miRNA involved in Late Onset PD (LOPD) and Early Onset PD (EOPD) compared to the controls. Methods: This is a case-control study involved PD patients in the Chancellor Tunku Muhriz Hospital at the UKM Medical Centre. miRNA samples were extracted using miRNeasy serum/plasma kit from Qiagen. The quality of miRNA extracted was determined using Agilent RNA 6000 Nano kit in the Bioanalyzer. miRNA expression was performed using GeneChip miRNA 4.0 chip from Affymetrix. Microarray was performed in EOPD (n= 7), LOPD (n=9) and healthy control (n=11). Expression Console and Transcriptomic Analyses Console were used to analyze the microarray data. Result: miR-129-5p was significantly downregulated in EOPD compared to LOPD with -4.2 fold change (p = <0.050. miR-301a-3p was upregulated in EOPD compared to healthy control (fold = 10.3, p = <0.05). In LOPD versus healthy control, miR-486-3p (fold = 15.28, p = <0.05), miR-29c-3p (fold = 12.21, p = <0.05) and miR-301a-3p (fold = 10.01, p =< 0.05) were upregulated. Conclusion: Several miRNA have been identified to be differentially expressed in EOPD compared to LOPD and PD versus control. These miRNAs could serve as the potential biomarkers for early diagnosis of PD. However, these miRNAs need to be validated in a larger sample size.

Keywords: early onset PD, late onset PD, microRNA (miRNA), microarray

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Authors: Hamed Movahedi, Nicolas Bovet, Henning Friis Poulsen

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Following the advent of the Industrial Revolution, there has been a significant increase in the extraction and utilization of hydrocarbon and fossil fuel resources. However, a new era has emerged, characterized by a shift towards sustainable practices, namely the reduction of carbon emissions and the promotion of renewable energy generation. Given the substantial number of mature oil and gas wells that have been developed inside the petroleum reservoir domain, it is imperative to establish an environmental strategy and adopt appropriate measures to effectively seal and decommission these wells. In general, the cement plug serves as a material for plugging purposes. Nevertheless, there exist some scenarios in which the durability of such a plug is compromised, leading to the potential escape of hydrocarbons via fissures and fractures within cement plugs. Furthermore, cement is often not considered a practical solution for temporary plugging, particularly in the case of well sites that have the potential for future gas storage or CO2 injection. The Danish oil and gas industry has promising potential as a prospective candidate for future carbon dioxide (CO2) injection, hence contributing to the implementation of carbon capture strategies within Europe. The primary reservoir component consists of chalk, a rock characterized by limited permeability. This work focuses on the development and characterization of a novel hydrogel variant. The hydrogel is designed to be injected via a low-permeability reservoir and afterward undergoes a transformation into a high-viscosity gel. The primary objective of this research is to explore the potential of this hydrogel as a new solution for effectively plugging well flow. Initially, the synthesis of polyacrylamide was carried out using radical polymerization inside the confines of the reaction flask. Subsequently, with the application of the Hoffman rearrangement, the polymer chain undergoes partial amination, facilitating its subsequent reaction with the crosslinker and enabling the formation of a hydrogel in the subsequent stage. The organic crosslinker, glutaraldehyde, was employed in the experiment to facilitate the formation of a gel. This gel formation occurred when the polymeric solution was subjected to heat within a specified range of reservoir temperatures. Additionally, a rheological survey and gel time measurements were conducted on several polymeric solutions to determine the optimal concentration. The findings indicate that the gel duration is contingent upon the starting concentration and exhibits a range of 4 to 20 hours, hence allowing for manipulation to accommodate diverse injection strategies. Moreover, the findings indicate that the gel may be generated in environments characterized by acidity and high salinity. This property ensures the suitability of this substance for application in challenging reservoir conditions. The rheological investigation indicates that the polymeric solution exhibits the characteristics of a Herschel-Bulkley fluid with somewhat elevated yield stress prior to solidification.

Keywords: polyacrylamide, hofmann rearrangement, rheology, gel time

Procedia PDF Downloads 52

Authors: P. Bauer, K. Mougin, V. Vignal, A. Buch, P. Ponthiaux, D. Faye

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Nowadays, noble metallic nanostructures with unique morphology are widely used as new sensors due to their fascinating optical, electronic and catalytic properties. Among various shapes, dendritic nanostructures have attracted much attention because of their large surface-to-volume ratio, high sensitivity and special texture with sharp tips and nanoscale junctions. Several methods have been developed to fabricate those specific structures such as electrodeposition, photochemical way, seed-mediated growth or wet chemical method. The present study deals with a novel approach for a controlled growth pattern-directed organisation of Au flower-like crystals (NFs) deposited onto stainless steel plates to achieve large-scale functional surfaces. This technique consists in the deposition of a soft nanoporous template on which Au NFs are grown by electroplating and seed-mediated method. Size, morphology, and interstructure distance have been controlled by a site selective nucleation process. Dendritic Au nanostructures have appeared as excellent Raman-active candidates due to the presence of very sharp tips of multi-branched Au nanoparticles that leads to a large local field enhancement and a good SERS sensitivity. In addition, these structures have also been used as electrochemical sensors to detect traces of molecules present in a solution. A correlation of the number of active sites on the surface and the current charge by both colorimetric method and cyclic voltammetry of gold structures have allowed a calibration of the system. This device represents a first step for the fabrication of MEMs platform that could ultimately be integrated into a lab-on-chip system. It also opens pathways to several technologically large-scale nanomaterials fabrication such as hierarchically ordered crystal architectures for sensor applications.

Keywords: dendritic, electroplating, gold, template

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Authors: Vishal Pareek, Shubham Gupta, Sushil Chandra Jain

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With increasing high-speed computation demand the power consumption, heat dissipation and chip size issues are posing challenges for logic design with conventional technologies. Recovery of bit loss and bit errors is other issues that require reversibility and fault tolerance in the computation. The reversible computing is emerging as an alternative to conventional technologies to overcome the above problems and helpful in a diverse area such as low-power design, nanotechnology, quantum computing. Bit loss issue can be solved through unique input-output mapping which require reversibility and bit error issue require the capability of fault tolerance in design. In order to incorporate reversibility a number of combinational reversible logic based circuits have been developed. However, very few sequential reversible circuits have been reported in the literature. To make the circuit fault tolerant, a number of fault model and test approaches have been proposed for reversible logic. In this paper, we have attempted to incorporate fault tolerance in sequential reversible building blocks such as D flip-flop, T flip-flop, JK flip-flop, R-S flip-flop, Master-Slave D flip-flop, and double edge triggered D flip-flop by making them parity preserving. The importance of this proposed work lies in the fact that it provides the design of reversible sequential circuits completely testable for any stuck-at fault and single bit fault. In our opinion our design of reversible building blocks is superior to existing designs in term of quantum cost, hardware complexity, constant input, garbage output, number of gates and design of online testable D flip-flop have been proposed for the first time. We hope our work can be extended for building complex reversible sequential circuits.

Keywords: parity preserving gate, quantum computing, fault tolerance, flip-flop, sequential reversible logic

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Authors: Tong Ming Liu

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Human mesenchymal stem cells (MSCs) represent the most used stem cells for clinical application, which have been used in over 1000 clinical trials to treat over 30 diseases due to multilineage differentiation potential, secretome and immunosuppression. Gene therapies of MSCs hold great promise in the treatment of many diseases due to enhanced MSC-based clinical outcomes. To identify genes for gene therapy of MSCs, by comparing gene expression profile before and after MSC differentiation following by functional screening, we have identified ZNF145 that regulated MSC differentiation. Forced expression of ZNF145 resulted in enhanced in vitro chondrogenesis of MSCs as an upstream factor of SOX9 and improved osteochondral repair upon implant into osteochondral defects in rodents. By comparing gene expression profile during differentiation of iPSCs toward MSCs, we also identified gene HOX regulating MSC stemness, which was much downregulated in late-passaged MSCs. Knockdown of this gene greatly compromised MSC stemness including abolished proliferation, decreased CFU-F, promoted senescence and reduced expression of cell surface antigens linked to the MSC phenotype. In addition, multi-linage differentiation was also greatly impaired. Notably, HOX overexpression resulted in improved multi-lineage differentiation. In the mechanism, HOX expression significantly deceased in late passage of MSCs compared with early passage of MSCs, correlating with MSC important genes. ChIP-seq data shown that HOX binds to genes related to MSC self-renewal and differentiation. Most importantly, most HOX binding sites are lost in late passage of MSCs. HOX exerts its effects by directing binding Twist1, one important gene of MSCs. The identification of the genes regulating MSC differentiation and stemness will provide and promising strategies for gene therapy of MSCs in regenerative medicine.

Keywords: mesenchymal stem cell, novel transcription factor, stemness, gene therapy, cartilage repair, signaling pathway

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Authors: Maryam S. Ghoraishi, John E. Hawk, Thomas Thundat

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Understanding liquid properties in small scale has become important in recent decades as immerging new microelectromechanical systems (MEMS) devices have been widely used for micro pumps, drug delivery, and many other laboratory-on-microchips analysis. Often in microfluidic devices, fluids are transported electrokinetically. Therefore, extensive knowledge of fluid flow, heat transport, electrokinetics and electrochemistry are key to successful lab on a chip design. Among different microfluidic devices, recently developed hollow channel cantilever offers an ideal platform to study different fluid properties simultaneously without drastic decrease in quality factor which normally occurs when traditional cantilevers operate in the liquid phase. Using hollow channel cantilever, we monitor changes in density and viscosity of liquid while simultaneously investigating dielectric properties of alcohol water binary mixtures. Considerable research has been conducted on alcohol-water mixtures since such a mixture is a typical prototype for biomolecules, Micelle formation, and structural stability of proteins (to name a few). Here we show that hollow channel cantilever can be employed to investigate dielectric properties of ethanol/water mixtures in different concentrations. We study dynamic amplitude shifts of hollow channel cantilever oscillation at different concentrations of ethanol/water for different voltages. Our results show how interactions between solute and solvent, and possibly cluster formation, could change dielectric properties and dipole reorientation of the mixture, as well as the resulting force on the hollow cantilever. For comparison, we also examine higher conductivity ionic mixtures of sodium sulfate solution under the same conditions as low conductivity ethanol/water mixtures. We will show the results from systematic investigation of solvent effects on dielectric properties of the binary mixture. We will also address the question of resolution limits in dielectric study of analyte molecules imposed by solvent concentrations.

Keywords: dielectric constant, cantilever sensors, ethanol water mixtures, low frequency

Procedia PDF Downloads 175

Authors: Abbas Ali Husseini, Ali Mohammad Yazdani, Fatemeh Ghadiri, Alper Şişman

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We developed a microchip device that uses surface acoustic waves for rapid lysis of low level of cell samples. The device incorporates sharp-edge glass microparticles for improved performance. We optimized the lysis conditions for high efficiency and evaluated the device's feasibility for point-of-care applications. The microchip contains a 13-finger pair interdigital transducer with a 30-degree focused angle. It generates high-intensity acoustic beams that converge 6 mm away. The microchip operates at a frequency of 16 MHz, exciting Rayleigh waves with a 250 µm wavelength on the LiNbO3 substrate. Cell lysis occurs when Candida albicans cells and glass particles are placed within the focal area. The high-intensity surface acoustic waves induce centrifugal forces on the cells and glass particles, resulting in cell lysis through lateral forces from the sharp-edge glass particles. We conducted 42 pilot cell lysis experiments to optimize the surface acoustic wave-induced streaming. We varied electrical power, droplet volume, glass particle size, concentration, and lysis time. A regression machine-learning model determined the impact of each parameter on lysis efficiency. Based on these findings, we predicted optimal conditions: electrical signal of 2.5 W, sample volume of 20 µl, glass particle size below 10 µm, concentration of 0.2 µg, and a 5-minute lysis period. Downstream analysis successfully amplified a DNA target fragment directly from the lysate. The study presents an efficient microchip-based cell lysis method employing acoustic streaming and microparticle collisions within microdroplets. Integration of a surface acoustic wave-based lysis chip with an isothermal amplification method enables swift point-of-care applications.

Keywords: cell lysis, surface acoustic wave, micro-glass particle, droplet

Procedia PDF Downloads 52

Authors: Andrés Quiroz, Emilio Hormazabal, Ana Mutis, Fernando Ortega, Manuel Chacón-Fuentes, Leonardo Parra

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Red clover (Trifolium pratense L.) is an important forage species in temperate regions of the world. The main limitation of this species worldwide is a lack of persistence related to the high mortality of plants due to a complex of biotic and abiotic factors, determining a life span of two or three seasons. Because of the importance of red clover in Chile, a red clover breeding program was started at INIA Carillanca Research Center in 1989, with the main objective of improving the survival of plants, forage yield, and persistence. The main selection criteria for selecting new varieties have been based on agronomical parameters and biotic factors. The main biotic factor associated with red clover mortality in Chile is Hylastinus obscurus (Coleoptera: Curculionidae). Both larval and adults feed on the roots, causing weakening and subsequent death of clover plants. Pesticides have not been successful for controlling infestations of this root borer. Therefore, alternative strategies for controlling this pest are a high priority for red clover producers. Currently, the role of semiochemical in the interaction between H. obscurus and red clover plants has been widely studied for our group. Specifically, from the red clover foliage has been identified limonene is eliciting repellency from the root borer. Limonene is generated in the plant from two independent biosynthetic pathways, the mevalonic acid, and deoxyxylulose pathway. Mevalonate pathway enzymes are localized in the cytosol, whereas the deoxyxylulose phosphate pathway enzymes are found in plastids. In summary, limonene can be determinated by enzymatic bioassay using GPP as substrate and by limonene synthase expression. Therefore, the main objective of this work was to study genetic variation of limonene in material provided by INIA´s Red Clover breeding program. Protein extraction was carried out homogenizing 250 mg of leave tissue and suspended in 6 mL of extraction buffer (PEG 1500, PVP-30, 20 mM MgCl2 and antioxidants) and stirred on ice for 20 min. After centrifugation, aliquots of 2.5 mL were desalted on PD-10 columns, resulting in a final volume of 3.5 mL. Protein determination was performed according to Bradford with BSA as a standard. Monoterpene synthase assays were performed with 50 µL of protein extracts transferred into gas-tight 2 mL crimp seal vials after addition of 4 µL MgCl₂ and 41 µL assay buffer. The assay was started by adding 5 µL of a GPP solution. The mixture was incubated for 30 min at 40 °C. Biosynthesized limonene was quantified in a GC equipped with a chiral column and using synthetic R and S-limonene standards. The enzymatic the production of R and S-limonene from different Superqueli-Carillanca genotypes is shown in this work. Preliminary results showed significant differences in limonene content among the genotypes analyzed. These results constitute an important base for selecting genotypes with a high content of this repellent monoterpene towards H. obscurus.

Keywords: head space, limonene enzymatic determination, red clover, Hylastinus obscurus

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